III. Unidades de Investigação, Desenvolvimento e Outras
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- 1,2,4-Trioxolane and 1,2,4,5-Tetraoxane endoperoxides against old-world Leishmania parasites: in vitro activity and mode of actionPublication . Mendes, Andreia; Armada, Ana; Cabral, Lília; Amado, Patrícia; Campino, Lenea; Cristiano, Maria de Lurdes; Cortes, SofiaLeishmaniasis remains one of the ten Neglected Tropical Diseases with significant morbidity and mortality in humans. Current treatment of visceral leishmaniasis is difficult due to a lack of effective, non-toxic, and non-extensive medications. This study aimed to evaluate the selectivity of 12 synthetic endoperoxides (1,2,4-trioxolanes; 1,2,4,5-tetraoxanes) and uncover their biochemical effects on Leishmania parasites responsible for visceral leishmaniasis. The compounds were screened for in vitro activity against L. infantum and L. donovani and for cytotoxicity in two monocytic cell lines (J774A.1 and THP-1) using the methyl thiazol tetrazolium assay. Reactive oxygen species formation, apoptosis, and mitochondrial impairment were measured by flow cytometry. The compounds exhibited fair to moderate anti-proliferative activity against promastigotes of the 2 Leishmania species, with IC50 values ranging from 13.0 ± 1.7 µM to 793.0 ± 37.2 µM. Tetraoxanes LC132 and LC138 demonstrated good leishmanicidal activity on L. infantum amastigotes (IC50 13.2 ± 5.2 and 23.9 ± 2.7 µM) with low cytotoxicity in mammalian cells (SIs 22.1 and 118.6), indicating selectivity towards the parasite. Furthermore, LC138 was able to induce late apoptosis and dose-dependent oxidative stress without affecting mithocondria. Compounds LC132 and LC138 can be further explored as potential antileishmanial chemotypes.
- 1-Methylcyclopropene and lemongrass essential oil nanocoatings effect on the preservation of cold stored ‘Rocha’ pearPublication . Gago, Custódia; Guerreiro, Adriana; Cruz, Sandra; Martins, Nuno; Cabrita, Maria João; Miguel, Maria; Faleiro, Maria Leonor; Antunes, Maria DulceThe effects of coating 'Rocha' pear with alginate-based nanoemulsions enriched with lemongrass essential oil (LG) was evaluated and compared to the usual 1-MCP treatment. Fruit were treated with 1-MCP (312 nL L-1) or coated with nanoemulsions: sodium alginate 2 % (w/w) + lemongrass essential oil 1.25 % (w/w) (LG 1.25 %) or lemongrass essential oil 2.5 % (w/w) (LG 2.5 %). Then, fruit were stored at 0 degrees C and 90-95 % relative humidity (RH), for eight months. Fruit samples were collected at harvest and after two, four, six and eight months of cold storage, and then transferred to shelf-life at 22 degrees C. Upon removal and after 7 d shelf-life, fruit symptoms of superficial scald and internal browning, ethylene production, color CIE (L*, hue), firmness, soluble solids content (SSC), titratable acidity (TA), weight loss, electrolytic leakage (EL), antioxidant activity and fatty acids of pear peel, microbial growth and sensory analyses were evaluated. Coatings and 1-MCP reduced fruit color evolution and preserved better firmness than control. Coatings and 1-MCP did not affect SSC and TA. Treatments did not influence the sensory quality. Microbial growth was within the safety limits in all treatments. Treatments with 1-MCP and LG-nanoemulsions were similarly efficient to reduce superficial scald, nevertheless the LG-nanoemulsions showed higher internal disorders after 8 months of storage and LG 2.5 % had higher decay at the same period, similar to control. 1-MCP treated fruit had the lowest softening rate after shelf-life up to 4 months and LG 2.5 % showed higher weight loss. Also, ethylene production was higher in control and LG 1.25 % up to 6 months plus shelf-life, while after 8 months there was no difference among treatments. This study suggests that 1-MCP is the most efficient for preserving quality of 'Rocha' pear for 8 months, while up to 6 months the best effect is obtained with LG 1.25 % nanocoatings.
