Preparation of well-dispersed chitosan/alginate hollow multilayered microcapsules for enhanced cellular internalization

Hollow multilayered capsules have shown massive potential for being used in the biomedical and biotechnology fields, in applications such as cellular internalization, intracellular trafficking, drug delivery, or tissue engineering. In particular, hollow microcapsules, developed by resorting to porous calcium carbonate sacrificial templates, natural-origin building blocks and the prominent Layer-by-Layer (LbL) technology, have attracted increasing attention owing to their key features. However, these microcapsules revealed a great tendency to aggregate, which represents a major hurdle when aiming for cellular internalization and intracellular therapeutics delivery. Herein, we report the preparation of well-dispersed polysaccharide-based hollow multilayered microcapsules by combining the LbL technique with an optimized purification process. Cationic chitosan (CHT) and anionic alginate (ALG) were chosen as the marine origin polysaccharides due to their biocompatibility and structural similarity to the extracellular matrices of living tissues. Moreover, the inexpensive and highly versatile LbL technology was used to fabricate core-shell microparticles and hollow multilayered microcapsules, with precise control over their composition and physicochemical properties, by repeating the alternate deposition of both materials. The microcapsules' synthesis procedure was optimized to extensively reduce their natural aggregation tendency, as shown by the morphological analysis monitored by advanced microscopy techniques. The well-dispersed microcapsules showed an enhanced uptake by fibroblasts, opening new perspectives for cellular internalization.publishe

Tolerance to Abiotic Factors of Microsclerotia and Mycelial Pellets From Metarhizium robertsii, and Molecular and Ultrastructural Changes During Microsclerotial Differentiation

Metarhizium spp. fungi are able to produce resistant structures termed microsclerotia, formed by compacted and melanized threads of hyphae. These propagules are tolerant to desiccation and produce infective conidia, thus they are promising candidates to use in biological control programs. In this study, we investigated the tolerance to both UV-B radiation and heat of microsclerotia of M. robertsii strain ARSEF 2575. We also adapted the liquid medium and culture conditions to obtain mycelial pellets from the same isolate in order to compare these characteristics between both types of propagules. We followed the peroxisome biogenesis and studied the oxidative stress during differentiation from conidia to microsclerotia by transmission electron microscopy after staining with a peroxidase activity marker, and by the expression pattern of genes potentially involved in these processes. We found that despite their twice smaller size, microsclerotia exhibited higher dry biomass, yield, and conidial productivity than mycelial pellets, both with and without UV-B and heat stresses.. From the sixteen genes measured, we found an induction after 96-h differentiation in the oxidative stress marker genes MrcatA, MrcatP, and Mrgpx, the peroxisome biogenesis factors Mrpex5 and Mrpex14/17, and the photoprotection genes Mrlac1, Mrlac2, and Mrlac3. We concluded that an oxidative stress scenario is induced during microsclerotia differentiation in M. robertsii, and confirmed that due to its tolerance to desiccation, heat, and UV-B, this fungal structure could be an excellent candidate for use in biological control of pests under tropical and subtropical climates where heat and UV radiation are detrimental to entomopathogenic fungi survival and persistence.Fil: Paixão, Flávia R. S.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Huarte Bonnet, Carla. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Ribeiro-Silva, Cárita de S.. Universidade Federal de Goias. Instituto de Patologia Tropical E Saude Publica.; BrasilFil: Mascarin, Gabriel M.. Empresa Brasilera de Pesquisa Agropecuaria.; BrasilFil: Fernandes, Éverton. Universidade Federal de Goias. Instituto de Patologia Tropical E Saude Publica.; BrasilFil: Pedrini, Nicolás. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; Argentin

Semi-pelagic Longline and Trammel Net Elasmobranch Catches in Southern Portugal: Catch Composition, Catch Rates and Discards

In Portugal, elasmobranch landings have decreased substantially in recent years. In this work, elasmobranch catches in semi-pelagic longlines (1997 and 1998) were compared with those in trammel nets (2000) in the Algarve, southern Portugal areas. In the semi-pelagic longline fi shery, 7 elasmobranch species represented 33.4% (2 185 specimens) of the total fi sh catches. Among the elasmobranch species, the most abundant were Galeus melastomus (63.3%), Etmopterus pusillus (21.7%) and Scyliorhinus canicula (14.2%). Most of these elasmobranchs were discarded (68.3% in total). In the trammel net fi shery, 16 different elasmobranch species represented 4.3% (597 specimens) of total fi sh catches and the most important species were Raja undulata (43.6%) and S. canicula (10.2%). The majority of the elasmobranchs caught in trammel nets had commercial value, and only 5.4% were discarded. In both fi sheries, intra-specifi c catch rates varied with depth. Length-frequency distributions for the only species with relatively high catches in both fi sheries, S. canicula, showed that, in general, trammel nets catch larger specimens and in a narrower length range than do longlines.info:eu-repo/semantics/publishedVersio

