Foraminiferal biotopes and their distribution control in Ria de Aveiro (Portugal): a multiproxy approach.

Ria de Aveiro, which is located in the centre of Portugal (40° 38' N, 8° 45' W), is a well-mixed and complex coastal lagoon that is separated from the sea by a sandy barrier and connects with the Atlantic through an artificial inlet. Tidal currents are the main factor controlling the lagoon's hydrodynamics and, to a great extent, the sedimentary dynamic. The inner lagoonal zones receive input from several rivers and experience the pressure caused by the accumulation of organic matter and pollutants (namely, trace metals) from diverse anthropic activities. This paper is the first piece of work aiming to recognize, characterize and explain the main benthic foraminiferal biotopes in Ria de Aveiro. To provide a broad overview of this kind of setting, our results are compared to those of previous published studies conducted in similar transitional environments. The research is based on an investigation of 225 sites spread throughout this ecosystem. Utilizing a statistical approach, this study analyses the details of dead benthic foraminiferal assemblages composed of 260 taxa, the texture and composition (mineralogical and geochemical) of the sediment and physicochemical data. On the basis of the results of R-mode and Q-mode cluster analyses, several different biotopes can be defined as marsh biotope/near-marsh biotope; marginal urban/marginal urban mixing biotope; inner-outer lagoon biotope or enclosed lagoon; outer lagoon biotope, mixed sub-biotope; and outer lagoon, marine sub-biotope. These biotopes are related to foraminifera assemblages and substrate type and are influenced by local currents, water depth, chemical and physicochemical conditions, river or oceanic proximity, and anthropogenic impact, as evidenced by the mapping of the six factor loadings of the principal component analysis conducted herein. Based on a similar methodology of analysis as that applied in previous studies in the Lagoon of Venice, comparable biotypes were identified in Lagoon of Aveiro.

Pulsed FCAW of Martensitic Stainless Clads onto Mild Steel: Microstructure, Hardness, and Residual Stresses.

The low carbon martensitic stainless AWS 410NiMo steel has in its chemical composition 13% chromium, 4% nickel, and 0.4% molybdenum (wt.%) and is used in turbine recovery, rotors, and high-pressure steam pump housings due to its resistance to impact at low temperatures, as well as to corrosion and cavitation. Those applications of the AWS 410NiMo steel frequently demand repair, which is performed by welding or cladding. Arc welding is a well-established technique for joining materials and presents several parameters that influence the mechanical performance of the weld bead. Although numerous welding processes exist, optimizing welding parameters for specific applications and materials is always challenging. The present work deals with a systematic study to verify the correlation between the pulsed fluxed core arc welding (FCAW) parameters, namely pulse current and frequency, welding speed, and contact tip work distance (CTWD), and the bead morphology, microstructure formation, residual stress, and hardness of the martensitic clad. The substrate used was the AISI 1020 steel, and the AWS 410NiMo steel was the filler metal for clad deposition. From the initial nine (9) samples, three (3) were selected for in-depth characterization. Lower heat input resulted in lower dilution, more elevated hardness, and lower compressive residual stresses. Therefore, the results highlight the need for selecting the proper heat input, even when using a pulsed FCAW procedure, to achieve the desired performance of the clad. In the present case, a higher heat input appears to be more advantageous owing to the lower convexity index, smooth hardness transition between fusion and heat-affected zones in addition to more elevated compressive stresses.

Adaptation and convergence in circadian-related genes in Iberian freshwater fish.