- A 1000-year-old case of Klinefelter's syndrome diagnosed by integrating morphology, osteology, and geneticsPublication . Roca-Rada, Xavier; Tereso, Sofia; Rohrlach, Adam B.; Brito, Andre; Williams, Matthew P.; Umbelino, Cláudia; Curate, Francisco; Deveson, Ira W.; Souilmi, Yassine; Amorim, Antonio; Carvalho, Pedro C.; Llamas, Bastien; Teixeira, Joao C.The skeleton of a human being buried over 1000 years ago, in the medieval archaeological site of Torre Velha, in north-eastern Portugal, was referred to us for analysis and investigation
- 11-ketotestosterone inhibits the alternative mating tactic in sneaker males of the peacock blenny, Salaria pavoPublication . Oliveira, R. F.; Carneiro, L. A.; Gonçalves, D.; Canario, Adelino; Grober, M. S.In the peacock blenny, Salaria pavo, a species with courtship sex-role reversal, smaller, younger males mimic the courtship behavior and the nuptial coloration of females in order to get access to nests during spawning and to parasitize egg fertilization from nest-holder males. Later in their life, sneakers transform both morphologically and behaviorally into nest-holder males. In the present paper we investigate the activational role of 11-ketotestosterone (KT), the most potent androgen in most teleost species, to promote the switch between tactics in sneaker males of S. pavo. Sneakers were implanted either with KT or with control (i.e. castor oil) silastic implants. A week after implantation they were subjected to a set of behavioral tests and morphometric measurements. KT treatment promoted the differentiation of secondary sex characters, such as the anal glands, and inhibited the expression of female courtship behavior. KT-treated sneakers also showed a trend toward less frequent display of female nuptial coloration. There was no effect of KT treatment on the expression of typical nest-holder male behavior. Finally, there was no effect of KT treatment on the number or soma size of arginine vasotocin neurons in the preoptic area, which are often associated with the expression of vertebrate sexual behavior. Thus, KT seems to play a key role in mating tactic switching by inhibiting the expression of female courtship behavior and by promoting the development of male displaying traits (e.g. anal glands). The lack of a KT effect on behavior typical of nest-holding males and vasotocinergic preoptic neurons suggests that a longer time frame or other endocrine/social signals are needed for the initiation of these traits in males that are switching tactics.
- 11-ketotestosterone stimulates putative sex pheromone production in the male peacock blenny, Salaria pavo (Risso 1810)Publication . Serrano, Rui M.; Lopes, Orlando; Hubbard, Peter; Araújo, J.; Canario, Adelino V. M.; Barata, E. N.Male peacock blennies (Salaria pavo) release odors from their anal glands and, possibly, from their gonads that attract reproductive females. The current study investigated the effects of 11-ketotestosterone (KT) on development of the anal glands, testes, and accessory testicular organs as well as the subsequent olfactory potency of their secretions. After 3 wk of KT treatment (5 mg/kg in silastic implants), clusters of cells secreting neutral mucins differentiated in the anal gland of all treated males, whereas this occurred in only one control male. Secretions by anal glands from KT-treated males elicited greater olfactory responses, as assessed by electro-olfactography, than those from controls. Treatment with KT stimulated testicular gland growth and sialomucin secretion but had no clear effect on the germinal region of the testis; KT also stimulated enlargement of, and fluid secretion in, the blind pouches (paired evaginations of the spermatic ducts). Secretions by the testes and fluid in blind pouches from KT-treated males elicited greater olfactory responses than those from controls. In conclusion, KT stimulates development of the anal glands and testicular accessory organs and promotes odorant secretion, the putative multicomponent male peacock blenny pheromone.
