The earthworm Lumbricus terrestris is an anecic species living in natural soils but it is also a sentinel in pollution monitoring. Specimens of L.terrestris were exposed for 48 h though the filter ...paper contact test at 1 mg/mL of the chemicals: Lamotrigine (LMG), Cocaine (COC), Fipronil (FIP) and the pesticide bis-4-nitrophenyl phosphate (BNPP). After that period, the activities of Acetylcholinesterase, Glutathione S-transferase, Carboxylesterase (CE) using different substrates, and lipid peroxidation levels were evaluated in the exposed whole tissue earthworms. The results revealed differences only in CE activity, with 4-nitrophenyl butyrate (4NPB) and 1-naphthyl butyrate (1NB) the most responsive substrates to COC. The kinetic parameters of CE were characterized, for the first time, in whole tissue of this species. The chemical analysis by LC-MS/MS, confirmed the exposure to the parent compounds, identified metabolites and evidenced biotransformation pathways in earthworms. Metabolic reactions included oxidation (LMG and FIP), hydrolysis (COC and FIP) as well as glycosylation (LMG, COC and FIP). A hitherto unknown metabolite of LMG due to the conjugation with phenylalanine glutamine was formed. The in vivo results on CE activity with the specific inhibitor, BNPP, were confirmed in vitro. Moreover, in the in vitro approach, the inclusion of other contaminants of environmental concern supports the potential of CE as biomarker. This study identifies the main metabolites formed by earthworms for further in vivo exposures under more realistic conditions and the potential use of CE measures as biomarker of emerging contaminants.
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•First detection of lamotrigine, cocaine and fipronil metabolites in earthworms.•New metabolite of lamotrigine from its conjugation with phenylalanine glutamine.•Carboxylesterases are involved in cocaine metabolism in earthworms.•Detection of the main cocaine human metabolites in earthworms.•Carboxylesterases are potential biomarkers of contaminated soils.
Abstract Riboflavin (RF) is an essential water-soluble vitamin with unique biological and physicochemical properties such as transporterspecific cell internalization, implication in redox reactions, ...fluorescence and photosensitizing. Due to these features RF attracted researchers in various fields from targeted drug delivery and tissue engineering to optoelectronics and biosensors. In this review we will give a brief reminder of RF chemistry, its optical, photosensitizing properties, RF transporter systems and its role in pathologies. We will point a special attention on the recent findings concerning RF applications in nanotechnologies such as RF functionalized nanoparticles, polymers, biomolecules, carbon nanotubes, hydrogels and implants for tissue engineering.
Transepithelial electrical resistance (TEER) measurements are regularly used in in vitro models to quantitatively evaluate the cell barrier function. Although it would be expected that TEER values ...obtained with the same cell type and experimental setup were comparable, values reported in the literature show a large dispersion for unclear reasons. This work highlights a possible error in a widely used formula to calculate the TEER, in which it may be erroneously assumed that the entire cell culture area contributes equally to the measurement. In this study, we have numerically calculated this error in some cell cultures previously reported. In particular, we evidence that some TEER measurements resulted in errors when measuring low TEERs, especially when using Transwell inserts 12 mm in diameter or microfluidic systems that have small chamber heights. To correct this error, we propose the use of a geometric correction factor (GCF) for calculating the TEER. In addition, we describe a simple method to determine the GCF of a particular measurement system, so that it can be applied retrospectively. We have also experimentally validated an interdigitated electrodes (IDE) configuration where the entire cell culture area contributes equally to the measurement, and it also implements minimal electrode coverage so that the cells can be visualized alongside TEER analysis.
Cephalopods are a group of marine invertebrates that have received little attention as sentinel species in comparison to other molluscs, such as bivalves. Consequently, their physiological and ...biochemical xenobiotic metabolism responses are poorly understood. Here we undertake a comparative analysis of the enzymatic activities involved in detoxification reactions and neural transmission in the digestive tract of two commercial cephalopods: the Common octopus, Octopus vulgaris, and the European cuttlefish, Sepia officinalis. For methodological purposes, several common B-esterases (five carboxylesterase (CE) substrates and three cholinesterase (ChE) determinations) were assayed as a proxy of metabolic and neuronal activities, respectively. Four components of the digestive tract in each species were considered: salivary glands, the stomach, the digestive gland and the caecum. The in vitro responses of digestive gland homogenates to model chemicals and contaminants of environmental concern were contrasted between both cephalopod species. The baseline biochemical activities in the four digestive tract components were also determined. Moreover, in order to validate the protocol, purified proteins, recombinant human CE (CE1 and CE2) and purified eel acetylcholinesterase (AChE) were included in the analysis. Overall, carboxylesterase activities were higher in octopus than in cuttlefish, with the activity quantified in the digestive tract components in the following order: digestive gland ≈ caecum > stomach ≈ salivary glands, with higher hydrolysis rates reached with naphthyl-derived substrates. In contrast, cuttlefish hydrolysis rates with ChE substrates were higher than in octopus. This trend was also reflected in a higher sensitivity to CE inhibitors in octopus and to AChE inhibitors in cuttlefish. Given the detoxification character of CEs and its protective role preventing AChE inhibition, octopus could be regarded as more efficiently protected than cuttlefish from neurotoxic exposures. A full characterisation of B-esterases in the digestive tract of the two common cephalopods is also provided.
