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Lemnaceae taxonomy is challenged by the particular morphology of these tiny free-floating angiosperms. Although molecular taxonomy has helped clarify the phylogenetic history of this family, some inconsistency with morphological data leads to frequent misclassifications in the genus Lemna. Recently, the finding that Lemna japonica is an interspecific hybrid between Lemna minor and Lemna turionifera provided a clear explanation for one such taxonomic question. Here we demonstrated that L. minor is also capable of hybridizing with Lemna gibba, generating a cryptic but widespread taxon in the Mediterranean area. The nothotaxon Lemna ×mediterranea is described and compared with clones of the putative parental species L. minor and L. gibba. Genetic analysis by nuclear and plastid markers, as well as genome size measurement, revealed that two different cytotypes, diploid and triploid, originated by at least two independent hybridization events. Despite high overall similarity, morphometrical, physiological, and biochemical analyses showed an intermediate position of L. ×mediterranea between its parental species in most qualitative and quantitative characters, and also separation of the two hybrid cytotypes by some criteria. These data provide evidence that hybridization and polyploidization, driving forces of terrestrial plant evolution, contribute to duckweed genetic diversity and may have shaped the phylogenetic history of these mainly asexual, aquatic plants.
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Araceae , Hibridização Genética , Filogenia , Araceae/genética , Variação Genética , Poliploidia , Genoma de Planta , BiodiversidadeRESUMO
Amyotrophic Lateral Sclerosis (ALS) is considered the prototype of motor neuron disease, characterized by motor neuron loss and muscle waste. A well-established pathogenic hallmark of ALS is mitochondrial failure, leading to bioenergetic deficits. So far, pharmacological interventions for the disease have proven ineffective. Trimetazidine (TMZ) is described as a metabolic modulator acting on different cellular pathways. Its efficacy in enhancing muscular and cardiovascular performance has been widely described, although its molecular target remains elusive. We addressed the molecular mechanisms underlying TMZ action on neuronal experimental paradigms. To this aim, we treated murine SOD1G93A-model-derived primary cultures of cortical and spinal enriched motor neurons, as well as a murine motor-neuron-like cell line overexpressing SOD1G93A, with TMZ. We first characterized the bioenergetic profile of the cell cultures, demonstrating significant mitochondrial dysfunction that is reversed by acute TMZ treatments. We then investigated the effect of TMZ in promoting autophagy processes and its impact on mitochondrial morphology. Finally, we demonstrated the effectiveness of TMZ in terms of the mitochondrial functionality of ALS-rpatient-derived peripheral blood mononuclear cells (PBMCs). In summary, our results emphasize the concept that targeting mitochondrial dysfunction may represent an effective therapeutic strategy for ALS. The findings demonstrate that TMZ enhances mitochondrial performance in motor neuron cells by activating autophagy processes, particularly mitophagy. Although further investigations are needed to elucidate the precise molecular pathways involved, these results hold critical implications for the development of more effective and specific derivatives of TMZ for ALS treatment.
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Esclerose Lateral Amiotrófica , Doenças Mitocondriais , Trimetazidina , Camundongos , Animais , Humanos , Esclerose Lateral Amiotrófica/metabolismo , Superóxido Dismutase-1/metabolismo , Trimetazidina/farmacologia , Trimetazidina/uso terapêutico , Camundongos Transgênicos , Leucócitos Mononucleares/metabolismo , Superóxido Dismutase/metabolismo , Autofagia , Modelos Animais de DoençasRESUMO
Telomerase reverse transcriptase (TERT) is the catalytic subunit of telomerase holoenzyme, which adds telomeric DNA repeats on chromosome ends to counteract telomere shortening. In addition, there is evidence of TERT non-canonical functions, among which is an antioxidant role. In order to better investigate this role, we tested the response to X-rays and H2O2 treatment in hTERT-overexpressing human fibroblasts (HF-TERT). We observed in HF-TERT a reduced induction of reactive oxygen species and an increased expression of the proteins involved in the antioxidant defense. Therefore, we also tested a possible role of TERT inside mitochondria. We confirmed TERT mitochondrial localization, which increases after oxidative stress (OS) induced by H2O2 treatment. We next evaluated some mitochondrial markers. The basal mitochondria quantity appeared reduced in HF-TERT compared to normal fibroblasts and an additional reduction was observed after OS; nevertheless, the mitochondrial membrane potential and morphology were better conserved in HF-TERT. Our results suggest a protective function of TERT against OS, also preserving mitochondrial functionality.
