Age-associated changes in long-chain fatty acid profile during healthy aging promote pro-inflammatory monocyte polarization via PPARγ

Differences in lipid metabolism associate with age-related disease development and lifespan. Inflammation is a common link between metabolic dysregulation and aging. Saturated fatty acids (FAs) initiate pro-inflammatory signalling from many cells including monocytes; however, no existing studies have quantified age-associated changes in individual FAs in relation to inflammatory phenotype. Therefore, we have determined the plasma concentrations of distinct FAs by gas chromatography in 26 healthy younger individuals (age < 30 years) and 21 healthy FA individuals (age > 50 years). Linear mixed models were used to explore the association between circulating FAs, age and cytokines. We showed that plasma saturated, poly- and mono-unsaturated FAs increase with age. Circulating TNF-α and IL-6 concentrations increased with age, whereas IL-10 and TGF-β1 concentrations decreased. Oxidation of MitoSOX Red was higher in leucocytes from FA adults, and plasma oxidized glutathione concentrations were higher. There was significant colinearity between plasma saturated FAs, indicative of their metabolic relationships. Higher levels of the saturated FAs C18:0 and C24:0 were associated with lower TGF-β1 concentrations, and higher C16:0 were associated with higher TNF-α concentrations. We further examined effects of the aging FA profile on monocyte polarization and metabolism in THP1 monocytes. Monocytes preincubated with C16:0 increased secretion of pro-inflammatory cytokines in response to phorbol myristate acetate-induced differentiation through ceramide-dependent inhibition of PPARγ activity. Conversely, C18:1 primed a pro-resolving macrophage which was PPARγ dependent and ceramide dependent and which required oxidative phosphorylation. These data suggest that a midlife adult FA profile impairs the switch from proinflammatory to lower energy, requiring anti-inflammatory macrophages through metabolic reprogramming.

Age-associated changes in long-chain fatty acid profile during healthy aging promote pro-inflammatory monocyte polarization via PPARγ

Differences in lipid metabolism associate with age-related disease development and lifespan. Inflammation is a common link between metabolic dysregulation and aging. Saturated fatty acids (FAs) initiate pro-inflammatory signalling from many cells including monocytes; however, no existing studies have quantified age-associated changes in individual FAs in relation to inflammatory phenotype. Therefore, we have determined the plasma concentrations of distinct FAs by gas chromatography in 26 healthy younger individuals (age < 30 years) and 21 healthy FA individuals (age > 50 years). Linear mixed models were used to explore the association between circulating FAs, age and cytokines. We showed that plasma saturated, poly- and mono-unsaturated FAs increase with age. Circulating TNF-α and IL-6 concentrations increased with age, whereas IL-10 and TGF-β1 concentrations decreased. Oxidation of MitoSOX Red was higher in leucocytes from FA adults, and plasma oxidized glutathione concentrations were higher. There was significant colinearity between plasma saturated FAs, indicative of their metabolic relationships. Higher levels of the saturated FAs C18:0 and C24:0 were associated with lower TGF-β1 concentrations, and higher C16:0 were associated with higher TNF-α concentrations. We further examined effects of the aging FA profile on monocyte polarization and metabolism in THP1 monocytes. Monocytes preincubated with C16:0 increased secretion of pro-inflammatory cytokines in response to phorbol myristate acetate-induced differentiation through ceramide-dependent inhibition of PPARγ activity. Conversely, C18:1 primed a pro-resolving macrophage which was PPARγ dependent and ceramide dependent and which required oxidative phosphorylation. These data suggest that a midlife adult FA profile impairs the switch from proinflammatory to lower energy, requiring anti-inflammatory macrophages through metabolic reprogramming.