•First time B-esterases characterisation in the digestive tract of two cephalopods.•Octopus carboxylesterase was sensitive to chemicals of environmental concern.•Cuttlefish acetylcholinesterase was sensitive to neurotoxic chemicals.•The flame retardant TBBPA interferes in vitro with carboxylesterases.•Cuttlefish AChE in vitro sensitive to organophosphorus flame retardants.
Recent studies, both in laboratory and sea conditions, have demonstrated damage after sound exposure in the cephalopod statocyst sensory epithelium, which secretes endolymph protein. Here, the ...proteomic analysis of the endolymph was performed before and after sound exposure to assess the effects of exposure to low intensity, low frequency sounds on the statocyst endolymph of the Mediterranean common cuttlefish (Sepia officinalis), determining changes in the protein composition of the statocyst endolymph immediately and 24 h after sound exposure. Significant differences in protein expression were observed, especially 24 h after exposure. A total of 37 spots were significantly different in exposed specimens, 17 of which were mostly related to stress and cytoskeletal structure. Among the stress proteins eight spots corresponding to eight hemocyanin isoforms were under-expressed possible due to lower oxygen consumption. In addition, cytoskeletal proteins such as tubulin alpha chain and intermediate filament protein were also down-regulated after exposure. Thus, endolymph analysis in the context of acoustic stress allowed us to establish the effects at the proteome level and identify the proteins that are particularly sensitive to this type of trauma.
AIM: The climate variability hypothesis (CVH) states that a positive relationship may exist between the breadth of thermal tolerance range and the level of climatic variability experienced by taxa ...with increasing latitude, especially in terrestrial ectotherms. Under CVH, we expected to find a correspondence between both thermal tolerance limits (CTmax and CTmin), ambient extreme temperature and the range sizes of species. We examined the validity of these predictions in a lowland tropical and a temperate tadpole assemblage. LOCATION: Lowland Neotropics (Bahia, Brazil) and Palaearctic (Iberian Peninsula and North Africa). METHOD: We employed phylogenetic eigenvector regression (PVR) and Pagel's lambda to analyse phylogenetic signals in CTmax and CTmin. We used phylogenetic regression analyses (PGLS) to test the relationships between thermal limits, range size and temperature predictors (measured at the macroscale and microhabitat levels) and phy‐ANOVA to compare both the physiological traits and thermal regimen in both tropical and temperate assemblages. RESULTS: We documented moderate‐to‐strong phylogenetic signal in both heat and cold tolerance. Temperate‐zone tadpoles had broader thermal tolerances than tropical ones. Thermal tolerance range was correlated with range sizes and was explained by seasonal thermal range predictors at the global scale. Both macro‐ and microclimate temperature variables provided the best predictive multivariate models of thermal limits at the global scale. Microclimatic predictors, however, were the main determinants of CTmax and CTmin variation at the local level of tropical and temperate communities respectively. MAIN CONCLUSIONS: Thermal tolerance range increases with latitude in tadpoles due to the higher increase in cold tolerance in temperate tadpoles. At the global scale, both macro‐ and microenvironment thermal information were reliable predictors of critical thermal limits and thermal tolerance range, as CVH predicts. However, thermal limits were best predicted by temperatures of the micro‐habitat at the regional level, thus suggesting that physiological thermal boundaries may be governed by thermal selection.
Copper recovery from printed circuit boards (PCB) from waste mobile phones was investigated using a two-step bioleaching process. The method consists of a first step where Fe(II) ions are ...biologically oxidised to Fe(III) by
Acidithiobacillus ferrooxidans.
Later, Fe (III) ions are put in contact with the PCBs for copper solubilisation. At the conditions tested in the present work, the Fe(II) bio-oxidation (first step) was almost completed in 48 h. Two different methods (filtration and sedimentation) for biomass separation before the second step were tested. No significance differences between both separation methods were observed in terms of the overall process efficiency. In both cases, using 7.5 g/L of e-waste concentration, copper recovery of 95–100% were obtained in only 48 h. In order to test an inexpensive and environmental friendly method to recovery the copper from the leachate solution, cementation of Cu (II) with metallic iron was performed. The copper powder obtained had purity of 64.8%.
