Effects of ocean acidification and warming on the development and biochemical responses of juvenile shrimp Palaemon elegans (Rathke, 1837).

Anthropogenic CO2 emissions have led to the warming and acidification of the oceans. Although, there is a growing of evidence showing that simultaneous occurrence of ocean acidification and ocean warming are threats to marine organisms, information on their combined effect on coastal shrimp species remains scarce. The purpose of this study was to estimate the combined effects of seawater acidification and warming on growth-related traits and biochemical responses of P. elegans juveniles. In this work, shrimp were exposed for 65 days at 4 experimental conditions: pH 8.10 * 18 °C, pH 7.80 * 18 °C, pH 8.10 * 22 °C, pH 7.80 * 22 °C. The results showed that low pH decreases the lipid content by ∼13% (p < 0.05). Higher temperature reduced the condition factor by ∼11%, the protein content by ∼20%, the PUFA by ∼8,6% and shortened moulting events by 5 days (p > 0.05) while the SFA increased ∼9.4%. The decrease in condition factor and protein was however more prominent in organisms exposed to the combination of pH and temperature with a decrease of ∼13% and ∼21%, respectively. Furthermore, essential fatty acids as EPA and DHA also decreased by ∼20% and ∼6.6% in low pH and higher temperature condition. Despite this study suggest that warming may have a greater impact than acidification, it has been shown that their combined effect can exacerbate these impacts with consequences for the shrimp's body size and biochemical profile.

Ecological features of a rocky intertidal community exposed to sewage effluent.

Sewage is among the largest components of coastal pollution, showing a variable scale and size when causing an impact. In this study, temporal and spatial sewage-related gradients were identified using univariate and multivariate methods. Phosphates and nitrogen-based nutrients, except nitrate, were associated to sewage. Abundance and species richness decrease with distance from the sewage outfall while evenness increased. Filter-feeder bivalves and grazer crustaceans showed preference for the sewage discharge site. Gastropods were more abundant at an intermediate distance, likely reflecting primary production enhancement by sewage. Beta diversity was lower at the location and time with highest sewage loadings. The turnover component of beta diversity expressed as an absolute or proportional value was also useful to detect temporal and spatial sewage-related gradients. Highly energetic hydrodynamics contributes to ameliorate sewage impacts, yet not enough to veil the effect of pollution in this study.

Comparative Methodological Assessment of the Randomized GLOBAL LEADERS Trial Using Total Ischemic and Bleeding Events.

BACKGROUND: Time-to-first-event analysis considers only the first event irrespective of its severity. There are several methods to assess trial outcomes beyond time-to-first-event analysis, such as analyzing total events and ranking outcomes. In the GLOBAL LEADERS study, time-to-first-event analysis did not show superiority of ticagrelor monotherapy following one-month dual antiplatelet therapy (DAPT) after percutaneous coronary intervention to conventional 12-month DAPT followed by aspirin monotherapy in the reduction of the primary composite end point of all-cause mortality or new Q-wave myocardial infarction. This study sought to explore various analytical approaches in assessing total ischemic and bleeding events after percutaneous coronary intervention in the GLOBAL LEADERS study. METHODS AND RESULTS: Total ischemic and bleeding events were defined as all-cause mortality, any stroke, any myocardial infarction, any revascularization, or Bleeding Academic Research Consortium grade 2 or 3 bleeding. We used various analytical approaches to analyze the benefit of ticagrelor monotherapy over conventional DAPT. For ischemic and bleeding events at 2 years after percutaneous coronary intervention, ticagrelor monotherapy demonstrated a 6% risk reduction, compared with conventional 12-month DAPT in time-to-first-event analysis (hazard ratio, 0.94 [95% CI, 0.88-1.01]; log-rank P=0.10). In win ratio analysis, win ratio was 1.05 (95% CI, 0.97-1.13; P=0.20). Negative binomial regression and Andersen-Gill analyses which include repeated events showed statistically significant advantage for ticagrelor monotherapy (rate ratio, 0.92 [95% CI, 0.85-0.99; P=0.020] and hazard ratio, 0.92 [95% CI, 0.85-0.99; P=0.028], respectively), although in weighted composite end point analysis, the hazard ratio was 0.93 (95% CI, 0.84-1.04; log-rank P=0.22). CONCLUSIONS: Statistical analyses considering repeated events or event severity showed that ticagrelor monotherapy consistently reduced ischemic and bleeding events by 5% to 8%, compared with conventional 1-year DAPT. Applying multiple statistical methods could emphasize the multiple facets of a trial and result in accurate and more appropriate analyses. Considering the recurrence of ischemic and bleeding events, ticagrelor monotherapy appeared to be beneficial after percutaneous coronary intervention. Registration: URL: https://www.clinicaltrials.gov; Unique identifier: NCT01813435.

