Combined therapy with m-TOR-dependent and -independent autophagy inducers causes neurotoxicity in a mouse model of Machado-Joseph disease

A major pathological hallmark in several neurodegenerative disorders, like polyglutamine disorders (polyQ), including Machado-Joseph disease (MJD), is the formation of protein aggregates. MJD is caused by a CAG repeat expansion in the ATXN3 gene, resulting in an abnormal protein, which is prone to misfolding and forms cytoplasmic and nuclear aggregates within neurons, ultimately inducing neurodegeneration. Treatment of proteinopathies with drugs that up-regulate autophagy has shown promising results in models of polyQ diseases. Temsirolimus (CCI-779) inhibits the mammalian target of rapamycin (m-TOR), while lithium chloride (LiCl) acts by inhibiting inositol monophosphatase, both being able to induce autophagy. We have previously shown that chronic treatment with LiCl (10.4 mg/kg) had limited effects in a transgenic MJD mouse model. Also, others have shown that CCI-779 had mild positive effects in a different mouse model of the disease. It has been suggested that the combination of mTOR-dependent and -independent autophagy inducers could be a more effective therapeutic approach. To further explore this avenue toward therapy, we treated CMVMJD135 transgenic mice with a conjugation of CCI-779 and LiCl, both at concentrations known to induce autophagy and not to be toxic. Surprisingly, this combined treatment proved to be deleterious to both wild-type (wt) and transgenic animals, failing to rescue their neurological symptoms and actually exerting neurotoxic effects. These results highlight the possible dangers of manipulating autophagy in the nervous system and suggest that a better understanding of the potential disruption in the autophagy pathway in MJD is required before successful long-term autophagy modulating therapies can be developed.Fundação para a Ciência e Tecnologia through the projects [FEDER/FCT, POCI/SAU-MMO/60412/2004], [PTDC/SAU-GMG/64076/2006]. This work was supported by Fundação para a Ciência e Tecnologi

Soybean growth and production under straw of maize, Urochloa brizantha, Conyza spp. and Digitaria insularis

Weeds can cause serious damage during soybean development, due to allelopathy, competition for water, light and nutrients. It is necessary to investigate the influence of straw, of weeds Conyza spp. and Digitaria insularis, in soybean growth, production and composition and grains. If there is influence of allelopathic compounds at the crop. The aim of this study was to evaluate the influence of straw of maize and Urochloa brizantha (A.Rich.) R.D.Webster crops and Conyza spp., D. insularis, on growth, production and composition of grains produced by soybean. Treatments consisted of the control (absence of straw), maize straw, Urochloa brizantha straw, Conyza spp. straw (500, 1000, 1500 or 2000 kg ha-1) and D. insularis straw (500, 1000, 1500 or 2000 kg ha-1). The chlorophyll index, height of plants and insertion of the first pod, stem diameter at the collar and at 5 cm from the collar, root dry mass, number of pods and grains, weight of total grains, weight of 100 grains, protein and nitrogen (N), catalase and peroxidase contents in grains were evaluated. There was no difference between treatments for plant height, first pod height and chlorophyll index, as well as for total pods and 100 grain weight and protein content, N content and peroxidase and carboxylase enzymatic activity of the grains produced. For stem diameter, a higher value was found for the treatment with maize straw compared to the control (no straw). For dry root matter, treatments without straw and with Conyza spp. straw up to 1500 kg ha-1, differed from the treatment with maize straw. Even in some respects they provided beneficial effects compared to the absence of straw, which indicates the importance of crop residues. No allelopathic effects of weeds were observed on the growth and development of soybean. Conyza spp., D. insularis, maize or U. brizantha straws do not negatively affect soybean growth, production and grain composition

Effects of conductive and non-conductive materials on the activity of a hydrogenotrophic methanogen