Characterization of microvesicles released from human red blood cells

Background/Aims: Extracellular vesicles (EVs) are spherical fragments of cell membrane released from various cell types under physiological as well as pathological conditions. Based on their size and origin, EVs are classified as exosome, microvesicles (MVs) and apoptotic bodies. Recently, the release of MVs from human red blood cells (RBCs) under different conditions has been reported. MVs are released by outward budding and fission of the plasma membrane. However, the outward budding process itself, the release of MVs and the physical properties of these MVs have not been well investigated. The aim of this study is to investigate the formation process, isolation and characterization of MVs released from RBCs under conditions of stimulating Ca2+ uptake and activation of protein kinase C. Methods: Experiments were performed based on single cell fluorescence imaging, fluorescence activated cell sorter/flow cytometer (FACS), scanning electron microscopy (SEM), atomic force microscopy (AFM) and dynamic light scattering (DLS). The released MVs were collected by differential centrifugation and characterized in both their size and zeta potential. Results: Treatment of RBCs with 4-bromo-A23187 (positive control), lysophosphatidic acid (LPA), or phorbol-12 myristate-13 acetate (PMA) in the presence of 2 mM extracellular Ca2+ led to an alteration of cell volume and cell morphology. In stimulated RBCs, exposure of phosphatidylserine (PS) and formation of MVs were observed by using annexin V-FITC. The shedding of MVs was also observed in the case of PMA treatment in the absence of Ca2+, especially under the transmitted bright field illumination. By using SEM, AFM and DLS the morphology and size of stimulated RBCs, MVs were characterized. The sizes of the two populations of MVs were 205.8 ± 51.4 nm and 125.6 ± 31.4 nm, respectively. Adhesion of stimulated RBCs and MVs was observed. The zeta potential of MVs was determined in the range from - 40 mV to - 10 mV depended on the solutions and buffers used. Conclusion: An increase of intracellular Ca2+ or an activation of protein kinase C leads to the formation and release of MVs in human RBCs.

(Table 5, page 626), Tin and main metal elements composition of ferromanganese concretions (acid-soluble fractions)

Concentrations of tin in sea water decreased from estuarine and shelf (0.02-0.04 µg/kg) to surface Atlantic waters (0.009 µg/kg). Mean contents (ppm) in other materials included: ultramafic rocks, 0.8; basalts, 1.7; silicic rocks, 2.5; red clays, 3.4; amphibolites, 1.2. Oceanic ferromanganese deposits contained from 0.2 to 5.8 ppm; tin and cobalt contents were correlated.

Data compilation on the biological response to ocean acidification: environmental and experimental context of data sets and related literature

The exponential growth of studies on the biological response to ocean acidification over the last few decades has generated a large amount of data. To facilitate data comparison, a data compilation hosted at the data publisher PANGAEA was initiated in 2008 and is updated on a regular basis (doi:10.1594/PANGAEA.149999). By January 2015, a total of 581 data sets (over 4 000 000 data points) from 539 papers had been archived. Here we present the developments of this data compilation five years since its first description by Nisumaa et al. (2010). Most of study sites from which data archived are still in the Northern Hemisphere and the number of archived data from studies from the Southern Hemisphere and polar oceans are still relatively low. Data from 60 studies that investigated the response of a mix of organisms or natural communities were all added after 2010, indicating a welcomed shift from the study of individual organisms to communities and ecosystems. The initial imbalance of considerably more data archived on calcification and primary production than on other processes has improved. There is also a clear tendency towards more data archived from multifactorial studies after 2010. For easier and more effective access to ocean acidification data, the ocean acidification community is strongly encouraged to contribute to the data archiving effort, and help develop standard vocabularies describing the variables and define best practices for archiving ocean acidification data.

(Table 2) Fatty acid composition of polychaetes sampled during POLARSTERN cruise ANT-XXIV/2

The fatty acid (FA) composition of representatives belonging to 18 polychaete families from the Southern Ocean shelf and deep sea (600 to 5337 m) was analysed in order to identify trophic biomarkers and elucidate possible feeding preferences. Total FA content was relatively low with few exceptions and ranged from 1.0 to 11.6% of total body dry weight. The most prominent FA found were 20:5(n-3), 16:0, 22:6(n-3), 18:1(n-7), 20:4(n-6), 18:0, 20:1(n-11) and 18:1(n-9). For some polychaete families and species FA profiles indicated selective feeding on certain dietary components, like freshly deposited diatom remains (e.g., Spionidae, Fauveliopsidae and Flabelligeridae) or foraminiferans (e.g., Euphrosinidae, Nephtyidae and Syllidae). Feeding patterns were relatively consistent within families at the deep stations, while the FA composition differed between the deep and the shelf stations within the same family. Fatty alcohols, indicative of wax ester storage, were found in almost all families (in proportions of 0.0 to 29.3% of total FA and fatty alcohols). The development of this long-term storage mechanism of energy reserves possibly displays an evolutionary strategy.