Antifungal and Antioxidant Properties of Chitosan Polymers Obtained from Nontraditional Polybius henslowii Sources.

Chitin was extracted from Polybius henslowii, a swimming crab, captured in large quantities throughout the Portuguese coast by purse seine vessels as bycatch. After standard chitin extraction procedures, water-soluble chitosan products were obtained via two different methods: (1) N-acetylation with the addition of acetic anhydride and (2) a reaction with hydrogen peroxide. The chemical structure and molecular weight of chitosan derivatives, water-soluble chitosan (WSC) and chitooligosaccharides (COS), were confirmed by Fourier Transform Infrared Spectroscopy (FT-IR) and gel permeation chromatography (GPC). Antioxidant and metal chelation activities were evaluated, and the growth inhibition capacity was tested on four phytopatogens. The chitooligosaccharides from pereopods (pCOS) and shell body parts (sCOS) inhibited all fungal species tested, particularly Cryphonectria parasitica with 84.7% and 85.5%, respectively. Both radical scavenging and antifungal activities proved to be dose-dependent. Chitooligosaccharides with a low molecular weight (2.7, 7.4, and 10.4 Kg·mol-1) showed the highest activity among all properties tested. These results suggested that chitosan derivatives from P. henslowii raw material could potentially be used against phytopathogens or as ingredient in cosmetics and other products related to oxidative stress.

Comparação entre avaliação de isquemia miocárdica pela reserva de fluxo fracionada e cintilografia de perfusão miocárdica / Comparison between myocardial ischemia evaluation by fractional flow reserve and myocardial perfusion scintigraphy

Lesões coronárias moderadas podem ser, ou não, responsáveis pela isquemia miocárdica. A análise funcional das lesões pode ser realizada por métodos invasivos e não invasivos. Comparar a análise funcional das lesões coronarianas moderadas pela reserva de fluxo fracionado e pela cintilografia de perfusão miocárdica. Foram estudados prospectivamente 47 pacientes com doença arterial coronária estável com pelo menos uma lesão coronariana moderada obstrutiva. Eles foram submetidos à reserva de fluxo fracionado e à cintilografia de perfusão miocárdica com intervalo médio de 24,5 dias, entre janeiro de 2013 e dezembro de 2015. Não houve alteração no estado clínico e nem no procedimento de revascularização entre exames. As variáveis populacionais foram descritas como mediana e interquartil. A reserva de fluxo fracionado foi realizada em um de tronco de coronária esquerda; 37 artérias coronárias descendentes; 12 artérias circunflexas e quatro artérias coronárias direitas. Reserva de fluxo fracionado < 0,8 foi considerada positiva. A análise comparativa entre os resultados dos testes foi feita pelo teste de Fisher bicaudal, sendo considerado significativo valor de p < 0,05. A reserva de fluxo fracionado < 0,8 foi encontrada no tronco de coronária esquerda (100%); 13 na artéria coronária descendente (35,14%); seis na artéria circunflexa (50%) e duas na artéria coronária direita (50%). Dentre os pacientes com reserva de fluxo fracionado positiva, 83% tinham isquemia miocárdica demonstrada na cintilografia de perfusão miocárdica (p = 0,058). Analisando especificamente o território da artéria coronária descendente, 83% dos pacientes com reserva de fluxo fracionado negativa não tinham isquemia na cintilografia de perfusão miocárdica, mas 69% dos pacientes com reserva de fluxo fracionado positiva não tinham isquemia na cintilografia de perfusão miocárdica (p = 0,413). Pode ocorrer discordância entre os resultados de análise funcional de lesões coronárias moderadas por testes invasivos e não invasivos