Several conductive materials (CM), such as carbon nanotubes (CNT), activated carbon (AC), and magnetite, have been reported to mediate interspecies electron transfer in methanogenic environments. However, CNT also accelerated methane production (MP) of pure cultures of methanogens. We hypothesize that other CM and also non-CM may affect the methanogenic activity of pure cultures. For that purpose, we incubated the hydrogenotrophic methanogen, Methanobacteriun formicicum strain DSM 1535T, with AC, zeolite (Zeo), sand and glass beads (at 0.5 g/L), and followed MP. All materials reduced lag phases preceding the MP, and the time for complete conversion of H2/CO2 to methane. The best results were obtained with Zeo, since total hydrogen conversion occurred in less than 5 days (instead of 8 days as in the control incubated without materials). Approximately 5 days with sand, and 6 days with glass beads and AC, were necessary to achieve the complete conversion. The lag phases with AC were quite short (1 day) when compared with the control assay without materials (5 days). The initial MP (determined during the first 3 days of incubation) was improved 16 times with Zeo and 11 times with AC, when compared with the cultures incubated without materials. The results show that there is not a direct relationship between conductivity and the improvement of methanogenic activity. Other physicochemical properties of the materials might be related with the beneficial effects towards methanogens.info:eu-repo/semantics/publishedVersio

Zeolite stimulates the activity of microbial enrichments converting butyrate to methane

Conductive materials have been tested as a strategy to improve methane production (MP) in anaerobic digestion (AD) processes1,2. The effect of zeolite (0.5 g/L) towards microbial enrichments converting butyrate (10 mmol/L) to methane was investigated and compared with a microbial enrichment in the absence of the material. The enrichments were initiated with granular anaerobic sludge from a brewery WWTP as inoculum. Incubations were carried out under strict anaerobic conditions (at 37 °C) and periodically transferred to fresh medium. The results showed that, after an initial adaptation period (3 transfers), the presence of zeolite significantly accelerated the total conversion of butyrate to methane, since it took approximately 30d with zeolite and around 45d without zeolite. However, both enrichment cultures after extended adaptation (more than 8 transfers) behaved similarly, degrading butyrate in approximately 15d. Nevertheless, zeolite addition to active butyrate enrichment cultures without previous contact with zeolite, slightly accelerated MP, while the highly adapted zeolite-enrichment decreased activity when incubated without zeolite. Thus, the presence of zeolite showed to stimulate the microbial activity enhancing MP from butyrate degradation. This material possess natural ion-exchange properties, absorptive capacity and could function as a support for biomass which makes its application very attractive to AD processes.info:eu-repo/semantics/publishedVersio

Pure cultures of hydrogenotrophic methanogens are affected by modified activated carbons, zeolite, sand and glass beads

The metabolism of hydrogenotrophs has been showed to be improved in the presence of carbon nanotubes, which is relevant since they are crucial microorganisms in the conversion of waste to methane1. In this study, we investigated if other materials, with different physicochemical properties, also affect the hydrogenotrophic activity of Methanobacterium formicicum. M. formicicum was incubated separately with 0.5 g/L of sand, and commercial zeolite, glass beads and activated carbon (AC0) with and without modifications on the AC0 surface. Modifications were obtained by chemical oxidation with HNO3 (AC_HNO3), H2SO4 (AC_ H2SO4) or both (AC_HNO3_ H2SO4) and thermal treatments. All materials, with exception of AC_HNO3_ H2SO4, improved the methanogenic activity. Carbon-based materials significantly reduced the lag phases preceding methane production (MP) (from approximately 5 days in the control to circa 1 day). Zeolite, sand and glass beads also reduced the lag phases but less than carbon materials (i.e., from 5 days to 1.5, 2.7 and 3.5 days, respectively). Additionally, exponential MP rates were up to 1.5 times higher in the assays with non-carbon materials. All materials tested have different physical/chemical properties including conductivities, but all stimulated the methanogenic activity. Thus, further studies are necessary to identify the mechanisms behind the underlying observations.info:eu-repo/semantics/publishedVersio

Development and application of a mechanistic nutrient-based model for precision fish farming

