Search

Myotonic dystrophy type 1 (DM1) is the most common form of muscular dystrophy in adults affecting the skeletal muscle, heart, and brain. It is caused by a CTG repeat expansion in the 3'UTR of the DMPK gene. The CUG repeats aggregates with muscle-blind proteins (MBNLs) in nuclear foci, leading to the impairment of several mRNA processing functions, including alternative splicing. The reversal from an adult to a fetal splicing profile alters or abrogates the function of a myriad of proteins, resulting in dysfunction in multiple organs. Currently, there is no treatment available for DM1 patients. Antisense oligonucleotides (ASOs) aiming to knock down DMPK expression or bind to the CUG repeats and release MBNL have been explored, with promising results in animal models. Unfortunately, a clinical trial with these drugs, while safe, failed to show improvement in patients, probably due to intrinsic limitations of ASOs. To overcome this limitation, we designed modified U7 small nuclear RNAs containing a promoter and antisense sequences targeting the 3'UTR region to promote DMPK knockdown or steric hindrance of the CUG repeats. We used patient-derived cell lines to test our approach, and the effect of the treatment was evaluated using RNA FISH combined with MBNL1 immunostaining for foci quantification and MBNL1 localization. Gene expression and splicing profiles were assessed by RNAseq and selected candidate genes were confirmed by RT-PCR. The AAV treatment reduced the foci number, released MBNL1 from the CUG foci, and shifted the splicing profile. Based on the in vitro results, we selected one vector candidate to test in vivo. We injected AAV1.U7snRNAs in the tibialis anterior (TA) and gastrocnemius (Gas) of HSAlr mice, which express CUG repeats in skeletal muscles only. Eight weeks post-injection, we submitted the animals to muscle torque and electromyography (EMG) tests and collected the muscles for further analysis. The functional tests revealed reduced mechanical myotonia (a major phenotype in DM1 muscles) and even the absence of electrical myotonia in individual muscles. RNA FISH quantification showed a reduced number and intensity of foci in TA and Gas. The reduction in foci led to improved splicing of Serca1, Mbnl1, Ldb3, and Clcn1 transcripts. In conclusion, U7snRNAs that interfere with the CUG-expanded mRNA can reverse DM1 pathology with the restoration of molecular features in vitro and in vivo, accompanied by significant amelioration of muscle physiological properties. This work will provide the basis for testing the first AAV-based gene therapy for DM1 in humans. [ABSTRACT FROM AUTHOR]

Exon skipping, a promising modality to treat Duchenne muscular dystrophy (DMD) is based on restoring the reading frame of the DMD pre-mRNA and to make truncated but functional dystrophin. Conventional exon skipping using antisense oligonucleotides has some limitations – a need of repeated injections due to less stability and limited tissue penetration in major affected tissues (e.g., heart). To overcome these, we and others used AAV.U7 antisense delivery. U7 small nuclear RNA carrying antisense sequence improves the stability and adeno-associated virus (AAV) increases tropism. This approach is currently being tested in a clinical trial for DMD exon 2 duplication. In this study, we applied this vectorized exon skipping strategy (VES) approach to a mutational hot spot in the DMD gene: the exon 44. We evaluated our lead candidate using intramuscular (IM) and systemic (IV) pre-clinical dose escalation study. Humanized DMD mice with exon 45 deletion were used. We evaluated exon skipping by RT-PCR, proteins restoration using immunostaining, muscle function using force transducer, muscle co-ordination and strength using rotarod and hang wire, and behavior test with open field apparatus. 3-months post-IM, around 85% of exon skipping was achieved, resulting in around 90% of truncated dystrophin expression and more than 50% improvement in eccentric contraction. 3-months post-IV dose escalation, around 80% of exon skipping was observed in heart and 30–60% in diaphragm, TA and gastrocnemius muscles. Dystrophin expression and improvement in force generation in dose-dependent manner was noted. Macrophage infiltration, muscle fibrosis, rotarod, hang wire and open field test revealed improvement after treatment with the highest IV dose. To conclude, our lead candidate induces efficient DMD exon 44 skipping, resulting in dystrophin production and muscle strength improvement in DMD. This AAV.U7-exon 44 skipping vector represents a promising candidate for ∼6-12% of DMD patients. [ABSTRACT FROM AUTHOR]

