Rapamycin reverses age-related increases in mitochondrial ROS production at complex I, oxidative stress, accumulation of mtDNA fragments inside nuclear DNA, and lipofuscin level, and increases autophagy, in the liver of middle-aged mice

Rapamycin consistently increases longevity in mice although the mechanism of action of this drug is unknown. In the present investigation we studied the effect of rapamycin on mitochondrial oxidative stress at the same dose that is known to increase longevity in mice (14 mg of rapamycin/kg of diet). Middle aged mice (16 months old) showed significant age-related increases in mitochondrial ROS production at complex I, accumulation of mtDNA fragments inside nuclear DNA, mitochondrial protein lipoxidation, and lipofuscin accumulation compared to young animals (4 months old) in the liver. After 7 weeks of dietary treatment all those increases were totally or partially (lipofuscin) abolished by rapamycin, middle aged rapamycin-treated animals showing similar levels in those parameters to young animals. The decrease in mitochondrial ROS production was due to qualitative instead of quantitative changes in complex I. The decrease in mitochondrial protein lipoxidation was not due to decreases in the amount of highly oxidizable unsaturated fatty acids. Rapamycin also decreased the amount of RAPTOR (of mTOR complex) and increased the amounts of the PGC1-α and ATG13 proteins. The results are consistent with the possibility that rapamycin increases longevity in mice at least in part by lowering mitochondrial ROS production and increasing autophagy, decreasing the derived final forms of damage accumulated with age which are responsible for increased longevity. The decrease in lipofuscin accumulation induced by rapamycin adds to previous information suggesting that the increase in longevity induced by this drug can be due to a decrease in the rate of aging. © 2016 Elsevier Inc.

Age and growth of the undulate ray, Raja undulata, in the Algarve (southern Portugal)

Aspects of the population dynamics of the undulate ray, Raja undulata (Chondrichthyes: Rajidae), one of the more abundant elasmobranch fish captured along the Portuguese south coast (Algarve), were studied for the first time. Some traditional elasmobranch vertebral enhancing techniques were compared and the most precise for this species determined to be the cedar wood oil immersion and the alizarin red S stain. The sample consisted of 14 age-classes, with age-classes 3 to 8 being the most represented. Evidence of an annual deposition pattern of a pair of bands (one opaque and one translucent) was found by marginal increment analyses. Von Bertalanffy growth parameters were estimated and no differences found between males and females (all data: L-inf=110.22 cm, K=0.11 y(-1) and t(0)=-1.58 y).

Ecological quality assessment of transitional waters based on fish assemblages in Portuguese estuaries: The Estuarine Fish Assessment Index (EFAI)

The assessment of water quality has changed markedly worldwide over the last years, especially in Europe due to the implementation of the Water Framework Directive. Fish was considered a key-element in this context and several fish-based multi-metric indices have been proposed. In this study, we propose a multi-metric index, the Estuarine Fish Assessment Index (EFAI), developed for Portuguese estuaries, designed for the overall assessment of transitional waters, which could also be applied at the water body level within an estuary. The EFAI integrates seven metrics: species richness, percentage of marine migrants, number of species and abundance of estuarine resident species, number of species and abundance of piscivorous species, status of diadromous species, status of introduced species and status of disturbance sensitive species. Fish sampling surveys were conducted in 2006, 2009 and 2010, using beam trawl, in 13 estuarine systems along the Portuguese coast. Most of the metrics presented a high variability among the transitional systems surveyed. According to the EFAI values, Portuguese estuaries presented a "Good" water quality status (except the Douro in a particular year). The assessments in different years were generally concordant, with a few exceptions. The relationship between the EFAI and the Anthropogenic Pressure Index (API) was not significant, but a negative and significant correlation was registered between the EFAI and the expert judgement pressure index, at both estuary and water body level. The ordination analysis performed to evaluate similarities among North-East Atlantic Geographical Intercalibration Group (NEAGIG) fish-based indices put in evidence four main groups: the French index, since it is substantially different from all the other indices (uses only four metrics based on densities); indices from Ireland, United Kingdom and Spain (Asturias and Cantabria); the Dutch and German indices; and the indices of Belgium. Portugal and Spain (Basque country). The need for detailed studies, including comparative approaches, on several aspects of these assessment tools, especially in what regards their response to anthropogenic pressures was stressed. (C) 2011 Elsevier Ltd. All rights reserved.