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Antioxidantes , Telomerase , Humanos , Antioxidantes/metabolismo , Peróxido de Hidrogênio/metabolismo , Mitocôndrias/metabolismo , Estresse Oxidativo , Telomerase/metabolismoRESUMO
Breast cancer is the first leading tumor in women in terms of incidence worldwide. Seventy percent of cases are estrogen receptor (ER) α-positive. In these malignancies, 17ß-estradiol (E2) via ERα increases the levels of neuroglobin (NGB), a compensatory protein that protects cancer cells from stress-induced apoptosis, including chemotherapeutic drug treatment. Our previous data indicate that resveratrol (RSV), a plant-derived polyphenol, prevents E2/ERα-induced NGB accumulation in this cellular context, making E2-dependent breast cancer cells more prone to apoptosis. Unfortunately, RSV is readily metabolized, thus preventing its effectiveness. Here, four different RSV analogs have been developed, and their effect on the ERα/NGB pathway has been compared with RSV conjugated with highly hydrophilic gold nanoparticles as prodrug to evaluate if RSV derivatives maintain the breast cancer cells' susceptibility to the chemotherapeutic drug paclitaxel as the original compound. Results demonstrate that RSV conjugation with gold nanoparticles increases RSV efficacy, with respect to RSV analogues, reducing NGB levels and enhancing the pro-apoptotic action of paclitaxel, even preventing the anti-apoptotic action exerted by E2 treatment on these cells. Overall, RSV conjugation with gold nanoparticles makes this complex a promising agent for medical application in breast cancer treatment.
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Neoplasias da Mama , Nanopartículas Metálicas , Pró-Fármacos , Feminino , Humanos , Neuroglobina/farmacologia , Neoplasias da Mama/metabolismo , Resveratrol/farmacologia , Resveratrol/uso terapêutico , Receptor alfa de Estrogênio/metabolismo , Pró-Fármacos/farmacologia , Pró-Fármacos/uso terapêutico , Globinas/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Ouro/farmacologia , Estradiol/farmacologia , Paclitaxel/farmacologia , Paclitaxel/uso terapêutico , Linhagem Celular Tumoral , Apoptose , Estrogênios/farmacologiaRESUMO
BACKGROUND: Meloidae (blister beetles) are known to synthetize cantharidin (CA), a toxic and defensive terpene mainly stored in male accessory glands (MAG) and emitted outward through reflex-bleeding. Recent progresses in understanding CA biosynthesis and production organ(s) in Meloidae have been made, but the way in which self-protection is achieved from the hazardous accumulation and release of CA in blister beetles has been experimentally neglected. To provide hints on this pending question, a comparative de novo assembly transcriptomic approach was performed by targeting two tissues where CA is largely accumulated and regularly circulates in Meloidae: the male reproductive tract (MRT) and the haemolymph. Differential gene expression profiles in these tissues were examined in two blister beetle species, Lydus trimaculatus (Fabricius, 1775) (tribe Lyttini) and Mylabris variabilis (Pallas, 1781) (tribe Mylabrini). Upregulated transcripts were compared between the two species to identify conserved genes possibly involved in CA detoxification and transport. RESULTS: Based on our results, we hypothesize that, to avoid auto-intoxication, ABC, MFS or other solute transporters might sequester purported glycosylated CA precursors into MAG, and lipocalins could bind CA and mitigate its reactivity when released into the haemolymph during the autohaemorrhaging response. We also found an over-representation in haemolymph of protein-domains related to coagulation and integument repairing mechanisms that likely reflects the need to limit fluid loss during reflex-bleeding. CONCLUSIONS: The de novo assembled transcriptomes of L. trimaculatus and M. variabilis here provided represent valuable genetic resources to further explore the mechanisms employed to cope with toxicity of CA in blister beetle tissues. These, if revealed, might help conceiving safe and effective drug-delivery approaches to enhance the use of CA in medicine.