[Display omitted] • Six types of canned seafood were contaminated with MPs. • Average values were 0.043 ± 0.070 MPs/g (tissue) and 0.006 ± 0.015 MPs/mL (liquid). • Blue polyester fibers were the predominant MPs detected in the samples. • Octopus in tomato and tuna in olive oil presented the highest MPs contamination. • In general, seafood tissues were more contaminated than the immersion liquids. There is limited research on the occurrence of microplastics (MPs) in canned seafood. All types of canned seafood investigated in the present study were contaminated. After sample digestion in 30 % hydrogen peroxide, a total of 40 MPs were recovered. Fibers were the most common type, blue was the dominant colour, and Fourier Transform Infrared Spectroscopy (FTIR) identified polyester as the most common polymer. Considering all samples, an average of 3.5 ± 5.2 MPs/can was obtained, with octopus in tomato sauce and tuna in olive oil presenting the highest contamination (5.2 ± 7.5 MPs/can and 5.2 ± 5.1 MPs/can, respectively). Also, significant differences between the number of MPs in the seafood tissues and immersion liquids were verified. The present study demonstrates MPs occurrence in canned seafood, a potential contamination pathway for humans. More research on the different stages of the canning processing is vital for understanding MPs contamination in cans. [ABSTRACT FROM AUTHOR]

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.dsr.2008.10.008 Byline: P. Becognee (a), M. Moyano (a), C. Almeida (a), J.M. Rodriguez (b), E. Fraile-Nuez (c), A. Hernandez-Guerra (c), S. Hernandez-Leon (a) Keywords: Fish larvae; Clupeoids; Filament; Cyclonic eddy; Canary Islands Abstract: The distribution of fish larvae in relation to a filament shed from the Northwest African coastal upwelling was studied in February 2001. During the cruise, the filament was located between 27[degrees]N and 28[degrees]N, extending from the African coastal upwelling zone to the south of Fuerteventura Island (Canary Islands). This filament was trapped and remained over the quasi-permanent cyclonic eddy observed in previous studies. Almost all Sardina pilchardus, Engraulis encrasicolus and Sardinella aurita larvae caught during the cruise were associated with upwelled waters and filament structures. The sampled larval fish assemblage was composed by 12.6% of clupeoid larvae. These were distributed as follows: 73.9% were S. pilchardus, 20.7% were E. encrasicolus and 5.4% were S. aurita. Their distribution suggested that the coastal upwelling filament is a mechanism of transport from the upwelling area to oceanic waters, but its junction with the generated cyclonic eddy may not always work as a retention structure for those transported larvae, as described in previous studies. Physiological studies based on gut fluorescence and ETS activity of clupeoid larvae, as proxies for grazing and respiration, respectively, denoted a switch from pigmented food near the upwelling zone to unpigmented food toward the ocean. This pattern agrees with observed mesozooplankton feeding along an upwelling filament in previous studies. Therefore, this work confirms the close relationship between clupeoids distribution and mesoscale circulation, as well as constitutes the first assessment of the metabolic activity of those larvae in the region. Author Affiliation: (a) Biological Oceanography Laboratory, Facultad de Ciencias del Mar, Universidad de Las Palmas de G.C., Campus Universitario de Tafira, 35017 Las Palmas de Gran Canaria, Canary Islands, Spain (b) Instituto EspaA[+ or -]ol de Oceanografia, Centro Oceanografico de Gijon, Avda. Principe de Asturias 70Bis, 33212 Gijon, Asturias, Spain (c) Physics Department, Facultad de Ciencias del Mar, Universidad de Las Palmas de G.C., Campus Universitario de Tafira, 35017 Las Palmas de Gran Canaria, Canary Islands, Spain Article History: Received 27 February 2008; Revised 25 October 2008; Accepted 28 October 2008

The vertical distribution of zooplankton biomass and indices of feeding and metabolism were studied across an anticyclonic eddy of Canary Islands. The result suggested that the anticyclonic eddy enhanced both migration from deep waters and active flux.

Following a fuel leakage inside a Portuguese maritime port, we conducted parallel 30-day experiments using contaminated seawater and fuel, sampled five days after the incident. This study aimed to (i)survey the native microbial community response to the spilled fuel and (ii)evaluate the efficacy of bioremediation, both biostimulation and bioaugmentation with a lyophilized bacterial consortium (Rhodococcus erythropolis , Pseudomonas sp.), in accelerating hydrocarbon degradation. Metabarcoding analysis revealed a shift in microbial communities, with increased abundance of hydrocarbon-degraders (e.g. Alcanivorax , Thalassospira). Ninety-five hydrocarbonoclastic bacteria were isolated, including key groups from the enriched communities. The lyophilized bacteria added in bioaugmentation, enhanced the abundance of hydrocarbon-degraders over time and were recovered throughout time. Bioremediation treatments favoured biodegradation, achieving over 60 % removal of total petroleum hydrocarbons after 15 days, contrasting with natural attenuation where almost no TPH was removed. This work highlights the potential of bioremediation technologies to accelerate hydrocarbon-degrading activity, for oil spills inside ports. [Display omitted] • Spilled fuel and seawater from a port were used for enrichment experiments. • Enriched communities were dominated by Proteobacteria and Bacteroidota members. • Lyophilized bacterial consortium enhanced the abundance of HC-degraders early on. • Bioremediation treatments favoured the biodegradation process (>60 % TPH removal). [ABSTRACT FROM AUTHOR]