This manuscript describes and evaluates the FEEDNETICS model, a detailed mechanistic nutrient-based model that has been developed to be used as a data interpretation and decisionsupport tool by fish farmers, aquafeed producers, aquaculture consultants and researchers. The modelling framework comprises two main components: (i) fish model, that simulates at the individual level the fish growth, composition, and nutrient utilization, following basic physical principles and prior information on the organization and control of biochemical/metabolic processes; and (ii) farm model, that upscales all information to the population level. The model was calibrated and validated for five commercially relevant farmed fish species, i.e., gilthead seabream (Sparus aurata), European seabass (Dicentrarchus labrax), Atlantic salmon (Salmo salar), rainbow trout (Oncorhynchus mykiss), and Nile tilapia (Oreochromis niloticus), using data sets covering a wide range of rearing and feeding conditions. The results of the validation of the model for fish growth are consistent between species, presenting a mean absolute percentage error (MAPE) between 11.7 and 13.8%. Several uses cases are presented, illustrating how this tool can be used to complement experimental trial design and interpretation, and to evaluate nutritional and environmental effects at the farm level. FEEDNETICS provides a means of transforming data into useful information, thus contributing to more efficient fish farmingGrant agreement no. 818367; FEEDNETICS 4.0, funded by EUROSTARS-2 program; FEDER/ERDF, CRESC Algarve 2020 and NORTE 2020; PT-INNOVATION-0099; LA/P/0101/2020info:eu-repo/semantics/publishedVersio

Detoxification of ciprofloxacin in an anaerobic bioprocess supplemented with magnetic carbon nanotubes: contribution of adsorption and biodegradation mechanisms

In anaerobic bioreactors, the electrons produced during the oxidation of organic matter can potentially be used for the biological reduction of pharmaceuticals in wastewaters. Common electron transfer limitations benefit from the acceleration of reactions through utilization of redox mediators (RM). This work explores the potential of carbon nanomaterials (CNM) as RM on the anaerobic removal of ciprofloxacin (CIP). Pristine and tailored carbon nanotubes (CNT) were first tested for chemical reduction of CIP, and pristine CNT was found as the best material, so it was further utilized in biological anaerobic assays with anaerobic granular sludge (GS). In addition, magnetic CNT were prepared and also tested in biological assays, as they are easier to be recovered and reused. In biological tests with CNM, approximately 99% CIP removal was achieved, and the reaction rates increased ?1.5-fold relatively to the control without CNM. In these experiments, CIP adsorption onto GS and CNM was above 90%. Despite, after applying three successive cycles of CIP addition, the catalytic properties of magnetic CNT were maintained while adsorption decreased to 29 ± 3.2%, as the result of CNM overload by CIP. The results suggest the combined occurrence of different mechanisms for CIP removal: adsorption on GS and/or CNM, and biological reduction or oxidation, which can be accelerated by the presence of CNM. After biological treatment with CNM, toxicity towards Vibrio fischeri was evaluated, resulting in ? 46% detoxification of CIP solution, showing the advantages of combining biological treatment with CNM for CIP removal.This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UIDB/04469/2020 unit and BioTecNorte operation (NORTE-01-0145-FEDER-000004) funded by the European Regional Development Fund under the scope of Norte2020-Programa Operacional Regional do Norte. Ana Rita Silva holds an FCT grant SFRH/BD/131905/2017. Cátia S.N. Braga holds an FCT grant SFRH/BD/132003/2017. This work was also financially supported by: Base Funding-UIDB/50020/2020 of the Associate Laboratory LSRE LCM-funded by national funds through FCT/MCTES (PIDDAC). OSGPS acknowledges FCT fund ing under the Scientific Employment Stimulus-Institutional Call CEECINST/00049/2info:eu-repo/semantics/publishedVersio

Modulation of butyrate-degrading methanogenic communities by conductive materials

Butyrate is a volatile fatty acid commonly present in anaerobic bioreactors. Previous research showed that methane production (MP) rates from butyrate, by lake sediment microbiomes, doubled by addition of carbon nanotubes, which was accompanied by changes in the microbial community composition, with enrichment of typical fatty-acid degrading bacteria (Syntrophomonas spp.), well known to exchange electrons with methanogens via hydrogen or formate formation1. But the authors suggested that electrons exchange via conductive materials (CM) may take place instead. In our study, anaerobic butyrate-degrading enrichment cultures were developed with other CM, namely activated carbon (AC) and magnetite (Mag) at 0.5 g/L. MP started earlier in AC enrichment and complete degradation was achieved faster in Mag enrichment. Syntrophomonas spp. were enriched in all cultures (representing 60 to 80 % of the total bacterial community), but hydrogenotrophic methanogens were highly stimulated by AC (78 % of Methanomicrobiales), while the methanogenic community of Mag culture was more diverse in acetoclastic methanogens (43% of Methanosarcina and Methanosaeta). It is still unclear if the improvement on butyrate degradation is associated to the role of CM in interspecies electron transfer, but it is undoubtful that they differentially modulate the methanogenic communities towards faster MP.info:eu-repo/semantics/publishedVersio