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Cantaridina , Besouros , Animais , Cantaridina/toxicidade , Besouros/genética , Genitália Masculina , Hemolinfa , Masculino , TranscriptomaRESUMO
BACKGROUND: The osteoconductive properties of bone grafting materials represent one area of research for the management of bony defects found in the fields of periodontology and oral surgery. From a physico-chemical aspect, the wettability of the graft has been demonstrated to be one of the most important factors for new bone formation. It is also well-known that argon plasma treatment (PAT) and ultraviolet irradiation (UV) may increase the surface wettability and, consequently, improve the regenerative potential of the bone grafts. Therefore, the aim of the present in vitro study was to evaluate the effect of PAT and UV treatment on the osteoconductive potential of various bone grafts. MATERIALS AND METHODS: The following four frequently used bone grafts were selected for this study: synthetic hydroxyapatite (Mg-HA), biphasic calcium phosphate (BCP), cancellous and cortical xenogenic bone matrices (CaBM, CoBM). Sixty-six serially numbered disks 10 mm in diameter were used for each graft material and randomly assigned to the following three groups: test 1 (PAT), test 2 (UV), and control (no treatment). Six samples underwent topographic analysis using SEM pre- and post-treatments to evaluate changes in surface topography/characteristics. Additionally, cell adhesion and cell proliferation were evaluated at 2 and 72 h respectively following incubation in a three-dimensional culture system utilizing a bioreactor. Furthermore, the effects of PAT and UV on immune cells were assessed by measuring the viability of human macrophages at 24 h. RESULTS: The topographic analysis showed different initial morphologies of the commercial biomaterials (e.g., Mg-HA and BCP showed flat morphology; BM samples were extremely porous with high roughness). The surface analysis following experimental treatments did not demonstrate topographical difference when compared with controls. Investigation of cells demonstrated that PAT treatment significantly increased cell adhesion of all 4 evaluated bone substitutes, whereas UV failed to show any statistically significant differences. The viability test revealed no differences in terms of macrophage adhesion on any of the tested surfaces. CONCLUSION: Within their limitations, the present results suggest that treatment of various bone grafting materials with PAT appears to enhance the osteoconductivity of bone substitutes in the early stage by improving osteoblast adhesion without concomitantly affecting macrophage viability. CLINICAL RELEVANCE: Treatment of bone grafts with PAT appears to result in faster osseointegration of the bone grafting materials and may thus favorably influence bone regeneration.
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Regeneração Óssea , Substitutos Ósseos , Argônio , Materiais Biocompatíveis , Transplante Ósseo , Durapatita , Humanos , Gases em PlasmaRESUMO
BACKGROUND: Attaining an effective mucosal attachment to the transmucosal part of the implant could protect the peri-implant bone. AIM: To evaluate if chair side surface treatments (plasma of Argon and ultraviolet light) may affect fibroblast adhesion on different titanium surfaces designed for soft tissue healing. METHODS: Grade 5 titanium discs with four different surface topographies were subdivided into 3 groups: argon-plasma; ultraviolet light, and no treatment. Cell morphology and adhesion tests were performed at 20 min, 24 h, and 72 h. RESULTS: Qualitative observation of the surfaces performed at the SEM was in accordance with the anticipated features. Roughness values ranged from smooth (MAC Sa = 0.2) to very rough (XA Sa = 21). At 20 min, all the untreated surfaces presented hemispherical cells with reduced filopodia, while the cells on treated samples were more spread with broad lamellipodia. However, these differences in spreading behavior disappeared at 24 h and 72 h. Argon-plasma, but not UV, significantly increased the number of fibroblasts independently of the surface type but only at 20 min. Statistically, there was no surface in combination with a treatment that favored a greater cellular adhesion. CONCLUSIONS: Data showed potential biological benefits of treating implant abutment surfaces with the plasma of argon in relation to early-stage cell adhesion.