BACKGROUND: The circadian clock is a biological timing system that improves the ability of organisms to deal with environmental fluctuations. At the molecular level it consists of a network of transcription-translation feedback loops, involving genes that activate (bmal and clock - positive loop) and repress expression (cryptochrome (cry) and period (per) - negative loop). This is regulated by daily alternations of light but can also be affected by temperature. Fish, as ectothermic, depend on the environmental temperature and thus are good models to study its integration within the circadian system. Here, we studied the molecular evolution of circadian genes in four Squalius freshwater fish species, distributed across Western Iberian rivers affected by two climatic types with different environmental conditions (e.g., light and temperature). S. carolitertii and S. pyrenaicus inhabit the colder northern region under Atlantic climate type, while S. torgalensis, S. aradensis and some populations of S. pyrenaicus inhabit the warmer southern region affected by summer droughts, under Mediterranean climate type. RESULTS: We identified 16 circadian-core genes in the Squalius species using a comparative transcriptomics approach. We detected evidence of positive selection in 12 of these genes using methods based on dN/dS. Positive selection was mainly found in cry and per genes of the negative loop, with 55 putatively adaptive substitutions, 16 located on protein domains. Evidence for positive selection is predominant in southern populations affected by the Mediterranean climate type. By predicting protein features we found that changes at sites under positive selection can impact protein thermostability by changing their aliphatic index and isoelectric point. Additionally, in nine genes, the phylogenetic clustering of species that belong to different clades but inhabit southern basins with similar environmental conditions indicated evolutionary convergence. We found evidence for increased nonsynonymous substitution rate in convergent lineages, likely due to positive selection at 27 sites, mostly in cry genes. CONCLUSIONS: Our results support that temperature may be a selective pressure driving the evolution of genes involved in the circadian system. By integrating sequence-based functional protein prediction with dN/dS-based methods to detect selection we uncovered adaptive convergence in the southern populations, probably related to their similar thermal conditions.

Whole-genome analysis uncovers loss of blaZ associated with carriage isolates belonging to methicillin-resistant Staphylococcus aureus (MRSA) clone ST5-VI in Cape Verde.

OBJECTIVES: Surveillance studies for Staphylococcus aureus carriage are a primary tool to survey the prevalence of methicillin-resistant S. aureus (MRSA) in the general population, patients and healthcare workers. We have previously reported S. aureus carriage in various African countries, including Cape Verde. METHODS: Whole-genome sequences of 106 S. aureus isolates from Cape Verde were determined. RESULTS: Staphylococcus aureus carriage isolates in Cape Verde show high genetic variability, with the detection of 27 sequence types (STs) and three primary genetic clusters associated with ST152, ST15 and ST5. One transmission event with less than eight core-genome single nucleotide polymorphisms (cgSNP) differences was detected among the ST5-VI MRSA lineage. Genetic analysis confirmed the phenotypic resistance and allowed the identification of six independent events of plasmid or transposon loss associated with the deletion of blaZ in nine isolates. In the four ST5 MRSA isolates, loss of the blaZ plasmid coincided with the acquisition of SCCmec type VI and an unusual penicillin phenotype with a minimum inhibitory concentration (MIC) at the breakpoint, indicating an adaptation trend in this endemic lineage. Similar events of blaZ plasmid loss, with concomitant acquisition SCCmec elements, were detected among ST5 isolates from different geographical origins. CONCLUSION: Overall, the genome data allowed to place isolates in a phylogenetic context and to identify different blaZ gene deletions associated with plasmid or transposon loss. Genomic analysis unveiled adaptation and evolution trends, namely among emerging MRSA lineages in the country, which deserve additional consideration in the design of future infection control protocols.

A five-year retrospective study shows increasing rates of antimicrobial drug resistance in Cabo Verde for both Staphylococcus aureus and Escherichia coli.

OBJECTIVES: Data on baseline drug resistance important in informing future antimicrobial stewardship programs. So far, no data on the antimicrobial drug resistance of clinical isolates available for the African archipelago of Cabo Verde. METHODS: We performed a retrospective analysis over years (2013-17) of the drug susceptibility profiles of clinical isolates in the two main hospitals of Cabo Verde. For Escherichia coli and Staphylococcus aureus, representing 47% and 26% of all clinical isolates, the antimicrobial drug resistance profile was reported for six representative drugs. RESULTS: For E. coli we detected an increase in resistance to ampicillin, amoxicillin/clavulanic acid, ceftriaxone, ciprofloxacin and trimethoprim-and for S. aureus to methicillin, erythromycin and trimethoprim-sulfamethoxazole. This increase in both the most commonly isolated bacterial pathogens is alarm as it might compromise empirical treatment in a setting with limited access to laboratory testing. CONCLUSIONS: When compared to the published low resistance rates in carriage isolates, the more alarming situation in clinical isolates for S. aureus might encourage antimicrobial stewardship programs to reduce in hospital settings, possibly as part of the Cabo Verdean national plan against antimicrobial drug resistance.