Stable isotope ratios of a oyster Hyotissa hyotis from the northern Red Sea

This study explores the giant oyster Hyotissa hyotis as a novel environmental archive in tropical reef environments of the Indo-Pacific. The species is a typical accessory component in coral reefs, can reach sizes of tens of centimetres, and dates back to the Late Pleistocene. Here, a 70.2-mm-long oxygen and carbon isotope transect through the shell of a specimen collected at Safaga Bay, northern Red Sea, in May 1996, is presented. The transect runs perpendicularly to the foliate and vesicular layers of the inner ostracum near the ligament area of the oyster. The measured d18O and d13C records show sinusoidal fluctuations, which are independent of shell microstructure. The d13C fluctuations exhibit the same wavelength as the d18O fluctuations but are phase shifted. The d18O record reflects the sea surface temperature variations from 1957 until 1996, possibly additionally influenced by the local evaporation. Due to locally enhanced evaporation in the semi-enclosed Safaga Bay, the d18Oseawater value is estimated at 2.17 per mil, i.e., 0.3-0.8 per mil higher than published open surface water d18O values (1.36-1.85 per mil) from the region. The mean water temperature deviates by only 0.4°C from the expected value, and the minimum and maximum values are 0.5°C lower and 2.9°C higher, respectively. When comparing the mean monthly values, however, the sea surface temperature discrepancy between reconstructed and global grid datasets is always <1.0°C. The d13C signal is weakly negatively correlated with regional chlorophyll a concentration and with the sunshine duration, which may reflect changes in the bivalve's respiration. The study emphasises the palaeogeographic context in isotope studies based on fossils, because coastal embayments might not reflect open-water oceanographic conditions.

Elemental microchemistry, fatty acid profile and geometric morphometrics signatures of goose barnacles (Pollicipes pollicipes) reveal their place of origin

Os produtos provenientes do mar têm um importante papel a nível socioeconómico, gastronómico e no legado cultural das comunidades piscatórias e costeiras de Portugal. O Percebe Pollicipes pollicipes é na Península Ibérica o recurso biológico do intertidal mais explorado pelo ser humano, devido à sua enorme popularidade e consequente procura, fazendo em algumas ocasiões disparar o preço de mercado até 150€/kg, gerando na sobreexploração dos stocks existentes. No entanto, na área marinha protegida da Reserva Natural da Berlenga, a apanha do percebe é fortemente regulada, tendo-se tornado em Portugal num bom exemplo da gestão de recursos marinhos. Com o intuito de prevenir fraudes, adulteração alimentar ou quaisquer outras práticas que possam induzir o consumidor em erro a Comissão Europeia declara que, o consumidor tem o direito de receber informação correcta acerca dos produtos que adquire, para além de definir regras para a correcta aplicação destas regras. Métodos analíticos que possibilitem identificar a origem do percebe, tornam-se deste modo importantes ferramantas no desenvolvimento de um selo de Denominação de Origem Protegida (DOP) e na gestão comercial do produto. Deste modo, investigou-se se o Percebe possui diferenças específicas de cada local de captura, através da forma da unha (CS), da composição microquímica da unha (EM) e do perfil de ácidos gordos (FA). A análise foi efectuada em indivíduos recolhidos em 3 locais na Reserva Natural das Berlengas e 7 ao longo de 300 km da costa Portuguesa. Em cada indivíduo analisou-se a forma da unha (CS) através da morfometria geométrica, a composição microquímica da unha (EM) através de ICP-MS e o perfil de ácidos gordos do músculo através de GC-FID. A análise das funções discriminantes (DFA) quer para a EM quer para a FA em separado obteve um elevado sucesso de reclassificação (77,6 % e 99% respectivamente, através de validação cruzada), enquanto que EM combinado com FA permitiu um sucesso de reclassificação de 100 %. A análise discriminante baseada apenas na CS, demonstrou um baixo sucesso (29,6 %) .Estes resultados demonstram que a composição microquímica da unha e o perfil de ácidos gordos do músculo de Percebe, poderá ser uma ferramenta de elevada importância, na determinação da origem do Percebe. Esta abordagem poderá ser utilizada para identificar a origem dos percebes comercializados, bem como ajudar no desenvolvimento de um selo DOP, aumentando ao mesmo tempo o valor potencial dos recursos biológicos provenientes de áreas marinhas protegidas em Portugal.