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Argônio/farmacologia , Fibroblastos/citologia , Titânio/química , Adesão Celular , Proliferação de Células/fisiologia , Células Cultivadas , Fibroblastos/efeitos dos fármacos , Humanos , Técnicas In Vitro , Propriedades de SuperfícieRESUMO
The main purpose of this study was to explain the internal fine structure of potential antennal thermo- and hygroreceptive sensilla, their innervation specifics, and responses of the sensory neurons to thermal and humidity stimuli in an elaterid beetle using focused ion beam scanning electron microscopy and electrophysiology, respectively. Several essential, high temperature induced turning points in the locomotion were determined using automated video tracking. Our results showed that the sensilla under study, morphologically, are identical to the dome-shaped sensilla (DSS) of carabids. A cold-hot neuron and two bimodal hygro-thermoreceptor neurons, the moist-hot and dry-hot neuron, innervate them. Above 25-30⯰C, all the three neurons, at different threshold temperatures, switch from regular spiking to temperature dependent spike bursting. The percentage of bursty DSS neurons on the antenna increases with temperature increase suggesting that this parameter of the neurons may encode noxious heat in a graded manner. Thus, we show that besides carabid beetles, elaterids are another large group of insects with this ability. The threshold temperature of the beetles for onset of elevated locomotor activity (OELA) was lower by 11.9⯰C compared to that of critical thermal maximum (39.4⯰C). Total paralysis occurred at 41.8⯰C. The threshold temperatures for spike bursting of the sensory neurons in DSS and OELA of the beetles coincide suggesting that probably the spike bursts are responsible for encoding noxious heat when confronted. In behavioural thermoregulation, spike bursting DSS neurons serve as a fast and firm three-fold early warning system for the beetles to avoid overheating and death.
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Potenciais de Ação , Neurônios/fisiologia , Sensilas/fisiologia , Termorreceptores/fisiologia , Animais , Besouros , Temperatura Alta , Locomoção , Neurônios/ultraestrutura , Nociceptores/fisiologia , Nociceptores/ultraestrutura , Sensilas/inervação , Sensilas/ultraestrutura , Termorreceptores/ultraestruturaRESUMO
Despite thermosensation being crucial in effective thermoregulation behaviour, it is poorly studied in insects. Very little is known about encoding of noxious high temperatures by peripheral thermoreceptor neurons. In carabids, thermo- and hygrosensitive neurons innervate antennal dome-shaped sensilla (DSS). In this study, we demonstrate that several essential fine structural features of dendritic outer segments of the sensory neurons in the DSS and the classical model of insect thermo- and hygrosensitive sensilla differ fundamentally. Here, we show that spike bursts produced by the bimodal dry neurons in the antennal DSS may contribute to the sensation of noxious heat in P. oblongopunctatus. Our electrophysiological experiments showed that, at temperatures above 25 °C, these neurons switch from humidity-dependent regular spiking to temperature-dependent spike bursting. Five out of seven measured parameters of the bursty spike trains, the percentage of bursty dry neurons, the CV of ISIs in a spike train, the percentage of bursty spikes, the number of spikes in a burst and the ISIs in a burst, are unambiguously dependent on temperature and thus may precisely encode both noxious high steady temperatures up to 45 °C as well as rapid step-changes in it. The cold neuron starts to produce temperature-dependent spike bursts at temperatures above 30-35 °C. Thus, the two neurons encode different but largely overlapping ranges in noxious heat. The extent of dendritic branching and lamellation of the neurons largely varies in different DSS, which might be the structural basis for their variation in threshold temperatures for spike bursting.
