Seasonal Stability in the Microbiomes of Temperate Gorgonians and the Red Coral Corallium rubrum Across the Mediterranean Sea.

Populations of key benthic habitat-forming octocoral species have declined significantly in the Mediterranean Sea due to mass mortality events caused by microbial disease outbreaks linked to high summer seawater temperatures. Recently, we showed that the microbial communities of these octocorals are relatively structured; however, our knowledge on the seasonal dynamics of these microbiomes is still limited. To investigate their seasonal stability, we collected four soft gorgonian species (Eunicella singularis, Eunicella cavolini, Eunicella verrucosa and Leptogorgia sarmentosa) and the precious red coral (Corallium rubrum) from two coastal locations with different terrestrial impact levels in the Mediterranean Sea, and used next-generation amplicon sequencing of the 16S rRNA gene. The microbiomes of all soft gorgonian species were dominated by the same 'core microbiome' bacteria belonging to the Endozoicomonas and the Cellvibrionales clade BD1-7, whereas the red coral microbiome was primarily composed of 'core' Spirochaetes, Oceanospirillales ME2 and Parcubacteria. The associations with these bacterial taxa were relatively consistent over time at each location for each octocoral species. However, differences in microbiome composition and seasonal dynamics were observed between locations and could primarily be attributed to locally variant bacteria. Overall, our data provide further evidence of the intricate symbiotic relationships that exist between Mediterranean octocorals and their associated microbes, which are ancient and highly conserved over both space and time, and suggest regulation of the microbiome composition by the host, depending on local conditions.

Changes in genetic structure of Posidonia oceanica at Monterosso al Mare (Ligurian Sea) and its resilience over a decade (1998-2009).

Genetic differences in the Posidonia oceanica meadow of Monterosso al Mare (NW Mediterranean, Marine Protected Areas (MPAs) "Cinque Terre") were compared in three stations, at an increasing distance from a source of impact (beach nourishment) in the recent decade. Random amplified polymorphic DNA analysis showed a higher genetic variability (>20 %) in the area directly subjected to the stress, increasing with time. Clone integration, confirmed by phenotypic analysis, showed increases both in shoot density and leaf length connected to genetic differences observed in DNA fingerprints of new shoots. Analysis of molecular variance (AMOVA) showed 45 % individual differences within populations and 54 % among the populations. The fixation index (F (ST) = 0.54), of the genetic differentiation, showed a marked difference between the populations at different temporal scales. Over a decade AMOVA indicated genetic variations from 28 % (1998) to 54 % (2009). These results make it clear that in the P. oceanica population examined the environment had, in ten years, selected those clones which were more resistant to the anthropogenic impact, despite being subjected to the effects of the resuspension of fine sediments. These findings could help to explain both the survival of the regressed Mediterranean P. oceanica meadows in areas subjected to moderate impacts and the extreme variability in success of revegetation experiments. Management of the ecological disturbance here described indicates also the timescale in population response to stress and its increased resilience in MPAs.

Long-term High Resolution Image Dataset of Antarctic Coastal Benthic Fauna.

Antarctica is a remote place, the continent is covered by ice and its surrounding coastal areas are frozen for the majority of the year. Due to its peculiarity the observation of the underwater organisms is particularly difficult, complicated by logistic factors. We present a long-term dataset consisting of 755 images acquired by using a non-invasive, autonomous imaging device and encompassing both the Antarctic daylight and dark periods, including the corresponding transition phases. All images have the same field of view showing the benthic fauna and part of the water column above, including fishes present in the monitored period. All the images are manually annotated after a visual inspection performed by expert biologists. The extended monitoring period and the annotated images make the dataset a valuable benchmark suitable for studying the dynamics of the long-term Antarctic underwater fauna as well as for developing and testing algorithms for automated image analysis focused on the recognition and classification of the Antarctic organisms and the automated analysis of their long-term dynamics.

Effects of climate, invasive species and anthropogenic impacts on the growth of the seagrass Posidonia oceanica (L.) Delile in Liguria (NW Mediterranean Sea).

Eight shallow water Posidonia oceanica meadows were sampled in June 1999 along 300 km of the Ligurian coast and were compared through shoot density and lepidochronology. The growth of the seagrass was examined in the light of climate fluctuations and local stresses, colonisation by alien, invasive alga Caulerpa taxifolia, and effects of the oil spill from the tanker "Haven", and other anthropogenic impacts. Both shoot density and lepidochronology pointed to a generalised state of regression of all the meadows. The analysis of long-term growth curves of the rhizomes showed a positive trend parallel to the increase of air temperature. Two main groups of meadows were individuated on the basis of growth curve similarity. The first included four meadows, namely Ventimiglia, Imperia, Noli and Prelo, that were characterised by average values of rhizome growth of 8-9.1 mmyear(-1) and shoot density greater than 200 shootsm(-2). Although the Imperia meadow was the only one where the alien invasive alga C. taxifolia was found, it did not show differences for rhizome growth in comparison to the other meadows. The second group was formed by meadows that had suffered past anthropogenic impacts: Arenzano and Monterosso al Mare. They showed higher rhizome growth rates (9.4-10.6 mmyear(-1)) and shoot densities between 200 and 100 shootsm(-2). At Arenzano, where "Haven" oil was stranded in April 1991, no rhizome older than 8 years was found, thus confirming the shoot mortality induced by the oil spill event. The two last meadows exhibited growth curves very different from all the others: Portovenere, is a shallow meadow where P. oceanica merely survives in an extremely degraded situation with highest rhizome growth rate (12 mmyear(-1)), the other, Riva Trigoso, is the only meadow implanted on rock and had the lowest growth rates (7.1 mm year(-1)).