Loop bounds on non-standard neutrino interactions

We reconsider the bounds on non-standard neutrino interactions with matter which can be derived by constraining the four-charged-lepton operators induced at the loop level. We find that these bounds are model dependent. Naturalness arguments can lead to much stronger constraints than those presented in previous studies, while no completely model-independent bounds can be derived. We will illustrate how large loop-contributions to four-charged-lepton operators are induced within a particular model that realizes gauge invariant non-standard interactions and discuss conditions to avoid these bounds. These considerations mainly affect the $\mathcal O(10^{-4})$ constraint on the non-standard coupling strength \eps_{e\mu}, which is lost. The only model-independent constraints that can be derived are $\mathcal O(10^{-1})$. However, significant cancellations are required in order to saturate this bound.Comment: Minor changes, version to be published in JHEP. 17 pages, 3 Axodraw figures, REVTeX

Simulator training in gastrointestinal endoscopy - from basic training to advanced endoscopic procedures

Simulator-based gastrointestinal endoscopy training has gained acceptance over the last decades and has been extensively studied. Several types of simulators have been validated and it has been demonstrated that the use of simulators in the early training setting accelerates the learning curve in acquiring basic skills. Current GI endoscopy simulators lack the degree of realism that would be necessary to provide training to achieve full competency or to be applicable in certification. Virtual Reality and mechanical simulators are commonly used in basic flexible endoscopy training, whereas ex vivo and in vivo models are used in training the most advanced endoscopic procedures. Validated models for the training of more routine therapeutic interventions like polypectomy, EMR, stenting and haemostasis are lacking or scarce and developments in these areas should be encouraged.info:eu-repo/semantics/publishedVersio

Influence of different surface modification treatments on silk biotextiles for tissue engineering applications

Biotextile structures from silk fibroin have demonstrated to be particularly interesting for tissue engineering (TE) applications due to their high mechanical strength, interconnectivity, porosity, and ability to degrade under physiological conditions. In this work, we described several surface treatments of knitted silk fibroin (SF) scaffolds, namely sodium hydroxide (NaOH) solution, ultraviolet radiation exposure in an ozone atmosphere (UV/O3) and oxygen (O2) plasma treatment followed by acrylic acid (AAc), vinyl phosphonic acid (VPA), and vinyl sulfonic acid (VSA) immersion. The effect of these treatments on the mechanical properties of the textile constructs was evaluated by tensile tests in dry and hydrated states. Surface properties such as morphology, topography, wettability and elemental composition were also affected by the applied treatments. The in vitro biological behavior of L929 fibroblasts revealed that cells were able to adhere and spread both on the untreated and surface-modified textile constructs. The applied treatments had different effects on the scaffoldsâ surface properties, confirming that these modifications can be considered as useful techniques to modulate the surface of biomaterials according to the targeted application.Contract grant sponsor: Portuguese Foundation for Science and Technology under POCTI and/or FEDER programs under the scope of the project TISSUE2TISSUE; contract grant number: PTDC/CTM/105703/2008Contract grant sponsor: Investigator FCT program (to A.N.L.); contract grant number: IF/00411/201

Diagnostic Ability Of Barrett’s Index to Detect Dysthyroid Optic Neuropathy Using Multidetector Computed Tomography

OBJECTIVES: The objective of this study was to evaluate the ability of a muscular index (Barrett's Index), calculated with multidetector computed tomography, to detect dysthyroid optic neuropathy in patients with Graves' orbitopathy. METHODS: Thirty-six patients with Graves' orbitopathy were prospectively studied and submitted to neuro-ophthalmic evaluation and multidetector computed tomography scans of the orbits. Orbits were divided into two groups: those with and without dysthyroid optic neuropathy. Barrett's index was calculated as the percentage of the orbit occupied by muscles. Sensitivity and specificity were determined for several index values. RESULTS: Sixty-four orbits (19 with and 45 without dysthyroid optic neuropathy) met the inclusion criteria for the study. The mean Barrett's index values (± SD) were 64.47% ± 6.06% and 49.44% ± 10.94%in the groups with and without dysthyroid optic neuropathy, respectively (p<0.001). Barrett's index sensitivity ranged from 32% to 100%, and Barrett's index specificity ranged from 24% to 100%. The best combination of sensitivity and specificity was 79%/72% for BI=60% (odds ratio: 9.2). CONCLUSIONS: Barrett's Index is a useful indicator of dysthyroid optic neuropathy and may contribute to early diagnosis and treatment. Patients with a Barrett's index >60% should be carefully examined and followed for the development of dysthyroid optic neuropathy