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Potenciais de Ação/fisiologia , Antenas de Artrópodes/fisiologia , Besouros/fisiologia , Temperatura Alta , Células Receptoras Sensoriais/fisiologia , Animais , Antenas de Artrópodes/anatomia & histologia , Antenas de Artrópodes/ultraestrutura , Temperatura Baixa , Sensilas/anatomia & histologia , Sensilas/fisiologia , Sensilas/ultraestruturaRESUMO
Riboflavin Transporter Deficiency (RTD) is a rare genetic, childhood-onset disease. This pathology has a relevant neurological involvement, being characterized by motor symptoms, ponto-bulbar paralysis and sensorineural deafness. Such clinical presentation is associated with muscle weakness and motor neuron (MN) degeneration, so that RTD is considered part of the MN disease spectrum. Based on previous findings demonstrating energy dysmetabolism and mitochondrial impairment in RTD induced Pluripotent Stem cells (iPSCs) and iPSC-derived MNs, here we address the involvement of intrinsic apoptotic pathways in disease pathogenesis using these patient-specific in vitro models by combined ultrastructural and confocal analyses. We show impaired neuronal survival of RTD iPSCs and MNs. Focused Ion Beam/Scanning Electron Microscopy (FIB/SEM) documents severe alterations in patients' cells, including deranged mitochondrial ultrastructure, and altered plasma membrane and nuclear organization. Occurrence of aberrantly activated apoptosis is confirmed by immunofluorescence and TUNEL assays. Overall, our work provides evidence of a role played by mitochondrial dysfunction in RTD, and identifies neuronal apoptosis as a contributing event in disease pathogenesis, indicating intrinsic apoptosis pathways as possible relevant targets for more effective therapeutical approaches.
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OBJECTIVES: This in vitro study aimed to investigate the impact of bicarbonate air-abrasive powders and ultrasonic scaling with stainless steel tips on the micro- and nanotopography and roughness of three different implant-abutment junction titanium surfaces. MATERIALS AND METHODS: Three types of sterile and decontaminated titanium surfaces (RS, UTM, XA) were used for analysis. Nine disks per surface type were subjected to micro- and nanotopography analysis, scanning electron microscopy (SEM), roughness analysis, and fibroblast cultivation. Ultrasonic debridement and air polishing were performed on the surfaces. Human dermal fibroblasts were cultured on the surfaces for 5 days. STATISTICAL ANALYSIS: Data analysis adhered to ISO 25178 standards for surface texture assessment. SEM micrographs were used to reconstruct areas for extracting roughness parameters. Excel and Mex 6.0 software were utilized for quantitative and stereoscopic analysis. RESULTS: The study found varying effects on surface roughness posttreatment. RS Disco samples exhibited higher surface roughness compared with UTM and XA samples, both in average and nanoscale roughness. Decontamination led to increased surface roughness for all samples, particularly RS Disco. Fibroblast growth tests revealed enhanced cell network formation on decontaminated discs, possibly due to increased nanoscale roughness or the presence of bicarbonate salts. CONCLUSION: The study underscores the complex interplay between surface topography, microbial biofilm, and treatment efficacy in peri-implant disease management. While smoother surfaces may resist biofilm accumulation, increased nanoscale roughness postdecontamination can enhance fibroblast attachment and soft tissue integration. This dichotomy highlights the need for tailored treatment protocols that consider material-specific factors, emphasizing that successful implant therapy should balance microbial control with conducive surface characteristics for long-term osseointegration and soft tissue stability.
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Some Pyrochroidae species are known as "canthariphilous" for their attraction to cantharidin (CTD), a toxic terpene with anti-predatory effects, produced in nature by only two beetle families (Meloidae and Oedemeridae). It has been demonstrated that males of Neopyrochroa flabellata ingesting CTD are positively selected by females. Indeed, the compound is re-emitted from a glandular cranial apparatus as secretions that are licked up by females during courtship behaviour, inducing copulation. Herein, we provide the first description of the glands associated to the cranial apparatus of male Pyrochroinae using the European species Pyrochroa coccinea as a model. Morphological analyses show that the cranial apparatus consists of a concave pit lined with short setae retaining secretions emitted through numerous glandular pores. Ultrastructural investigations reveal the presence of two different class 3 glands (Gl.A and Gl.B), intermixed at the level of the pit but exhibiting distinct features. Gl.A are mainly characterised by short conducting canals, rounded nuclei and electrondense vesicles while Gl.B are characterised by long conducting canals, irregular nuclei, vesicles containing a particulate substance and a multifolded plasma membrane. Observations of sexual behaviour are also reported for P. coccinea and compared to N. flabellata, confirming the involvement of cranial apparatus secretions in courtship behaviour.