Genetic diversity and structure in two protected Posidonia oceanica meadows.

Posidonia oceanica meadows growing along the west Mediterranean coastline are under continuous anthropogenic pressure. The way meadow health correlates with genetic and genotypic diversity in P. oceanica, is still under debate. Here we report a microsatellite analysis of two P. oceanica meadows living in protected areas of the Ligurian (Monterosso al Mare, MPA of "Cinque Terre") and central Tyrrhenian Sea (Santa Marinella, regional Site of Community Importance). Both meadows were recently classified as "disturbed", according to shoot density and other phenological parameters. Between the two meadows, Santa Marinella showed higher genetic diversity, while clear genetic substructure was present in both sites, reflecting high spatial heterogeneity. The present study suggests that genetic diversity does not match unequivocally with shoot density and leaf morphology and that small scale intra-meadow heterogeneity is an important factor to consider for establishing the relation between genetic/genotypic variability and health of natural seagrass meadows.

Ecological change, sliding baselines and the importance of historical data: lessons from Combining [corrected] observational and quantitative data on a temperate reef over 70 years.

Understanding the effects of environmental change on ecosystems requires the identification of baselines that may act as reference conditions. However, the continuous change of these references challenges our ability to define the true natural status of ecosystems. The so-called sliding baseline syndrome can be overcome through the analysis of quantitative time series, which are, however, extremely rare. Here we show how combining historical quantitative data with descriptive 'naturalistic' information arranged in a chronological chain allows highlighting long-term trends and can be used to inform present conservation schemes. We analysed the long-term change of a coralligenous reef, a marine habitat endemic to the Mediterranean Sea. The coralligenous assemblages of Mesco Reef (Ligurian Sea, NW Mediterranean) have been studied, although discontinuously, since 1937 thus making available both detailed descriptive information and scanty quantitative data: while the former was useful to understand the natural history of the ecosystem, the analysis of the latter was of paramount importance to provide a formal measure of change over time. Epibenthic assemblages remained comparatively stable until the 1990s, when species replacement, invasion by alien algae, and biotic homogenisation occurred within few years, leading to a new and completely different ecosystem state. The shift experienced by the coralligenous assemblages of Mesco Reef was probably induced by a combination of seawater warming and local human pressures, the latter mainly resulting in increased water turbidity; in turn, cumulative stress may have favoured the establishment of alien species. This study showed that the combined analysis of quantitative and descriptive historical data represent a precious knowledge to understand ecosystem trends over time and provide help to identify baselines for ecological management.

A chromogenic substrate method for detecting and titrating anti-factor VIII antibodies in the presence of lupus anticoagulant.

BACKGROUND AND OBJECTIVES: The development of neutralizing anti-factor VIII antibodies (a-fVIII) is a major clinical complication. Lupus anticoagulant (LA) might affect detection of a-fVIII, since both inhibitors may act on the same coagulation pathway. Our aim was to accomplish unequivocal detection and titration of a-fVIII even in the presence of LA. DESIGN AND METHODS: We evaluated a-fVIII activity by a chromogenic substrate (CS) method in samples with a-fVIII (n=6), LA (n=12) and presumably both LA+a-fVIII (n=5). The inhibition index before (Ii) and after incubation at 37 C (Ii(37)) was estimated. We also performed factor VIII assays (one-stage and CS) and titration methods (Bethesda and CS) in parallel. RESULTS: Inhibition in the a-fVIII group (Ii=5-3200) was potentiated by incubation (Ii(37)=27-5200) as it was in LA+a-fVIII (Ii=9-21; Ii(37)=50-903). LA samples showed no or meaningless inhibitory effect (Ii=0-7; Ii(37)=0-4) or a-fVIII activity (0.00-0.06 CSU/ml) by the CS method; on the contrary, very low to moderate (0.52-7.00 BU/ml) a-fVIII activity was recorded by the Bethesda method. The two titration methods did not correlate (p>0.100) in the presence of LA, or LA+a-fVIII. Differences between factor VIII:C and factor VIIIcs were significant only in LA samples (p=0.005); however, patients with residual factor VIII activity from the LA+a-fVIII group also showed higher factor VIIIcs values than factor VIII:C ones. INTERPRETATION AND CONCLUSIONS: Results indicate the possibility of detecting and titrating a-fVIII without interference of LA by the CS method. This marks a difference with respect to the Bethesda method, in which a measurable effect can be expected in the presence of a strong LA.

Antiphospholipid antibodies impact the protein C (PC) pathway behavior.