- A 12,000-yr pollen record off Cape Hatteras — Pollen sources and mechanisms of pollen dispersionPublication . Naughton, F.; Keigwin, L.; Peteet, D.; Costas, S.; Desprat, S.; Oliveira, Dulce; de Vernal, A.; H L Voelker, Antje; Abrantes, FatimaIntegrating both marine and terrestrial signals from the same sediment core is one of the primary challenges for understanding the role of ocean-atmosphere coupling throughout past climate changes. It is therefore vital to understand how the pollen signal of a given marine record reflects the vegetation changes of the neighboring continent. The comparison between the pollen record of marine core JPC32 (KNR178JPC32) and available terrestrial pollen sequences from eastern North America over the last 12,170 years indicates that the pollen signature off Cape Hatteras gives an integrated image of the regional vegetation encompassing the Pee Dee river, Chesapeake and Delaware hydrographic basins and is reliable in reconstructing the past climate of the adjacent continent. Extremely high quantities of pollen grains included in the marine sediments off Cape Hatteras were transferred from the continent to the sea, at intervals 10,100-8800 cal yr BP, 8300-7500 cal yr BP, 5800-4300 cal yr BP and 2100-730 cal yr BP, during storm events favored by episodes of rapid sea-level rise in the eastern coast of US. In contrast, pollen grains export was reduced during 12,170-10,150 cal yr BP and 4200-2200 cal yr BP, during episodes of intense continental dryness and slow sea level rise episodes or lowstands in the eastern coast of US. The near absence of reworked pollen grains in core JPC32 contrasts with the high quantity of reworked material in nearby but deeper located marine sites, suggesting that the JPC32 record was not affected by the Deep Western Boundary Current (DWBC) since the end of the Younger Dryas and should be considered a key site for studying past climate changes in the western North Atlantic.
- A 15-month survey of Dimethylsulfoniopropionate and Dimethylsulfoxide content in Posidonia oceanicaPublication . Richir, Jonathan; Champenois, Willy; Engels, Guyliann; Abadie, Arnaud; Gobert, Sylvie; Lepoint, Gilles; Silva, João; Santos, Rui; Sirjacobs, Damien; V. Borges, AlbertoPosidonia oceanica is the only reported seagrass to produce significant amount of dimethylsulfoniopropionate (DMSP). It is also the largest known producer of DMSP among coastal and inter-tidal higher plants. Here, we studied (i) the weekly to seasonal variability and the depth variability of DMSP and its related compound dimethylsulfoxide (DMSO) in P. oceanica leaves of a non-disturbed meadow in Corsica, France, (ii) the weekly to seasonal variability and the depth variability of DMSP to DMSO concentration to assess the potential of the DMSP:DMSO ratio as indicator of stress, and (iii) the relationships between DMSP, DMSO, and the DMSP:DMSO ratio with potential explanatory variables such as light, temperature, photosynthetic activity (effective quantum yield of photosystem II), and leaf size. The overall average concentrations of organosulfured compounds in P. oceanica leaves were 130 ± 39 µmol.g−1 fw for DMSP and 4.9 ± 2.1 µmol.g−1 fw for DMSO. Concentrations of DMSP and DMSO in P. oceanica were overall distinctly higher and exhibited a wider range of variations than other marine primary producers such as Spartina alterniflora, phytoplankton communities, epilithic Cyanobacteria and macroalgae. Concentrations of both DMSP and DMSO in P. oceanica leaves decreased from a maximum in autumn to a minimum in summer; they changed little with depth. Potential explanatory variables except the leaf size, i.e., the leaf age were little or not related to measured concentrations. To explain the seasonal pattern of decreasing concentrations with leaf aging, we hypothesized two putative protection functions of DMSP in young leaves: antioxidant against reactive oxygen species and predator-deterrent. The similar variation of the two molecule concentrations over time and with depth suggested that DMSO content in P. oceanica leaves results from oxidation of DMSP. The DMSP:DMSO ratio remained constant around a mean value of 29.2 ± 9.0 µmol:µmol for the non-disturbed harvested meadow regardless of the time of the year, the depth or the leaf size. As suggested for the salt march plant S. alterniflora, we hypothesized the DMSP:DMSO ratio could be considered as indicator of stress in seagrasses exposed to environmental or anthropogenic stressors. More research would now be needed to confirm the functions of DMSP and DMSO in seagrasses and how the DMSP:DMSO ratio will vary under various disturbances.