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Besouros , Humanos , Feminino , Masculino , Animais , Besouros/anatomia & histologia , Copulação , Comportamento Sexual , Cantaridina/metabolismo , TerpenosRESUMO
Pseudomonas aeruginosa is the most common pathogen infecting cystic fibrosis (CF) lungs, causing acute and chronic infections. Intrinsic and acquired antibiotic resistance allow P. aeruginosa to colonize and persist despite antibiotic treatment, making new therapeutic approaches necessary. Combining high-throughput screening and drug repurposing is an effective way to develop new therapeutic uses for drugs. This study screened a drug library of 3,386 drugs, mostly FDA approved, to identify antimicrobials against P. aeruginosa under physicochemical conditions relevant to CF-infected lungs. Based on the antibacterial activity, assessed spectrophotometrically against the prototype RP73 strain and 10 other CF virulent strains, and the toxic potential evaluated toward CF IB3-1 bronchial epithelial cells, five potential hits were selected for further analysis: the anti-inflammatory and antioxidant ebselen, the anticancer drugs tirapazamine, carmofur, and 5-fluorouracil, and the antifungal tavaborole. A time-kill assay showed that ebselen has the potential to cause rapid and dose-dependent bactericidal activity. The antibiofilm activity was evaluated by viable cell count and crystal violet assays, revealing carmofur and 5-fluorouracil as the most active drugs in preventing biofilm formation regardless of the concentration. In contrast, tirapazamine and tavaborole were the only drugs actively dispersing preformed biofilms. Tavaborole was the most active drug against CF pathogens other than P. aeruginosa, especially against Burkholderia cepacia and Acinetobacter baumannii, while carmofur, ebselen, and tirapazamine were particularly active against Staphylococcus aureus and B. cepacia. Electron microscopy and propidium iodide uptake assay revealed that ebselen, carmofur, and tirapazamine significantly damage cell membranes, with leakage and cytoplasm loss, by increasing membrane permeability. IMPORTANCE Antibiotic resistance makes it urgent to design new strategies for treating pulmonary infections in CF patients. The repurposing approach accelerates drug discovery and development, as the drugs' general pharmacological, pharmacokinetic, and toxicological properties are already well known. In the present study, for the first time, a high-throughput compound library screening was performed under experimental conditions relevant to CF-infected lungs. Among 3,386 drugs screened, the clinically used drugs from outside infection treatment ebselen, tirapazamine, carmofur, 5-fluorouracil, and tavaborole showed, although to different extents, anti-P. aeruginosa activity against planktonic and biofilm cells and broad-spectrum activity against other CF pathogens at concentrations not toxic to bronchial epithelial cells. The mode-of-action studies revealed ebselen, carmofur, and tirapazamine targeted the cell membrane, increasing its permeability with subsequent cell lysis. These drugs are strong candidates for repurposing for treating CF lung P. aeruginosa infections.
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Fibrose Cística , Infecções por Pseudomonas , Humanos , Pseudomonas aeruginosa , Fibrose Cística/microbiologia , Ensaios de Triagem em Larga Escala , Reposicionamento de Medicamentos , Tirapazamina/farmacologia , Tirapazamina/uso terapêutico , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Fluoruracila , Biofilmes , Infecções por Pseudomonas/microbiologiaRESUMO
Contamination by microplastics (0.1 µm-5 mm plastic fragments) is currently one of the major threats to the conservation of aquatic and terrestrial ecosystems worldwide. Growing awareness on this issue has led to an increase in studies on the effects of microplastics on freshwater organisms, although there are still few investigations on possible transfer of this contaminant along water trophic chains from producers to primary consumers. In this study, aquatic herbivorous larvae of the moth Cataclysta lemnata were fed on microplastic-free (control) and contaminated (MPs treatment) Lemna minuta fronds. For treatments, Lemna fronds were grown in mineral water enriched with fluorescent microparticles of poly(styrene-co-methyl methacrylate) (MPs, 100 mg/L) and then fed to the larvae as a food source. Microplastics effects on larvae were tested at 0, 7, 14 and 21 days of exposure, corresponding to sensitive phases of the insect life cycle. Contaminant impact was assessed based on some parameters such as viability, larva body size/weight, feeding alterations and regularity of the insect life cycle. Using scanning electron and fluorescence microscopy, the presence of microplastics in the larvae was verified. The finding of fluorescent microplastics in both the intestinal lumen and excrement samples showed that larvae ingested contaminated Lemna fronds. In addition, larvae fed contaminated fronds were strongly affected by the presence of microplastic contaminant over time, showing high mortality (90 %) and total inability to complete the life cycle after 21 days by failing to reach the winged adult phase. In control tests, survival rates were higher than in treatments, and 50 % of the larvae managed to pupate and emerge as moths, reaching the adult phase. The results show that there was a trophic transfer of microplastics from producer to primary consumer along a freshwater food chain, generating negative effects on the life cycle of this aquatic herbivore.