Age determination and clay layer frequencies of sediments of the Black and Red Sea

Paleoenvironmental proxy data for ocean properties, eolian sediment input, and continental rainfall based on high-resolution analyses of sediment cores from the southwestern Black Sea and the northernmost Gulf of Aqaba were used to infer hydroclimatic changes in northern Anatolia and the northern Red Sea region during the last ~7500 years. Pronounced and coherent multicentennial variations in these records reveal patterns that strongly resemble modern temperature and rainfall anomalies related to the Arctic Oscillation/North Atlantic Oscillation (AO/NAO). These patterns suggest a prominent role of AO/NAO-like atmospheric variability during the Holocene beyond interannual to interdecadal timescales, most likely originating from solar output changes.

Benthic foraminifera of the Red Sea

The distribution of living (Rose Bengal-stained), dead and fossil benthic foraminifera was investigated in six short cores (multicores, 30-32 cm total length) recovered from the central Red Sea. The ecological preferences as well as the relationship between the live and dead/fossil assemblages (preserved down-core) were examined. The sites, located along a W-E profile and between the depth of 366 and 1782 m, extend from the center of the oxygen minimum zone (OMZ, ~200-650 m), through its margin at ~600 m, and down to the well-aerated deep-water environment. Live (Rose-Bengal stained) and coexisting dead foraminifera were studied in the upper 5 cm of each of the sites, and the fossil record was studied down to ~32 cm. Q-mode Principal Component Analysis was used and four distinct foraminiferal fossil assemblages were determined. These assemblages follow different water mass properties. In the center of the OMZ, where the organic carbon content is highest and the oxygen concentration is lowest (<=0.5 ml O2/l), the Bolivina persiensis-Bulimina marginata-Discorbinella rhodiensis assemblage dominates. The slightly more aerated and lower organic-carbon-content seafloor, at the margin of the OMZ, is characterized by the Neouvigerina porrecta-Gyroidinoides cf. G. soldanii assemblage. The transitional environment, between 900-1200 m, with its well-aerated and oligotrophic seafloor, is dominated by the Neouvigerina ampullacea-Cibicides mabahethi assemblage. The deeper water (>1500 m), characterized by the most oxygenated and oligotrophic seafloor conditions, is associated with the Astrononion sp. A-Hanzawaia sp. A assemblage. Throughout the Red Sea extremely high values of temperature and salinity are constant below ~200 m depth, but the flux of organic matter to the sea floor varies considerably with bathymetry and appears to be the main controlling factor governing the distribution pattern of the benthic foraminifera. Comparison between live and the dead/fossil assemblages reveals a large difference between the two. Processes that may control this difference include species-specific high turnover rates, and preferential predation and loss of fragile taxa (either by chemical or microbial processes). Significant variations in the degree of loss of the organic-cemented agglutinants were observed down core. This group is preserved down to 5-10 cm at the shallow OMZ sites and down to greater depths at well-aerated and oligotrophic sites. The lower rate of disintegration of these forms, in the deeper locations of the Red Sea, may be related to low microbial activity. This results in the preservation of increasing numbers of organic-cemented shells down-core.

(Table 2) Age determination of brock-red sandy mud beds and detrital carbonate beds in the Scotian margin

Piston cores from the continental margin off Nova Scotia show up to four discrete intervals of "brick-red sandy mud", which are up to 20 cm thick. The ages of these intervals are bracketed by several radiocarbon dates, and three fall in the range 12.5-14.1 ka (radiocarbon years with -0.4 kyr reservoir correction). The youngest dates from ~10.4 ka, placing it within the Younger Dryas. The distribution of the beds and their petrographic character indicate a source in the Gulf of Saint Lawrence. The grain size of these beds suggests that they comprise a coarse component transported by ice rafting that diminishes distally and a fine component that represents suspension fallout from a surface plume and resulting nepheloid layers. Graded brick-red beds in some cores were probably redeposited from turbidity currents. The lowermost bed on the Laurentian Fan and East Scotian Rise is immediately overlain by a carbonate-rich interval that can be identified all around the margin of the Grand Banks. This interval is correlated with detrital carbonate bed DC-1 in the Labrador Sea and Heinrich event H1 in the North Atlantic. The sequential occurrence of the two beds suggests that they may be a response to the same trigger, probably sea level rise, but that the Gulf of Saint Lawrence source was more easily destabilized.