Reproductive aspects of the undulate ray, Raja undulata, from the south coast of Portugal

The maturity of the undulate ray, Raja undulata, one of the most important by-catch elasmobranch species captured in south Portugal, was studied for the first time. Females ranged in total length (TL) from 19.4 to 88.2 cm and males from 23.0 to 83.2 cm. The gonadosomatic index for females was higher during the winter, meaning that this species reproduces during this season. Females matured at significantly larger sizes and older ages than males, with 50% of the females mature at 76.2 cm TL at an age of 8.98 years while 50% maturity for males was achieved at 73.6 cm TL and 7.66 years. Females were mature at 86.3% of the maximum observed size and 69.1% of the maximum observed age while males matured at 88.5% of the maximum observed size and 63.8% of the maximum observed age, making this a very late maturing species, with important consequences in terms of fisheries management and conservation. (c) 2006 Elsevier B.V. All rights reserved.

European lampreys: new insights on postglacial colonization, gene flow and speciation.

Ice ages are known to be the most dominant palaeoclimatic feature occurring on Earth, producing severe climatic oscillations and consequently shaping the distribution and the population structure of several species. Lampreys constitute excellent models to study the colonization of freshwater systems, as they commonly appear in pairs of closely related species of anadromous versus freshwater resident adults, thus having the ability to colonize new habitats, through the anadromous species, and establish freshwater resident derivates. We used 10 microsatellite loci to investigate the spatial structure, patterns of gene flow and migration routes of Lampetra populations in Europe. We sampled 11 populations including the migratory L. fluviatilis and four resident species, L. planeri, L. alavariensis, L. auremensis and L. lusitanica, the last three endemic to the Iberian Peninsula. In this southern glacial refugium almost all sampled populations represent a distinct genetic cluster, showing high levels of allopatric differentiation, reflecting long periods of isolation. As result of their more recent common ancestor, populations from northern Europe are less divergent among them, they are represented by fewer genetic clusters, and there is evidence of strong recent gene flow among populations. These previously glaciated areas from northern Europe may have been colonized from lampreys expanding out of the Iberian refugia. The pair L. fluviatilis/L. planeri is apparently at different stages of speciation in different locations, showing evidences of high reproductive isolation in the southern refugium, and low differentiation in the north.

Development and application of a selenium speciation method in cattle feed and beef samples using HPLC-ICP-MS: evaluating the selenium metabolic process in cattle.

2016

Fibrinolysis Versus Primary Pci In Stemi Patients Enrolled In The International Survey Of Acute Coronary Syndromes In Transitional Countries

Primary angioplasty has been shown to be more effective than fibrinolysis in terms of mortality and adverse outcomes. More recent data, however, suggests that timely reperfusion with fibrinolysis is comparable to primary angioplasty. The current study gathered data from the International Survey of Acute Coronary Syndromes in Transitional Countries registry. Among 7406 ST-elevation myocardial infarction patients presenting within 12 hours from symptom onset, 6315 underwent primary percutaneous coronary intervention and 1091 were treated with fibrinolysis. The primary outcome was 30-day mortality, while the secondary outcome was a composite of 30-day incidence of death, severe left ventricular dysfunction, stroke or reinfarction. Patients who underwent primary angioplasty tended to have a greater cardiovascular risk profile and were slightly older. On the other hand, patients treated with fibrinolysis received less anti-platelet medications yet were more often prescribed beta blockers in the acute phase. Among those who received fibrinolysis, 43% underwent coronary angiography while 32.3% were treated with a subsequent angioplasty. Total ischemic time was lower in patients undergoing fibrinolysis (185 minutes) than in those treated with primary angioplasty (258 minutes). Rates of primary and secondary combined endpoints were higher in patients receiving fibrinolysis compared to those receiving primary angioplasty (7.8% vs. 4.1%; p<0.0001; OR 1.97, 95% CI, 1.38-2.81; and 14.8% vs. 10.1%, p<0.0001; OR 1.43, 95% CI, 1.12-1.81). When considering only patients receiving reperfusion within 3 hours, regardless of reperfusion strategy, differences in mortality (6.3% vs. 4%, p=0.094, for fibrinolysis or primary angioplasty, respectively; OR 0.87, 95% CI, 0.35-2.16) and in the combined secondary endpoint were no longer observed (12.9% vs 10.8%, p=0.33; OR 0.98, 95% CI, 0.58-1.64), and female sex was no longer a significant predictor of adverse outcomes. When performed 3 hours from symptom onset, fibrinolysis is safe and feasible, in terms of mortality and adverse outcomes, compared to primary angioplasty.