- 16.P15. Branchial osmoregulatory response to salinity challenge in the Lusitanian toadfishPublication . Guerreiro, Pedro M; Laiz-Carrión, R.; Haond, Christophe; Modesto, Teresa; Fuentes, Juan; Mancera, J. M.; Canario, Adelino V. M.Halobatrachus didactylus, a marine teleost found in coastal lagoons and river estuaries is often exposed to important salinity changes. Despite its aglomerular kidney, it is able to survive in hypo-osmotic environments, likely via compensatory actions from gills and intestine. We aimed at evaluating the response of the branchial tissue of H. didactylus to salinity changes.
- 17a,20ß,21-trihydroxy-4-pregnen-3-one: the probable maturation-inducing steroid of the lusitanian toadfishPublication . Modesto, Teresa; Canario, Adelino V. M.17,20 ,21-Trihydroxy-4-pregnen-3-one (17,20 ,21-P) was identified as the major metabolite of incubations of Lusitanian toadfish Halobatrachus didactylus ovarian follicles with [3H]-17hydroxyprogesterone. The potency of several steroids in inducing germinal vesicle breakdown of follicle-enclosed oocytes of Lusitanian toadfish was systematically examined by using an in vitro germinal vesicle breakdown (GVBD) bioassay. 17,20 -Dihydroxy-4-pregnen-3-one (17,20 -P) and 17,20 ,21-P, two confirmed maturation-inducing steroids (MIS) in teleosts, were the most potent in inducing GVBD with ED50s ranging between 9 and 271 nM. Structure-activity relationships followed similar patterns to what has been observed in similar bioassays, i.e. a vital requirement for 17- and 20 -hydroxyl groups in C21 steroids and a reduction in activity of 14 and 5–6%, respectively, for 5-pregnene and 5 -pregnanes compared to 4-pregnenes. Corticosteroids, testosterone and 17 -oestradiol were ineffective. Folliculated oocytes stimulated by pituitary homogenate produced 17,20 ,21-P from endogenous substrates in amounts one order of magnitude higher than 17,20 -P. These results strongly support the hypothesis that 17,20 ,21-P is the likely MIS in this species.
- 17α-ethynylestradiol prevents the natural male-to-female sex change in gilthead seabream (Sparus aurata L.)Publication . García Hernández, M. Pilar; Cabas, Isabel; Rodenas, M. Carmen; Arizcun, Marta; Chaves-Pozo, Elena; Power, Deborah; García Ayala, AlfonsaExposure to 17α-ethynylestradiol (EE2, 5 μg/g food) impairs some reproductive events in the protandrous gilthead seabream and a short recovery period does not allow full recovery. In this study, spermiating seabream males in the second reproductive cycle (RC) were fed a diet containing 5 or 2.5 μg EE2/g food for 28 days and then a commercial diet without EE2 for the remaining RC. Individuals were sampled at the end of the EE2 treatment and then at the end of the RC and at the beginning of the third RC, 146 and 333 days after the cessation of treatment, respectively. Increased hepatic transcript levels of the gene coding for vitellogenin (vtg) and plasma levels of Vtg indicated both concentrations of EE2 caused endocrine disruption. Modifications in the histological organization of the testis, germ cell proliferation, plasma levels of the sex steroids and pituitary expression levels of the genes coding for the gonadotropin β-subunits, fshβ and lhβ were detected. The plasma levels of Vtg and most of the reproductive parameters were restored 146 days after treatments. However, although 50% of the control fish underwent sex reversal as expected at the third RC, male-to female sex change was prevented by both EE2 concentrations.