Moderate coronary artery lesions can be, or not, responsible for myocardial ischemia. The functional analysis of these lesions can be performed by invasive and noninvasive methods.To compare the functional analysis of moderate coronary lesions by fractional flow reserve and myocardial perfusion scintigraphy. 47 patients with stable coronary artery disease and at least one moderate coronary artery obstruction were prospectively studied. They were submitted to fractional flow reserve and myocardial perfusion scintigraphy with a median interval of 24.5 days between January 2013 and December 2015. There was no change in clinical status or revascularization procedure between the exams. The population variables were described as medians and interquartile range. Fractional flow reserve was performed in one left main coronary artery; 37 left descending coronary arteries; 12 circumflex arteries and 4 right coronary arteries. Fractional flow reserve < 0.8 was considered positive. The comparative analysis between the results of the tests was performed by two-tailed Fisher's test and a p-value 0.05 was considered significant.Fractional flow reserve < 0.8 was found in the left main coronary artery (100%); 13 in the left descending coronary artery (35.14%); 6 in circumflex artery (50%) and 2 in the right coronary artery (50%). Among the patients with positive fractional flow reserve, 83% had myocardial ischemia demonstrated by the myocardial perfusion scintigraphy (p = 0.058).When analyzing specifically the left descending coronary artery, 83% of the patients with negative fractional flow reserve showed no ischemia at the myocardial perfusion scintigraphy, but 69% of the patients with positive fractional flow reserve showed no ischemia at the myocardial perfusion scintigraphy (p = 0.413). Disagreements can occur between the results of the functional analysis of moderate coronary lesions by invasive and noninvasive tests

Trophic links and nutritional condition of fish early life stages in a temperate estuary.

The physiological and nutritional condition of fish larvae affect their survival and thus, the success of estuaries as nursery areas. Fatty acid composition has been useful to determine fish nutritional condition, as well as trophic relationships in marine organisms. The present study analyses the fatty acid (FA) composition of fish larvae during spring and summer in the Mondego estuary, Portugal. FA composition, trophic markers (FATM) and fish nutritional condition was analysed for Gobiidae and Sardina pilchardus larvae and the relationships with the local environment evaluated. Results showed that both taxa differed mainly in the stearic acid (C18:0) and eicosapentaenoic acid (EPA) content, with important amounts in Gobiidae and S. pilchardus, respectively. Gobiidae larvae presenting high nutritional condition and omnivore FATM. Fatty acid composition seems to be related with their natural habitat selection and food availability, while fish larvae nutritional condition also showed a strong link with the water temperature and presence of potential predators. This study suggests that FA composition can be a useful tool in assessing planktonic trophic relationships and in identifying species natural habitat.

Homarus gammarus (Crustacea: Decapoda) larvae under an ocean acidification scenario: responses across different levels of biological organization.

The present study evaluated the effects of exposure to different target pCO2 levels: control (C: 370µatm, pH=8.15) and ocean acidification (OA: 710µatm, pH=7.85) on development and biochemical responses related with oxidative stress and energy metabolism during the crustacean Homarus gammarus (L.) larval development, integrating different levels of biological organization. After hatching in the laboratory, larvae from the same female brood were exposed to the described conditions from hatching until reaching Stage III (last larval stage - 11days). H. gammarus larvae demonstrated some susceptibility when addressing the predicted pCO2 levels for 2100. Further analysis at the biochemical and physiological level highlighted the occurrence of oxidative stress in the OA scenario (Superoxide Dismutase reduction and higher DNA damage) that was followed by developmental effects, increased inter-moult period from SII to SIII and reduced growth. The extended exposure to these conditions may affect organisms' key life-cycle functions such as physiological resistance, growth, sexual maturation, or reproduction with implications in their future fitness and population dynamics.