Resolving the ancestry of Austronesian-speaking populations

There are two very different interpretations of the prehistory of Island Southeast Asia (ISEA), with genetic evidence invoked in support of both. The “out-of-Taiwan” model proposes a major Late Holocene expansion of Neolithic Austronesian speakers from Taiwan. An alternative, proposing that Late Glacial/postglacial sea-level rises triggered largely autochthonous dispersals, accounts for some otherwise enigmatic genetic patterns, but fails to explain the Austronesian language dispersal. Combining mitochondrial DNA (mtDNA), Y-chromosome and genome-wide data, we performed the most comprehensive analysis of the region to date, obtaining highly consistent results across all three systems and allowing us to reconcile the models. We infer a primarily common ancestry for Taiwan/ISEA populations established before the Neolithic, but also detected clear signals of two minor Late Holocene migrations, probably representing Neolithic input from both Mainland Southeast Asia and South China, via Taiwan. This latter may therefore have mediated the Austronesian language dispersal, implying small-scale migration and language shift rather than large-scale expansion

Differential effects of carbon-based and iron-based conductive materials in anaerobic butyrate-degrading enrichments

Introduction Conductive materials (CM) accelerate methane production (MP), probably by promoting more efficient interactions between bacteria and methanogens. This work investigates the effects of activated carbon (AC) and magnetite (Mag) in microbial enrichments degrading butyrate. Three different butyrate-degrading enrichments were developed: 1) without CM, 2) with AC, or 3) with Mag. It was also investigated if the effect of CM persisted when CM-adapted enrichments were transferred to new medium without CM, and if CM affected the activity of stable enrichments without previous contact with CM. Methodology Enrichment series were initiated with granular anaerobic sludge as inoculum, butyrate (10 mmol/L) as substrate, and CM (0.5 g/L AC or 0.5 g/L Mag), or without CM, and incubated at 37 °C, under strict anaerobic conditions. The following parameters were monitored: methane by gas chromatography; butyrate and acetate by high performance liquid chromatography; oxidation-reduction potential; pH and conductivity. RNA was extracted and taxonomic composition of the microbial communities was obtained by 16S rRNA gene sequencing. Results During the first incubations, AC-enrichment consumed hydrogen derived from butyrate degradation within 4 days, which was much faster than the enrichments with Mag and without CM, which presented lag phases (LP), preceding MP, longer than 11 and 7 days, respectively. Thus, Mag probably inhibited butyrate-degrading bacteria and/or hydrogenotrophic methanogens. Conversely, after the lag phase, Mag-enrichment was the fastest converting acetate to methane (3 times faster than in AC-enrichment), suggesting a stimulatory effect of Mag towards acetoclastic methanogens. Nevertheless, once the enrichments were adapted to the growth conditions, more efficient butyrate conversion was observed by all enrichments, with lag phases lower than 4 days, even in the control-enrichment. No significant changes on butyrate degradation were observed when highly adapted CM-enrichments were transferred to fresh medium without CM. On the other hand, when active enrichments (without previous contact with CM), were incubated with AC, it became slightly faster (0.7 times shorter LP), and with Mag were greatly inhibited (12 times longer LP). Syntrophomonas spp. represented 60 to 80 % of the total bacterial communities in all enrichments. Hydrogenotrophs were more abundant in AC-enrichment (78 % of Methanomicrobiales) and Mag-enrichment was highly enriched in acetoclastic methanogens (43 % of microorganisms assigned to Methanosaeta and Methanosarcina). Conclusions The presence of CM affects the performance of butyrate-degrading communities, with AC accelerating particularly butyrate conversion to methane (via H2/CO2) and acetate, and Mag inhibiting that first step but stimulating acetate conversion to methane.info:eu-repo/semantics/publishedVersio