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Araceae , Lepidópteros , Poluentes Químicos da Água , Animais , Microplásticos , Plásticos , Cadeia Alimentar , Ecossistema , Poluentes Químicos da Água/análise , Larva , Estágios do Ciclo de Vida , Água DoceRESUMO
Ecosystems around the world are experiencing a major environmental impact from microplastic particles (MPs 0.1 µm-1 mm). Water, sediments, and aquatic biota show the widespread presence of this pollutant. However, MPs are rarely used in laboratory studies as they are scarcely available for purchase or expensive, especially if one wishes to trace the particle with a dye or fluorescent. Furthermore, existing preparation techniques have limited application in biological studies. In this work, we propose a new, easy, and cheap way to prepare fluorescent MPs. The protocol is based on the osmosis method in order to obtain spherical polymeric particles of P(S-co-MMA), with 0.7-9 micron diameter, made fluorescent because dye-doped with rhodamine B isothiocyanate (RITC) or fluorescein isothiocyanate (FITC). The dye loading was studied and optimized, and the MPs-dye conjugates were characterized by UV-vis FTIR and XPS spectrometry and scanning electron microscopy (SEM). Furthermore, preliminary tests on aquatic organisms demonstrated the possible use of these fluorescent MPs in bioimaging studies, showing their absorption/adsorption by duckweeds (Lemna minuta) and insect larvae (Cataclysta lemnata).
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The concentration of nanoplastics (NPs) is expected to increase in aquatic environments thus potentially threatening freshwater organisms through interactions with plastic particles that variously float, circulate in the water column or sink into the benthos. Studies into the mechanisms of any NP effects are still scarce, particularly with respect to the regenerative ability of biota for which there is no recognised model organism. The present study therefore aimed to investigate behavioural and regeneration responses of the freshwater planarian Girardia tigrina after 10 days exposed to along a gradient 0.01-10 mg/L of poly (styrene-co-methyl methacrylate) NPs (â¼426 ± 175 nm). Exposure to NPs induced a significant reduction in planarian feeding rate even at low concentrations (LOEC of 0.01 mg/L), while head regeneration was delayed in a clear dose response way (LOEC of 0.1 mg/L for blastema length). Planaria locomotion assessed was not affected. Our results highlight the potential adverse effects of exposure to poly (styrene-co-methyl methacrylate) NPs and show that feeding behaviour and regeneration of a freshwater benthic organism can be indicators of the resulting toxicity. Planarians are becoming widely used model organisms in ecotoxicology and can help to address potential effects of plastic polymers on regeneration.
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Planárias , Poluentes Químicos da Água , Animais , Microplásticos , Poluentes Químicos da Água/toxicidade , Água Doce , Organismos Aquáticos , Metacrilatos/farmacologia , EstirenosRESUMO
(1) Background: The aim of this study is to understand from a microscopic point of view whether bicarbonate air-abrasive powders associated with ultrasonic instruments can decontaminate nine different surfaces used for the abutment/implant junction. Fibroblast growth was carried out on decontaminated surface in order to understand if there are significative differences in terms of biocompatibility. (2) Methods: After taking samples of patient plaque, nine different surfaces were contaminated and analyzed by SEM, then their wettability was evaluated. Fibroblasts were cultured on the decontaminated surfaces to understand their ability to establish a connective tissue seal after decontamination. The results were analyzed from a statistical point of view to hypothesize a mathematical model capable of explaining the properties of the surfaces. (3) Results: A negative correlation between roughness and contamination has been demonstrated, whereas a weak correlation was observed between wettability and decontamination capacity. All surfaces were topographically damaged after the decontamination treatment. Grade 5 titanium surfaces appear tougher, whereas anodized surfaces tend to lose the anodizing layer. (4) Conclusions: further studies will be needed to fully understand how these decontaminated surfaces affect the adhesion, proliferation and differentiation of fibroblasts and osteoblasts.