Abstract Estuaries are productive environments used by many fish as nursery grounds. The initial stages of fishes are highly vulnerable to (a)biotic factors, and anthropogenic pressures, influencing fish larvae assemblages along the estuary. Microplastics (MPs < 5 mm) are particularly dangerous to early life stages of fishes because their ingestion can induce gut blockage, limiting food intake or exposing organisms to contamination due to MPs capacity to absorb pollutants. Present work aimed to investigate the contamination of an urban impacted estuary (Douro estuary, NW Portugal) by MPs, and study the abundance and distribution of MPs and fish larvae in this estuary. Monthly sampling surveys were performed from December 2016 to December 2017, in nine stations along the estuary. Sub-surface planktonic horizontal trawls were performed to collect fish larvae and MPs. Planktonic samples were sorted, and fish larvae identified. MPs density was determined using a protocol optimized in our laboratory. A total of 1498 fish larvae belonging to 32 taxa were collected, with a mean density of 11.66 fish larvae 100 m−3. During the spring-summer period, it was observed the typical increase in the density and diversity of the larval assemblage. Diversity was generally low, with the high dominance of very few taxa, namely the common goby, Pomatoschistus microps. Different types of MPs were found, namely fibers, soft/hard plastic, colorful/transparent plastic, in a total of 2152 particles, with a mean density of 17.06 MPs 100 m−3. Hard MPs and fibers were the most predominant types, representing 83% of the total MPs collected. In some months the number of MPs surpassed the number of fish larvae, with an average ratio of 1.0 fish larvae:1.5 MPs. Such results are concerning, highlighting that a higher availability of MPs may facilitate their ingestion by fish and therefore increase possible impacts in these communities. Graphical abstract Unlabelled Image Highlights • First assessment of microplastics contamination in Douro estuary (Portugal) • Microplastics, including hard fragments and fibers were present over the year. • Average ratio of 1 fish larvae:1.5 microplastics • There was not a temporal or spatial overlap between fish larvae and microplastics. [ABSTRACT FROM AUTHOR]

Highlights • Removal of enrofloxacin and ceftiofur was higher than 90% in a CW fed with livestock wastewater. • Removal of enrofloxacin and ceftiofur resistance bacteria was higher than 90% for 20 one week cycles. • No significant increase of antibiotic resistance bacteria was observed in CW microcosm effluents. • CWs can be a complementary wastewater treatment when its functionality is assured. Abstract This study aimed to evaluate the potential of CWs to remove bacteria, including antibiotic resistant bacteria, and antibiotics from livestock (swine production) wastewater. Microcosms were assembled with Phragmites australis (Cav.) Trin. ex Steud and feed with wastewater spiked or not with 100 µg/L of enrofloxacin or of ceftiofur, alone or as a mixture. Wastewater (spiked or not) was treated during twenty one-week cycles. Treated wastewater was collected at weeks 1, 2, 4, 8, 14, 18 and 20 to evaluate the removal of pollutants, including nutrients, organic matter, metals and added antibiotics, and total cultivable and antibiotic resistant bacteria (enterobacteria and enterococci), which were enumerated in plate culture media. After treatment, a significant reduction in pollutants load (up to 99% depending on the parameter) was observed regardless of the presence of the veterinary antibiotics. Antibiotics were also significantly removed (removal > 90%). High removal rates were achieved for enterobacteria and enterococci bacteria during the experimental period, in the presence or absence of antibiotics (mean removals between 86 and 100%). For antibiotic resistant bacteria initially present in the influent wastewater mean removals > 90% were obtained, and no significant effect of time or of antibiotic presence was observed. This study is, to our knowledge, the first to focus on the removal of enrofloxacin and ceftiofur resistance in faecal bioindicators in CWs fed with livestock wastewater. Results confirm CWs as a green alternative to reduce the risks associated with the release of livestock effluents into the environment. [ABSTRACT FROM AUTHOR]