Antiphospholipid antibodies may interfere with the PC pathway, displaying a resistance to the activated PC (resistant phenotype). This effect was evaluated by the APCR and the ProCG systems in 36 lupus anticoagulant samples, yielding abnormal results in 47% of APCR(original), 17% of APCR(modified), and 22% of ProCG test. ProCG values correlated with APCR(original) but not with APCR(modified). Most of lupus anticoagulants affecting the PC pathway showed abnormal APCR(original) results but not abnormal ProCG values. The different behavior between both systems may be due to the heterogeneity of the antibodies or could be attributed to the fact that, in the ProCG, a PC activator is added, while the APCR employs already activated exogenous PC.

Efecto del inhibidor lúpico o anti-factor VIII sobre la actividad del factor VIII-sustrato cromogénico / Lupus anticoagulant or anti-factor VIII effect on factor VIII activity-chromogenic substrate

El objetivo del trabajo fue analizar el efecto de inmunoglobulinas (IgG) purificadas con actividad anti-factor VIII neutralizante o inhibidor lúpico, sobre el factor VIII medido por sustrato cromogénico (factor VIII SC), con el propósito de validar el método para detectar anti-factor VIII en presencia de inhibidor lúpico. Se aisló la fracción IgG (cromatografía de afinidad) a partir de plasmas normales (IgG-N), con anti-factor VIII (IgG-aFVIII) o con inhibidor lúpico (IgG-IL), preparándose un pool normal como aporte de factor VIII en los ensayos de inhibición. Se analizaron las mezclas del pool normal con IgG-N, IgG-aFVIII, IgG-IL o la combinación de IgG-aFVIII+IgG-IL, inmediatamente y luego de 2 h a 37 ºC, determinándose la actividad del factor VIII SC y el índice de inhibición inmediato (Ii) o post-incubación (Ip). La inhibición y la potenciación producida por IgG-aFVIII+IgG-IL (Ii:30%±0,7; Ip:55%±2,7), fueron similares a las observadas con IgG-aFVIII sola (Ii:30%±0,6; Ip:55%±2,7). La IgG-IL no afectó la actividad de factor VIII SC. Semejante a lo observado en plasma, la IgG-IL no interfirió en la inhibición del factor VIII SC mediada por IgG-aFVIII. Se confirmó así la especificidad del método amidolítico, que permite detectar inhibidores anti-factor VIII independientemente de la coexistencia con el inhibidor lúpico, solucionando el problema diagnóstico planteado en hemofilia A.

To analyse the effect of purified immunoglobulins (IgG) with anti-factor VIII or lupus anticoagulant activity on factor VIII measured by chromogenic substrate (factor VIIICS), in order to validate the chromogenic substrate method for detection of anti-factor VIII activity in the presence of lupus anticoagulant. IgG fractions were purified (affinity chromatography) from normal plasmas (IgG-N) and plasmas with anti-factor VIII (IgG-aFVIII) or lupus anticoagulant (IgG-LA). A normal plasma pool was prepared as source of factor VIII in the inhibition tests. Mixtures of this pool with IgG-N, IgG-aFVIII, IgG-LA or IgG-aFVIII+IgG-LA were tested immediately or after 2 h at 37 °C by factor VIIICS method; inhibition index (Ii) and post-incubation index (Ip) were calculated. The inhibitory and progressive inhibitory effects produced by IgG-aFVIII+IgG-LA (Ii:30%±0.7; Ip:55%±2.7) were similar to those observed with IgG-aFVIII (Ii:30%±0.6; Ip:55%±2.7). The IgG-LA did not inhibit the factor VIIIcs activity. As was observed in plasma samples, IgG-aFVIII and the mixture of IgG-aFVIII+IgG-LA inhibited factor VIIICS, whereas IgG-LA did not. These results confirm the specificity of the amidolytic method in detecting anti-factor VIII inhibitors independently of the presence of lupus anticoagulant, thus solving the diagnostic problem in haemophilia A.

La nación en peligro: el debate médico sobre el aborto en Chile en la década de 1930 / The nation in danger: the medical debate about the abortion in Chile during the decade of 1930

Presenta el debate médico sobre el aborto en la década de 1930. Los médicos no fueron los únicos en Chile se referieron al 'aborto criminal', pero tuvieron la principal influencia mediática y política para mostrarlo ante la sociedad. El constacto profesional con las mujeres y, en gran medida, la progresiva participación y autoridad que asumían en los asuntos públicos contribuyeron a convertilos en cruciales forjadores de la noción de que el aborto era un flagelo nacional, objeto de imperativa atención estatal.

La Nación en peligro: el debate médico sobre el aborto en Chile en la década de 1930 / The Nation in danger: the medical debate about the abortion in Chile during the decade of 1930

Presenta el debate médico sobre el aborto en la década de 1930. Los médicos no fueron los únicos en Chile se referieron al 'aborto criminal', pero tuvieron la principal influencia mediática y política para mostrarlo ante la sociedad. El constacto profesional con las mujeres y, en gran medida, la progresiva participación y autoridad que asumían en los asuntos públicos contribuyeron a convertilos en cruciales forjadores de la noción de que el aborto era un flagelo nacional, objeto de imperativa atención estatal. [AU]