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Members of the family Meloidae are known to produce cantharidin, a highly toxic monoterpene found in their hemolymph and exuded as droplets capable of deterring many predators. As a nuptial gift, males transfer large amounts of cantharidin to females via a spermatophore, which is formed by specific accessory glands containing high concentrations of this terpene. Using light, electron and ion beam microscopy, the ultrastructural features of the three pairs of male accessory glands as well as the glandular part of the vasa deferentia were comparatively investigated in seven species of blister beetles belonging to five different tribes and two subfamilies. All gland pairs examined share common features such as mesodermal derivation, the presence of muscle sheath, a developed rough endoplasmic reticulum, abundant mitochondria, secretory vesicles, and microvillated apical membranes. Within the same species, glands exhibit distinctive features, suggesting that each pair is responsible for the formation of a specific substance. The vasa deferentia, while showing many similarities within the family, often exhibit features unique to each of the individual species investigated, whereas the accessory glands of the first and second pairs display the highest degree of ultrastructural variability. A comparison across the species shows an interesting constancy limited to ultrastructural features in the third pair of accessory glands. The similarities and differences among the species are discussed in the light of the available literature and in relation to the potential role that blister beetles' male accessory glands could play in the storage and management of cantharidin.
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The surface topography of dental implants plays an important role in cell-surface interaction promoting cell adhesion, proliferation and differentiation influencing osseointegration. A hydrophilic implant leads to the absorption of water molecules and subsequently promotes the adhesion of cells to the implant binding protein. Dried salts on the implant surfaces allow one to store the implant surfaces in a dry environment while preserving their hydrophilic characteristics. This process has been identified as "dry technology". The aim of the present study is to describe from a micrometric and nanometric point of view the characteristics of this new bioactivated surface obtained using salts dried on the surface. Topographic analysis, energy-dispersive X-ray spectroscopy, and contact angle characterization were performed on the samples of a sandblasted and dual acid-etched surface (ABT), a nanosurface (Nano) deriving from the former but with the adding of salts air dried and a nanosurface with salts dissolved with distilled water (Nano H2O). The analysis revealed promising results for nanostructured surfaces with increased wettability and a more articulated surface nanotopography than the traditional ABT surface. In conclusion, this study validates a new promising ultra-hydrophilic nano surface obtained by sandblasting, double acid etching and surface salt deposition using dry technology.
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Environmental thermal conditions play a major role at all levels of biological organization; however, there is little information on noxious high temperature sensation crucial in behavioral thermoregulation and survival of small ectothermic animals such as insects. So far, a capability to unambiguously encode heat has been demonstrated only for the sensory triad of the spike bursting thermo- and two bimodal hygro-thermoreceptor neurons located in the antennal dome-shaped sensilla (DSS) in a carabid beetle. We used extracellular single sensillum recording in the range of 20-45°C to demonstrate that a similar sensory triad in the elaterid Agriotes obscurus also produces high temperature-induced bursty spike trains. Several parameters of the bursts are temperature dependent, allowing the neurons in a certain order to encode different, but partly overlapping ranges of heat up to lethal levels in a graded manner. ISI in a burst is the most useful parameter out of six. Our findings consider spike bursting as a general, fundamental quality of the classical sensory triad of antennal thermo- and hygro-thermoreceptor neurons widespread in many insect groups, being a flexible and reliable mode of coding unfavorably high temperatures. The possible involvement of spike bursting in behavioral thermoregulation of the beetles is discussed. By contrast, the mean firing rate of the neurons in regular and bursty spike trains combined does not carry useful thermal information at the high end of noxious heat.