A method for the rapid evaluation of leather biodegradability during the production phase.

Advances in technological-industrial processes have led to the development of new materials that generate different impacts on the environment when presented as waste. The application of sustainable manufacturing practices in order to improve the environmental behaviour of materials, including in the waste stage, is now an important industry responsibility. This study developed a new method for the rapid evaluation of leather biodegradability that can easily be operated by the tannery industry during the production phase. The method uses the OxiTop® system within which a solid sample is suspended in a liquid medium with no nutritional limitations at a constant temperature and stirring conditions. Ten leather samples were tested based on the existing methodology for determining aerobic biological activity (EN 16087-1: 2011), ultimate aerobic biodegradability of plastic materials in an aqueous medium (ISO 14851: 2004), and OECD 301F guidelines for testing of chemicals. The developed method has been shown to reliably distinguish (over 7 days) between samples produced using different manufacturing processes/treatments. Starch proved to be a better standard reference material for checking inoculum activity and the proper functioning of the measurement system than cellulose. Skin without treatment was shown to be a suitable reference material for defining the maximum biodegradation of leather materials. Double exponential and Gompertz mathematical models closely described the biodegradation of the tested samples. This method offers a way for industry to test and produce leather materials with higher levels of biodegradability, thus reducing the adverse environmental impacts of the final products when presented as solid waste.

Peri-implant Atypical Fractures Associated with Bisphosphonates: Should this Clinical Entity be Included in the Definition of Atypical Femoral Fracture? Case Report.

INTRODUCTION: In recent years, growing evidence of the association between bisphosphonate use and incidence of atypical femoral fractures has emerged. Cases of peri-implant fractures associated with bisphosphonate use suggest the emergence of a new clinical entity because they are not currently covered by the definition of atypical femoral fracture. CASE REPORT: We present here two rare clinical cases treated in our hospital. Two women (68 and 76 years old) with peri-implant (cephalomedullary nail) fractures and history of prolonged bisphosphonate use (11 and 14 years). Both fractures occurred after minor trauma and showed an atypical fracture pattern. One of the patients underwent revision osteosynthesis and had a 12 months follow-up. The second patient was submitted to orthopedic treatment and had a 6months follow-up. CONCLUSION: Although reports of periprosthetic fractures and peri-implant(plate and screws) have been published, references to peri-implant cephalomedullary nail fractures were not found in the literature. Here, we question if these fracture types should be included in the current definition of atypical femoral fracture, or form a new clinical entity.

Protein analysis and gene expression indicate differential vulnerability of Iberian fish species under a climate change scenario.

Current knowledge on the biological responses of freshwater fish under projected scenarios of climate change remains limited. Here, we examine differences in the protein configuration of two endemic Iberian freshwater fish species, Squalius carolitertii and the critically endangered S. torgalensis that inhabit in the Atlantic-type northern and in the Mediterranean-type southwestern regions, respectively. We performed protein structure modeling of fourteen genes linked to protein folding, energy metabolism, circadian rhythms and immune responses. Structural differences in proteins between the two species were found for HSC70, FKBP52, HIF1α and GPB1. For S. torgalensis, besides structural differences, we found higher thermostability for two proteins (HSP90 and GBP1), which can be advantageous in a warmer environment. Additionally, we investigated how these species might respond to projected scenarios of 3° climate change warming, acidification (ΔpH = -0.4), and their combined effects. Significant changes in gene expression were observed in response to all treatments, particularly under the combined warming and acidification. While S. carolitertii presented changes in gene expression for multiple proteins related to folding (hsp90aa1, hsc70, fkbp4 and stip1), only one such gene was altered in S. torgalensis (stip1). However, S. torgalensis showed a greater capacity for energy production under both the acidification and combined scenarios by increasing cs gene expression and maintaining ldha gene expression in muscle. Overall, these findings suggest that S. torgalensis is better prepared to cope with projected climate change. Worryingly, under the simulated scenarios, disturbances to circadian rhythm and immune system genes (cry1aa, per1a and gbp1) raise concerns for the persistence of both species, highlighting the need to consider multi-stressor effects when evaluating climate change impacts upon fish. This work also highlights that assessments of the potential of endangered freshwater species to cope with environmental change are crucial to help decision-makers adopt future conservation strategies.