Effects of different hypoxia timing regimes on a transitional habitat's ( Pialassa Baiona, Ravenna, Italy) benthic community.

The aim of this thesis was to quantify experimentally in the field the effects of different timing regimes of hypoxia on the structure of benthic communities in a transitional habitat. The experiment was performed from 8 July to 29 July 2019 in a shallow subtidal area in Pialassa Baiona (Italy), a lagoon characterized by mixing regimes dominated by the tide. The benthic community was isolated using cylinders 15,5Cm x 20Cm size. Hypoxic conditions were imposed by covering the treated cylinders with a black plastic bag while control cylinders were left uncovered. We created 4 different timing regimes of hypoxia by manipulating both the duration of hypoxia (4 or 8 days) as well as the ratio between the duration of subsequent periods of hypoxia and the duration of a normoxic period between subsequent hypoxic events (D4R3/2, D8R3/2). At the end of each experimental trial, the benthic communities within each pot were retrieved, sieved in the field and subsequent analyzed in the laboratory where organisms were identified and counted. Results showed that benthic organism were generally negatively affected by hypoxic stress events. As expected, longer hypoxic events caused a stronger decrease of benthic community abundance. When the hypoxic events were interrupted by the normoxic event there were two different results. If the hypoxic period was too long, the normoxic period didn’t cause a positive recovery effect, and further decline of the benthic community was observed. Conversely normoxia had positive effects if the period of hypoxia was short enough not to compromise the benthic community. This resulted in a statistically significant interaction between the tested factors Duration and Ratio. Amphipods were the most sensitive organisms to hypoxia. We conclude that the effects of hypoxia can be greatly relieved by short normoxic periods if they happen frequently enough.

Effects of stochastic hypoxia events on the benthic communities of transitional waters

Hypoxia is one of the most important and faster spreading threats to marine life and its occurrence has significantly increased in the last century. The effects of hypoxia on marine organisms and communities has mostly been studied in light of the intensity of the disturbance but not a lot of attention has been given to its interaction with other stressors and the timing of its appearance. In this thesis I started to explore these topics through laboratory and manipulative field experiments. I studied the interactive effects of thermal stress and hypoxia on a European native bivalve species (Cerastoderma edule; Linnaeus, 1758 ) and a non native one (Ruditapes philippinarum; Adams & Reeve, 1850) through a laboratory experiment performed in the Netherlands. The non native species displayed a greater tolerance to oxygen depletion than the native one. The first field experiment was performed in an Italian brackish coastal lagoon (Pialassa Baiona) and tested the effects of different timing regimes of hypoxia on the benthic community. It emerged that the main factor affecting the community is the duration of the hypoxia. The ability of the communities to recover after repeated hypoxic periods was explored in the second manipulative field experiment. We imposed three different timing regimes of hypoxia on sediment patches in Pialassa Baiona and we monitored the changes of both the benthic and the microbial communities after the disturbances. The preliminary analyses of the data from this last work suggest that the experimental manipulations caused limited detrimental effects on the communities. Overall this thesis work suggests that the duration of hypoxic events, their repetitive nature and the associated thermal stress are key factors in determining their effects on the communities and that management measures should point towards a reduction of the duration of the single hypoxic periods more than their frequency.

Transitional Justice as a Post-Revolution Constitutional Arrangement: A Law and Economics Approach