Analysis of Pelagia noctiluca proteome Reveals a Red Fluorescent Protein, a Zinc Metalloproteinase and a Peroxiredoxin.

Pelagia noctiluca is the most venomous jellyfish in the Mediterranean Sea where it forms dense blooms. Although there is several published research on this species, until now none of the works has been focused on a complete protein profile of the all body constituents of this organism. Here, we have performed a detailed proteomics characterization of the major protein components expressed by P. noctiluca. With that aim, we have considered the study of jellyfish proteins involved in defense, body constituents and metabolism, and furthered explore the significance and potential application of such bioactive molecules. P. noctiluca body proteins were separated by1D SDS-PAGE and 2DE followed by characterization by nanoLC-MS/MS and MALDI-TOF/TOF techniques. Altogether, both methods revealed 68 different proteins, including a Zinc Metalloproteinase, a Red Fluorescent Protein (RFP) and a Peroxiredoxin. These three proteins were identified for the first time in P. noctiluca. Zinc Metalloproteinase was previously reported in the venom of other jellyfish species. Besides the proteins described above, the other 65 proteins found in P. noctiluca body content were identified and associated with its clinical significance. Among all the proteins identified in this work we highlight: Zinc metalloproteinase, which has a ShK toxin domain and therefore should be implicated in the sting toxicity of P. noctiluca.; the RFP which are a very important family of proteins due to its possible application as molecular markers; and last but not least the discovery of a Peroxiredoxin in this organism makes it a new natural resource of antioxidant and anti-UV radiation agents.

Harvest locations of goose barnacles can be successfully discriminated using trace elemental signatures.

European Union regulations state that consumers must be rightfully informed about the provenance of fishery products to prevent fraudulent practices. However, mislabeling of the geographical origin is a common practice. It is therefore paramount to develop forensic methods that allow all players involved in the supply chain to accurately trace the origin of seafood. In this study, trace elemental signatures (TES) of the goose barnacle Pollicipes pollicipes, collected from ten sites along the Portuguese coast, were employed to discriminate individual's origin. Barium (Ba), boron (B), cadmium (Cd), chromium (Cr), lithium (Li), magnesium (Mg), manganese (Mn), phosphorous (P), lead (Pb), strontium (Sr) and zinc (Zn) - were quantified using Inductively Coupled Plasma-Mass Spectrometry (ICP-MS). Significant differences were recorded among locations for all elements. A regularized discriminant analysis (RDA) revealed that 83% of all individuals were correctly assigned. This study shows TES can be a reliable tool to confirm the geographic origin of goose barnacles at fine spatial resolution. Although additional studies are required to ascertain the reliability of TES on cooked specimens and the temporal stability of the signature, the approach holds great promise for the management of goose barnacles fisheries, enforcement of conservation policies and assurance in accurate labeling.

Biochemical performance of native and introduced clam species living in sympatry: The role of elements accumulation and partitioning.

The present study reports metal and arsenic contamination in sediments, as well as element accumulation and partitioning in native (Ruditapes decussatus and Venerupis corrugata) and introduced (Ruditapes philippinarum) clam species living in sympatry at the Óbidos lagoon (Portugal). The biochemical performance and the human health risks derived from the consumption of these species are also discussed. The results obtained showed that R. decussatus was the most abundant species in all the sampling sites, revealing that the introduced clam has not yet supplanted the native species. The concentration of elements was higher in areas with higher Total Organic Matter (TOM) and fines content, being Chromium (Cr), Copper (Cu) and Lead (Pb) the most abundant metals. Clams from these areas showed the highest concentration of elements but the lowest bioaccumulation levels. Furthermore, except for As, higher concentration of elements was found in clams insoluble fraction, the less toxic fraction to the organisms. Due to the low contamination levels and because elements, except As, were mainly allocated to the insoluble fraction, clams presented similar biochemical parameters among distinct areas, with no significant oxidative stress induced. Furthermore, clams from the Óbidos lagoon represent a low health risk to human consumption since, except for As, their contamination levels were below the maximum permissible limits defined by international organizations.