Disruption of spermatogenesis and differential regulation of testicular estrogen receptor expression in mice after polychlorinated biphenyl exposure.

The testicular toxicity of polychlorinated biphenyls (PCBs) has been extensively studied. However, the detailed mechanism is still obscure. In the present study, male C57 mice were treated with different doses of Aroclor 1254 (a commercial PCB mixture) once every 3 days by oral gavage. After exposure to Aroclor 1254 for 50 days, the sperm count decreased in a dose-dependent manner. Cell proliferation and apoptosis are key processes regulating development of the testis, and alterations in these processes may underlie testicular dysgenesis. Our results showed that the germ cell proliferation was inhibited and the apoptosis of the germ cell was induced in a dose-dependent manner after treatment with Aroclor 1254. Although there was no significant change in serum testosterone levels and androgen receptor expression levels after treatment with different dosages of Aroclor 1254, the estradiol levels decreased and the expression of estrogen receptor (ER) ß increased in a dose-dependent manner, whereas an elevation of the expression of ERα was only observed in the 50µg/kg group. The data as a whole suggested that inhibited proliferation and induced apoptosis in germ cells, and a differential regulation of ER, may be involved in the testicular toxicity of PCBs.

Avaliações toxicológicas das alterações genotóxicas e epigenóticas induzidas por Aroclor 1254 em testí­culo, espermatozóides, fí­gado e rim de camundongos / Toxicological assessments of genotoxic and epigenetic changes induced by Aroclor 1254 in testis, sperm, liver and kidney of mice

As bifenilas policloradas (PCBs) são um grupo de compostos hidrocarbonetos halogenados aromáticos, bioacumulativos em organismos vivos e persistente no ambiente. Além da atividade disruptora endócrina, os PCBs podem aumentar os níveis de espécies reativas de oxigênio (ROS), levando ao estresse oxidativo e alteração da metilação de DNA que são fatores importantes nas etiologias da hepatotoxicidade, infertilidade masculina e doença renal. Estes agentes tóxicos podem causar disfunção mitocondrial e distúrbios que afetam a produção de ATP, ROS e morte celular, ocasionando danos à saúde humana. O presente trabalho tem como objetivo investigar possíveis alterações genotóxicas e epigenéticas causadas pelo aroclor 1254 em fígado, rim e testículo, além de verificar a indução de estresse oxidativo e disrupção dos metabólitos intermediários do ciclo de Krebs nos referidos tecidos. Camundongos machos C57/BL6 foram expostos ao Aroclor 1254 em diferentes doses (5, 50, 500 e 1000 ug/kg) por gavagem, uma vez a cada três dias, durante 50 dias. Após a exposição, os animais foram eutanasiados, os órgãos coletados e espermatozoides obtidos a partir dos epidídimos. A peroxidação lipídica em plasma e tecidos foi avaliada pela quantificação de malonaldeído (MDA) por HPLC/DAD. Os níveis de intermediários da via glicolítica, do ciclo de Krebs, de alguns nucleotídeos e aminoácidos, marcas epigenéticas (5-mC e 5-hmC) e adutos de DNA (8-oxodG e CEdG) foram quantificados por HPLC-ESI-MS/MS. A abordagem de benchmark dose (BMD) foi utilizada para a modelagem dose resposta. Após exposição, não foram observadas diferenças significativas da variação da massa corporal, e a razão do peso testicular, fígado e rim por massa corporal. No tecido hepático, foi observado aumento da peroxidação lipídica. Houve redução significativa dos níveis de ATP, ADP, razão NADP+/NADPH, piruvato, malato, fumarato e glutamato. Observou-se redução significativa dos níveis de 5-mC e 5-hmC no DNA nuclear (nDNA), enquanto não foram observadas alterações dos níveis dos adutos. Em DNA mitocondrial (mtDNA) não foram observadas alterações nas marcas epigenéticas, no entanto foi obtido aumento significativo no aduto 8-oxodG após exposição ao Aroclor 1254. No tecido renal foi observado aumento significativo de MDA. Houve aumento significativo dos níveis de lactato e malato e reduções de ATP, ADP, glutamina, NAD+. Foi observada a hipohidroximetilação do mtDNA. As marcas 5-mC de mtDNA, 5mC de nDNA e adutos de DNA nuclear e mitocondrial não apresentaram diferenças após exposição a PCBs. Nos testículos foi verificada redução significativa dos níveis de glutamato, malato, succinato, fumarato e razão NADH/NAD+, hipohidroximetilção em mtDNA e hipermetilação em nDNA. Não foram observadas alterações de 5-mC em mtDNA e 5hmC em nDNA. Não foram verificadas alterações dos níveis de MDA e adutos em nDNA. Adicionando, foi observada redução dos níveis de 5-mC em DNA global de espermatozoide. Os limites inferiores do intervalo de confiança da BMD foram estimados para que estes marcadores possam ser usados na avaliação de riscos de PCBs. Os dados obtidos apontam o Aroclor 1254 como indutor de alterações do metabolismo intermediário, das marcas epigenéticas e estresse oxidativo. Essas alterações podem afetar vias celulares, levando à morte ou transformação, e aumentando o risco de doenças