Abstract The present work aimed to explore the potential of autochthonous microorganisms from an urban estuary and from activated sludge of an associated wastewater treatment plant (WWTP), for biodegradation of an antidepressant drug, paroxetine, and on a cholesterol-lowering agent, bezafibrate. These compounds were chosen as representatives of extensively used pharmaceuticals. Autochthonous microorganisms from the indicated sources were exposed to the target pharmaceuticals (1 mg/L) in co-metabolism with sodium acetate (500 mg/L) along a two-weeks period, for a total of 7 two-weeks periods (here referred as cycles). Exposures were carried out in batch mode, under different incubation conditions (agitation vs. static). Removal of pharmaceuticals was monitored at the end of each cycle, by analysing the culture medium. For paroxetine, fluoride ion release was also followed as an indicator of defluorination of the molecule. The structure of the bacterial communities was analysed by ARISA (Automated rRNA Intergenic Spacer Analysis), at the beginning of the experiment and at the end of the first and the last cycles to identify substantial changes associated with the time of exposure, the incubation conditions and the presence and type of pharmaceuticals. Incubation conditions affected not only the bacterial community structure, but also the biodegradation efficiency. At the beginning of the experiment, removal of target pharmaceuticals was found to be lower under agitation than under static conditions, but at the end of the experiment, results showed high removal of the pharmaceuticals from the culture medium (>97%) under both conditions, mainly by microbiological processes. For paroxetine, adsorption and abiotic processes also had an important influence on its removal, but defluorination only occurred in the presence of microorganisms. These results highlight that autochthonous microorganisms from estuarine sediments and WWTP sludge have high ability to remove the selected pharmaceuticals with relevant implications for the development of new bioremediation tools for environmental restoration. Graphical abstract Unlabelled Image Highlights • Potential of autochthonous microorganisms for bioremediation of pharmaceuticals tested • Microorganisms from a sediment and an activated sludge degraded paroxetine and bezafibrate • Microbial community structure changed to adapt to pharmaceuticals presence • Results are relevant for development of new bioremediation tools for environmental restoration. [ABSTRACT FROM AUTHOR]

Abstract Integrated compensatory responses of physiological systems towards homeostasis are generally overlooked when it comes to analysing alterations in biochemical parameters indicative of such processes. Here an hypothesis-driven multivariate analysis accounting for interactive multibiomarker responses was used to investigate effects of long-term exposure of Carcinus maenas to Hazardous and Noxious Substances (HNS). Adult male crabs were exposed to low and high post-spill levels of acrylonitrile (ACN) or aniline (ANL) for 21d. Bioaccumulation, feeding behaviour, and biomarkers related to mode-of-action (MoA) (detoxification, neurotransmission and energy production) were evaluated over time. Distinct temporal patterns of response to low and high exposure concentrations were depicted, with a main set of interactive multibiomarker predictors identified for each HNS (five for ACN and three for ANL), useful to follow coupled evolvement of biomarker responses. ACN caused peripheral neurotoxic effects coupled with enhanced biotransformation and significant oxidative damage particularly relevant in gills. ANL elicited alterations in central neurotransmission affecting ventilation coupled with very low levels of oxidative damage in gills. Results indicate chronic toxicity data are determinant to improve HNS hazard assessment if the aim is to obtain reliable risk calculations, and develop effective predictive models avoiding overestimation but sufficiently protective. Accounting for multibiomarker interactions brought otherwise overlooked information about C. maenas responses and MoA of ACN and ANL. Graphical abstract Image 1 Highlights • Multibiomarker interactions (MbI) were used to investigate long-term effects of two HNS. • Acrylonitrile caused peripheral neurotoxicity with biotransformation and oxidative stress. • Aniline decreased central neurotransmission coupled with low levels of oxidative damage. • Chronic toxicity data are crucial to improve HNS hazard assessment and risk prediction. • MbI brought otherwise overlooked information on modes-of-action of HNS investigated. Accounting for biomarker interactions drastically improves the quality of the model to diagnose toxic effects. [ABSTRACT FROM AUTHOR]

Coastal ecosystem services (ES) can be significantly altered by marine litter, because of its multiple economic and social impacts at different scales. However, current scientific evidence of the perceived impacts of marine litter in the different ES is still scarce in Galicia (Spain). The aim of this study was to evaluate the main perceived effects of marine litter on ES and coastal communities. By conducting 360 individual interviews to citizens from the general public of the Rías Baixas coastal region, we analyzed societal perceptions about the importance of ecosystems for human well-being and the magnitude and consequences of marine litter on coastal ecosystems. Our results indicated that the surveyed population are highly aware of the consequences of marine litter to the detriment of the ES provided by coastal areas (81% considered as of high impact), regardless of the socioeconomic characteristics (gender, age, house composition, education, residence, income). The results also suggest that supporting and regulating ES are scored as very important by the interviewees, while women are more concerned than men. The negative impact on human wellbeing was perceived as very important by citizens. The fisheries sector was perceived as the least responsible for pollution by marine litter presence (38% considered as a little responsible, and 16% as very little responsible), with a wide awareness of the role of citizens in both the cause and the solution to tackle marine litter. Citizens might play a key role in the mitigation of marine litter to collectively demand and promote active management measures, which are able to generate a positive impact on ecosystems and on the main socioeconomic variables in the long term. [ABSTRACT FROM AUTHOR]