Graphic Abstract
HIV viral reservoirs are established very early during infection. Resident memory T cells (T
) are present in tissues such as the lower female genital tract, but the contribution of this subset of ...cells to the pathogenesis and persistence of HIV remains unclear. Here, we show that cervical CD4
T
display a unique repertoire of clusters of differentiation, with enrichment of several molecules associated with HIV infection susceptibility, longevity and self-renewing capacities. These protein profiles are enriched in a fraction of CD4
T
expressing CD32. Cervical explant models show that CD4
T
preferentially support HIV infection and harbor more viral DNA and protein than non-T
. Importantly, cervical tissue from ART-suppressed HIV
women contain high levels of viral DNA and RNA, being the T
fraction the principal contributor. These results recognize the lower female genital tract as an HIV sanctuary and identify CD4
T
as primary targets of HIV infection and viral persistence. Thus, strategies towards an HIV cure will need to consider T
phenotypes, which are widely distributed in tissues.
A total of 164 blood samples from 16 clinically healthy bottlenose dolphins (Tursiops truncatus), were obtained from an aquarium in Spain between 2019 and 2020, as part of their preventive medicine ...protocol. In addition to conventional haematological and biochemical analyses, plasmatic B-esterase activities were characterised to determine the potential application of such analyses in wild counterparts. The hydrolysis rates for the substrates of acetylcholinesterase (AChE), butyrylcholinesterase (BuChE) and carboxylesterase (CE) activity in plasma were measured, the last using two commercial substrates, p-nitrophenyl acetate (pNPA) and p-nitrophenyl butyrate (pNPB). Activity rates (mean ± SEM in nmol/min/mL plasma) were (in descending order): AChE (125.6 ± 3.8), pNPB-CE (65.0 ± 2.2), pNPA-CE (49.7 ± 1.1) and BuChE (12.8 ± 1.3). These values for dolphins are reported in here for the first time in this species. Additionally, the in vitro sensitivity of two B-esterases (AChE and pNPB-CE) to chemicals of environmental concern was determined, and the protective role of plasmatic albumin assessed. Out of the B-esterases measured in plasma of dolphin, AChE activity was more responsive in vitro to pesticides, while CEs had a low response to plastic additives, likely due to the protective presence of albumin. However, the clear in vitro interaction of these environmental chemicals with purified AChE from electric eels and recombinant human hCEs (hCE1 and hCE2) and albumin, predicts their impact in other tissues that require in vivo validation. A relationship between esterase-like activities and health parameters in terrestrial mammals has already been established. Thus, B-esterase measures could be easily included in marine mammal health assessment protocols for dolphins as well, once the relationship between these measures and the animal's fitness has been established.
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•First time characterisation of plasmatic B-esterases in dolphins.•Potential of B-esterases as biomarker of neurotoxic exposures in marine mammals.•TBBPA interferes with carboxylesterases.•Organophosphorous flame retardants interfere with carboxylesterases.•Plasmatic albumin prevents carboxylesterase inhibition by TBBPA.
The diagnosis of gastrointestinal stromal tumors (GISTs) has important prognostic and therapeutic implications. The specific diagnosis of GIST has to be based on immunocytochemistry. This study aimed ...to prospectively compare in a crossover manner the accuracy of endoscopic ultrasound (EUS)-guided fine-needle aspiration (EUS-FNA) and EUS-guided trucut biopsy (EUS-TCB) in the specific diagnosis of gastric GISTs. We hypothesized that EUS-TCB is superior to EUS-FNA in this respect.
Forty patients with gastric subepithelial tumors suspected on the basis of EUS of being a GIST underwent both EUS-FNA and EUS-TCB. The sequence in which the techniques were employed was randomly assigned to avoid bias.
Forty tumors were sampled (mean number of passes: 2.1 +/- 0.9 with EUS-TNB and 1.9 +/- 0.8 with EUS-FNA; P = not significant, NS). Final diagnoses were: GIST (n = 27), carcinoma (n = 2), leiomyoma (n = 1), schwannoma (n = 1), and no diagnosis possible (n = 9). Device failure occurred in 6 patients with EUS-TCB. A cytohistological diagnosis of mesenchymal tumor (n = 29) and carcinoma (n = 2) was made in 70 % of cases by EUS-FNA and in 60 % of cases by EUS-TCB ( P = NS). Among the samples that were adequate, immunohistochemistry could be performed in 74 % of EUS-FNA samples and in 91 % of EUS-TCB samples ( P = 0.025). When inadequate samples were included, the overall diagnostic accuracy of EUS-FNA was 52 % and that of EUS-TCB was 55 % ( P = NS). There were no complications.
EUS-TCB is not superior to EUS-FNA in GISTs because of the high rate of technical failure of trucut. However, when an adequate sample is obtained with EUS-TCB, immunohistochemical phenotyping is almost always possible. EUS-TCB can be safely performed in this set of patients.
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