Trypanosomiasis-Induced Th17-Like Immune Responses in Carp

Background - In mammalian vertebrates, the cytokine interleukin (IL)-12 consists of a heterodimer between p35 and p40 subunits whereas interleukin-23 is formed by a heterodimer between p19 and p40 subunits. During an immune response, the balance between IL-12 and IL-23 can depend on the nature of the pathogen associated molecular pattern (PAMP) recognized by, for example TLR2, leading to a preferential production of IL-23. IL-23 production promotes a Th17-mediated immune response characterized by the production of IL-17A/F and several chemokines, important for neutrophil recruitment and activation. For the cold blooded vertebrate common carp, only the IL-12 subunits have been described so far. Methodology/Principal Findings - Common carp is the natural host of two protozoan parasites: Trypanoplasma borreli and Trypanosoma carassii. We found that these parasites negatively affect p35 and p40a gene expression in carp. Transfection studies of HEK293 and carp macrophages show that T. carassii-derived PAMPs are agonists of carp TLR2, promoting p19 and p40c gene expression. The two protozoan parasites induce different immune responses as assessed by gene expression and histological studies. During T. carassii infections, in particular, we observed a propensity to induce p19 and p40c gene expression, suggestive of the formation of IL-23. Infections with T. borreli and T. carassii lead to an increase of IFN-¿2 gene expression whereas IL-17A/F2 gene expression was only observed during T. carasssii infections. The moderate increase in the number of splenic macrophages during T. borreli infection contrasts the marked increase in the number of splenic neutrophilic granulocytes during T. carassii infection, along with an increased gene expression of metalloproteinase-9 and chemokines. Conclusion/Significance - This is the first study that provides evidence for a Th17-like immune response in fish in response to infection with a protozoan parasit

Nutritional and functional evaluation of inula crithmoides and mesembryanthemum nodiflorum grown in different salinities for human consumption

The nutritional composition and productivity of halophytes is strongly related to the biotic/abiotic stress to which these extremophile salt tolerant plants are subjected during their cultivation cycle. In this study, two commercial halophyte species (Inula crithmoides and Mesembryanthemum nodiflorum) were cultivated at six levels of salinity using a soilless cultivation system. In this way, it was possible to understand the response mechanisms of these halophytes to salt stress. The relative productivity decreased from the salinities of 110 and 200 mmol L−1 upwards for I. crithmoides and M. nodiflorum, respectively. Nonetheless, the nutritional profile for human consumption remained balanced. In general, I. crithmoides vitamin (B1 and B6) contents were significantly higher than those of M. nodiflorum. For both species, β-carotene and lutein were induced by salinity, possibly as a response to oxidative stress. Phenolic compounds were more abundant in plants cultivated at lower salinities, while the antioxidant activity increased as a response to salt stress. Sensory characteristics were evaluated by a panel of culinary chefs showing a preference for plants grown at the salt concentration of 350 mmol L−1. In summary, salinity stress was effective in boosting important nutritional components in these species, and the soilless system promotes the sustainable and safe production of halophyte plants for human consumption.CCMAR/Multi/04326/201, MED/UIDB/05183/202, SFRH/BD/140143/2018, SFRH/BD/149398/2019info:eu-repo/semantics/publishedVersio

A biorefinery from Nannochloropsis sp. microalga – Energy and CO2 emission and economic analyses

Are microalgae a potential energy source for biofuel production? This paper presents the laboratory results from a Nannochloropsis sp. microalga biorefinery for the production of oil, high-value pigments, and biohydrogen (bioH2). The energy consumption and CO2 emissions involved in the whole process (microalgae cultivation, harvest, dewater, mill, extraction and leftover biomass fermentation) were evaluated. An economic evaluation was also performed. Oil was obtained by soxhlet (SE) and supercritical fluid extraction (SFE). The bioH2 was produced by fermentation of the leftover biomass. The oil production pathway by SE shows the lowest value of energy consumption, 177-245 MJ/MJprod, and CO2 emissions, 13–15 kgCO2/MJprod. Despite consuming and emitting c.a. 20% more than the SE pathway, the oil obtained by SFE, proved to be more economically viable, with a cost of 365€/kgoil produced and simultaneously extracting high-value pigments. The bioH2 as co-product may be advantageous in terms of product yield or profit