This dissertation addresses the timely questions of transitional justice (TJ) in the aftermath of revolutions against autocratic regimes, dealing with TJ as a constitutional arrangement through the lenses of constitutional economics. After an introductory chapter, chapter 2 deals with why nations rarely adopt meaningful TJ processes in the first place, it then explains the limitations of civil society as the arbiter, facilitator, and enforcer of TJ policies. Chapter 3 tackles the question of which mechanisms to choose? It uses the UN Guidelines on TJ that sets five principal TJ mechanisms. It provides a cost-benefit analysis (CBA) of each mechanism and suggests policy implications accordingly. The CBA inspires chapter 4 analysis, suggesting a tradeoff between restrictive fair trial standards under constitutional laws and justice considerations. The tradeoff explains the suggested efficiency of the balanced TJ approaches that combine trials and amnesties. This approach is used for the case study analysis of TJ in Tunisia after the 2011 revolution in chapter 5. The chapter presents the first index of TJ mechanisms in Tunisia through novel data collected by the author. It shows an ultimate TJ design that ended with a modest harvest in the application. The lack of cooperation between the Tunisian parties, added to the absence of transparency in many TJ measures, threatens any possible positive outcomes of the partial TJ process. It is also alarming regarding constitutional compliance in a system that – until recently - was considered the only democracy in the Arab region. Chapter 6 is a summary

Ghost Cells In Pilomatrixoma, Craniopharyngioma, And Calcifying Cystic Odontogenic Tumor: Histological, Immunohistochemical, And Ultrastructural Study.

Pilomatrixoma, craniopharyngioma, and calcifying cystic odontogenic tumor are the main entities presenting ghost cells as an important histological feature, in spite their quite different clinical presentation; it seems that they share a common pathway in the formation of these cells. The aim of this study is to examine and compare the characteristics of ghost and other cells that form these lesions. Forty-three cases including 21 pilomatrixomas, 14 craniopharyngiomas, and eight calcifying cystic odontogenic tumors were evaluated by immunohistochemistry for cytokeratins, CD138, β-catenin, D2-40, Glut-1, FAS, CD10 and also by scanning electron microscopy. The CKs, CD138, β-catenin, Glut-1, FAS, and CD10 were more often expressed by transitional cells of craniopharyngioma and calcifying cystic odontogenic tumor, compared with pilomatrixoma. Basaloid cells of pilomatrixoma showed strong positivity for CD138 and CD10. Differences on expression pattern were identified in transitional and basal cells, as ghost cells were negative for most antibodies used, except by low expression for cytokeratins. By scanning electron microscopy, the morphology of ghost cells were similar in their fibrillar cytoplasm, but their pattern varied from sheets in pilomatrixoma to small clusters in craniopharyngioma and calcifying cystic odontogenic tumor. Mechanisms involved in formation of ghost cells are unknown, but probably they follow different pathways as protein expression in the basal/transitional cells was not uniform in the three tumors studied.

Transition From Glass To Digital Slide Microscopy In The Teaching Of Oral Pathology In A Brazilian Dental School.

Several medical and dental schools have described their experience in the transition from conventional to digital microscopy in the teaching of general pathology and histology disciplines; however, this transitional process has scarcely been reported in the teaching of oral pathology. Therefore, the objective of the current study is to report the transition from conventional glass slide to virtual microscopy in oral pathology teaching, a unique experience in Latin America. An Aperio ScanScope® scanner was used to digitalize histological slides used in practical lectures of oral pathology. The challenges and benefits observed by the group of Professors from the Piracicaba Dental School (Brazil) are described and a questionnaire to evaluate the students' compliance to this new methodology was applied. An improvement in the classes was described by the Professors who mainly dealt with questions related to pathological changes instead of technical problems; also, a higher interaction with the students was described. The simplicity of the software used and the high quality of the virtual slides, requiring a smaller time to identify microscopic structures, were considered important for a better teaching process. Virtual microscopy used to teach oral pathology represents a useful educational methodology, with an excellent compliance of the dental students.

Estudo teórico da interação existente entre a artemisinina e o heme

The purpose of this work is to study theoretically stereoelectronic aspects of the interaction between heme and artemisinin in the transitional heme-artemisinin complex. Through semi-empirical calculations using the PM3 method, the potential energy barrier of artemisinin rotation relative to heme in the heme-artemisinin complex was studied in vacuum and in the partially solvated state. The minimum heat of formation obtained for the complex with and without water molecules is -702.39 and -100.86 kcal mol-1, respectively, which corresponds to the dihedral angle C-Fe-O1-O2 of 43.93º and 51.90º around the iron-oxygen O1 bond, respectively. The water molecules bind to heme via 13 hydrogen bonds and O-H O and 6 C-H O interactions, which accounts for -67.23 kcal mol-1. It is observed that the inclusion of water molecules does not affect significantly the stability of the heme-artemisinin complex.