Amino acid racemization for several DSDP samples (Table 1)

Kinetic parameters for the epimerization of isoleucine in multispecific foraminiferal asemblages were used to establish the effects of burial depth and the geothermal gradient on the extent of reaction. It was observed that with a little as thirty meters of burial in a normal thermal regime there were differences between the extent of epimerization measured and that which would have been predicted for thermal equilibrium with bottom water temperatures. As would be expected, these differences are greatest when the heat flow (the geothermal gradient) and/or the sedimentation rates are highest. These effects were observed in most of the DSDP samples studied, and have been used to estimate the average heat flux since the time of sample deposition. Occasional anomalous effects were observed which could not be related to past or present heat flux. These were determined to be due to such geologic occurrences as slumping and reworking or to recent sample contamination. Other problems emerged related to bottom water temperatures including changes over geologic time which are unknown and could not be deduced. Thus, the presence of epimerization anomalies in DSDP cores as noted above limits the effectiveness of amino acid geochronology in such cores, unless these anomalies can be recognized as ab initio.

Seawater carbonate chemistry, calcification and dissolution of a coral reef in the northern Red Sea, 2007

In this study we investigated the relations between community calcification of an entire coral reef in the northern Red Sea and annual changes in temperature, aragonite saturation and nutrient loading over a two year period. Summer (April-October) and winter (November-March) average calcification rates varied between 60 ± 20 and 30 ± 20 mmol·m-2·d-1, respectively. In general, calcification increased with temperature and aragonite saturation state of reef water with an apparent effect of nutrients, which is in agreement with most laboratory studies and in situ measurements of single coral growth rates. The calcification rates we measured in the reef correlated remarkably well with precipitation rates of inorganic aragonite calculated for the same temperature and degree of saturation ranges using empirical equations from the literature. This is a very significant finding considering that only a minute portion of reef calcification is inorganic. Hence, these relations could be used to predict the response of coral reefs to ocean acidification and warming.

EXAMINATION OF THE EFFECTS OF VALPROIC ACID EXPOSURE OR P300 PROTEIN INHIBITION ON THE BALANCE OF APOPTOSIS AND PROLIFERATION IN P19 EMBRYONAL CARCINOMA CELLS

Valproic acid (VPA), a commonly-used anticonvulsant drug, is associated with increased risk of fetal malformations, including neural tube defects (NTDs). Previous in vivo studies determined that VPA-exposed embryos with a NTD had altered expression of several proteins regulated by p300, a histone acetyltransferase (HAT) protein. p300 is capable of acetylating histones and non-histone proteins through its HAT activity, allowing it to transcriptionally regulate genes as well as modulate the stability and activity of specific proteins. NFκB, Stat3 and Egr1, all of which function as transcription factors, are regulated by p300 through its HAT activity. Together, these proteins all play an important role in maintaining the balance of apoptosis, proliferation and differentiation, the regulation of which is extremely important for proper embryonic development. The studies in this thesis utilized P19 embryonal carcinoma (EC) cells in order to determine the effects of VPA exposure on the expression of p300 and the aforementioned transcription factors, as well as apoptosis and proliferation, in vitro. P19 EC cells were exposed to C646, a selective p300 inhibitor, in order to assess whether the effects observed as a result of VPA exposure were due to p300 protein degradation. It was found that VPA exposure for 24 hours in P19 EC cells in vitro resulted in a significant decrease in p300 protein expression. VPA exposure also significantly decreased NFκB protein expression, while resulting in increased Stat3 protein expression. However, Stat3 acetylation and phosphorylation, which both contribute to Stat3 activation, were significantly decreased as a result of VPA exposure. p300 inhibition resulted in a significant decrease in NFκB, similar to what was observed as a result of VPA exposure, which suggests that VPA-mediated degradation of p300 may play a role in reduced NFκB protein expression following VPA exposure. Conversely, Stat3 protein expression, acetylation and phosphorylation were not significantly changed as a result of p300 inhibition, suggesting that p300 degradation does not play a role in VPA’s effects on Stat3 protein expression and activation. VPA exposure also resulted in a significant increase in apoptosis, while p300 inhibition did not significantly increase apoptosis. These data suggest that p300 degradation plays a role in VPA-mediated teratogenicity, and that VPA may target other cellular mechanisms in order to exert its teratogenic effects.