Gulls identified as major source of fecal pollution in coastal waters: a microbial source tracking study.

Gulls were reported as sources of fecal pollution in coastal environments and potential vectors of human infections. Microbial source tracking (MST) methods were rarely tested to identify this pollution origin. This study was conducted to ascertain the source of water fecal contamination in the Berlenga Island, Portugal. A total of 169 Escherichia coli isolates from human sewage, 423 isolates from gull feces and 334 water isolates were analyzed by BOX-PCR. An average correct classification of 79.3% was achieved. When an 85% similarity cutoff was applied 24% of water isolates were present in gull feces against 2.7% detected in sewage. Jackknifing resulted in 29.3% of water isolates classified as gull, and 10.8% classified as human. Results indicate that gulls constitute a major source of water contamination in the Berlenga Island. This study validated a methodology to differentiate human and gull fecal pollution sources in a real case of a contaminated beach.

Estudo histomorfométrico da interface óssea do parafuso expansor cervical / Histomorphometric analysis of bone-screw interface of expansive cervical screw

OBJETIVO: Realizar estudo experimental para a avaliação histomorfométrica da interface óssea dos parafusos expansores utilizados no sistema de fixação anterior da coluna cervical. Métodos: Foram utilizadas no estudo cinco vértebras cervicais de ovelhas (C4), nas quais os parafusos foram inseridos. O parafuso expansor de 18,5mm de comprimento e 5,0mm de diâmetro externo (Ulrich) foi inserido em ambos os lados da véterbra C4. No lado esquerdo o parafuso era inserido sem o parafuso interno de expansão e, no lado direito, com o parafuso de expansão interna. Na porção inferior da vértebra foi confeccionado o orifício-piloto com broca de 2,5mm sem a introdução de implante. A região da vértebra contendo os parafusos e o orifício-piloto foi preparada para estudo histológico da interface dos implantes e o tecido ósseo da parede do orifício-piloto. Por meio do estudo histomorfométrico foi avaliada a densidade óssea total, a densidade óssea externa (fora da rosca do parafuso), a densidade interna (dentro da rosca do parafuso) e a medida linear de contato entre o tecido ósseo e o implante. RESULTADOS: A densidade óssea total foi maior no grupo I (parafuso expandido) em relação ao grupo III (controle). A densidade óssea externa foi maior nos grupos I (parafuso expandido) e II (não expandido) em relação ao grupo III. A densidade interna foi maior no grupo I em relação aos grupos II e III e maior no grupo II em relação ao grupo III. O contato linear foi maior no grupo I em relação aos grupos II e III. CONCLUSÃO: As alterações da estrutura do tecido ósseo ao redor dos parafusos expansores, detectadas imediatamente após a sua aplicação, forneceram subsídios para o entendimento da maior resistência ao arrancamento desses implantes, que poderiam estar relacionadas com a compactação do osso esponjoso ao redor do mesmo, proporcionando maior área de contato entre o implante e o tecido ósseo.

OBJECTIVE: To perform an experimental study for the Histomorphometric analysis of the bone interface of expansive screws used in the anterior cervical spine fixation system. METHODS: Five sheep cervical vertebrae (C4) were used in the study, in which screws were inserted. The 18.5 mm long and 5.0 mm outer diameter expansive screws (Ulrich) were inserted in both sides of the C4 vertebra. On the left side, the screw was inserted without the internal expansive screw, which was used on the right side. A pilot hole was made on the lower portion of the vertebra with a 2.5 mm bore, but no implant was introduced. The vertebral region containing the screws and the pilot hole was prepared for a histological study of the interface between implants and bone screws, and of the pilot hole wall. Histomorphometric analysis evaluated total bone density, external bone density (outside the screw thread), internal density (inside the screw thread) and the linear measurement of the contact between bone tissue and the implant. RESULTS: Total bone density was greater in group I (expansive screw) as compared to group III (control). External bone density was greater in groups I (expansive screw) and II (non-expansive screw) when compared to group III. Internal bone density was greater in group I when compared to groups II and III, and greater in group II when compared to group III. Linear contact was greater in group I when compared to groups II and III. CONCLUSION: structural changes around expansive screws detected immediately after their application provide subsidies for a better understanding of the greater tearing resistance of the implant, which could be related to the compacting of cancellous bone around the implant, while providing a larger contact area between the implant and the bone tissue.