Polychlorinated biphenyls (PCBs) are a group of aromatic halogenated hydrocarbon compounds, which bioaccumulate in living organisms and is persistent in the environment. Besides their endocrine disrupting activity, PCBs may increase the levels of reactive oxygen species (ROS), leading to oxidative stress and alter DNA methylation that are important factors in the etiology of liver toxicity, male infertility, and kidney disease. These toxic agents can cause mitochondrial dysfunction and disorders that affect the production of ATP, ROS and cell death, thereby leading to health-related problems. The present work aimed at investigating possible genotoxic and epigenetic changes caused by aroclor 1254 in the liver, kidney and testis, as well as determine the induction of oxidative stress and disruption of intermediate metabolites in these tissues. Male C57/BL6 mice were exposed to Aroclor 1254 at different doses (5, 50, 500 and 1000 µg/kg) by gavage, once every three days, for 50 days. After the exposure period, the animals were euthanized, organs collected, and sperms obtained from the epididymis. Lipid peroxidation in plasma and tissues was determined by quantification of malonaldehyde (MDA) using HPLC/DAD. The levels of intermediate metabolites, epigenetic marks (5-mC and 5-hmC) and DNA adducts (8-oxodG and CEdG) were quantified by HPLC-ESI-MS/MS. The Benchmark dose approach (BMD) was used for dose response modeling. No significant differences in body weight variation, testicular, liver and kidney weight to body weight ratio were observed after exposure. However, in hepatic tissues, an increase in lipid peroxidation was observed. There were significant decreases in the intermediate metabolites including the levels of ATP, ADP, pyruvate, NADP+/NADPH ratio, malate and fumarate, as well as glutamate. Significant reduction of 5-mC and 5-hmC levels in nuclear DNA (nDNA) were observed, whereas no changes were observed in DNA adducts. The epigenetic marks in mitochondrial DNA (mtDNA) were not changed; however, a significant increase was observed in 8-oxodG adduct after exposure to Aroclor 1254. In renal tissues, data showed a significant increase in MDA, while for the intermediate metabolites, the levels of lactate and malate were significantly elevated, whereas significant reductions were recorded for ATP, ADP, glutamine, and NAD+. Hypohydroxymethylation was observed in mtDNA. The 5-mC of mtDNA, 5mC of nDNA and nuclear and mtDNA adducts did not show differences after PCBs exposure. For the testicles, significant reductions in the levels of glutamate, malate, succinate, fumarate and NADH/NAD+ ratio were observed. The PCBs also induced hypohydroxymethylation in mtDNA and hypermethylation in nDNA, but there were no changes of 5-mC in mtDNA and 5-hmC in nDNA. A reduction of nDNA adducts 8-oxodG was observed. No changes were observed in the level of MDA and DNA adducts of nDNA. However, after PCBs exposure there was a significant decrease of 5-mC in global DNA of spermatozoa. The lower bound confidence interval on BMD, which were estimated for these markers can be used in the risk assessment of PCBs. Collectively, the data obtained in this study indicate that Aroclor 1254 induces alteration of intermediate metabolites, epigenetic marks and oxidative stress. These changes can adversely affect cells and cellular pathways, therefore increase the risk of cell death or transformation