Estudo multicêntrico do implante de stent intracoronário no Rio de Janeiro / Multicenter study of intracoronary stent implantation in Rio de Janeiro

OBJETIVO: Avaliar retrospectivamente, implante de stent (IS) em paciente com doença arterial coronária (DAC), realizado em 7 hospitais do Rio de Janeiro. Métodos: De junho/94 a dezembro/96, foram realizados 2.220 procedimentos, e desses, Analisados 783 IS em 660 (29,7 "por cento" pacientes, através da coronariografia sem subtraçäo digital. Os diversos tipos de stent, mais frequênte, usados foram: Palmaz-Schatz (40,9 "por cento"), Gianturco-Roubin (29,1 "por cento" ) e NIR (22,0 "por cento"). As indicaçöes para o IS foram: lesäo de novo, 67,9 "por cento"; lesäo reestenótica, 16,0 "por cento"; lesäo com resultado subótimo da angioplastia transluminal coronária (ATC), 8,2 "por cento"; oclusäo aguda ou provável pós-ATC, 4,9 "por cento"; oclusäo crônica, 3,0 "por cento". Todos os stents foram implantados com alta pressäo, sem controle pelo ultra-som intracoronário. A prevençäo da trombose subaguda, na maioria dos pacientes (87,8 "por cento", foi feita com o uso de ticlopidina e ácido acetil-salicílico. RESULTADOS: Evoluçä imediata: a) sucesso no IS em 770 lesöes: 98,0 "por cento" em 646 (97,9 "por cento") pacientes; b)sucesso clínico em 634 (96,0 "por cento") casos; c) complicaçöes maiores : infarto agudo do miocárdio (IAM) - 1,1 "por cento"; cirurgia de revascularizaçäo miocárdica (CRM) - 1,4 "por cento" e óbito - 0,8 "por cento"; d) complicaçöes vasculares com correçäo cirúrgico e/ou sangramento - 3,0 "por cento". Evoluçäo tardia: a) dos 399 (60,4 "por cento") pacientes acompanhados clinicamente, a coronariografia de 121 (30,3 "por cento") mostrou retorno da lesäo no IS em 79 (19,8 "por cento") casos; b) frequência de outros eventos: IAM - 1,5 "por cento"; CRM - 2,3 "por cento"; óbito - 1,0 "por cento" e outra ATC ou procedimento similar - 12,5 "por cento". CONCLUSÄO: O estudo multicêntrico mostrou que o IS na DAC pode ser realizado com segurança , alto índice de sucesso imediato, poucas complicaçöes e baixa taxa de eventos cardíacos na evolucäo tardia.

Múltiplas fístulas das artérias coronárias ao ventrículo esquerdo / Multiple Coronary Artery to Left Ventricle Fistulae

A 51 year-old man was admitted to the coronary care unit due an episode of retroestenal pain with ischaemic alterations in electrocardiogram. It was made the diagnosis of unstable angina. Cinecoronariography showed dilated coronary arteries without obstructions and multiples coronary fistulas draining into left ventricular cavity. Transesophagic echocardiogram with microbubbles injection into coronary arteries showed an important coronary steal phenomena. It is discussed the diagnosis and the treatment. This is one of the rare cases of coronary fistulas draining into the left ventricular cavity and the first one with alterations in the rest ECG.