Preimplantation development of in vitro-produced bovine embryos treated with hydroxychloroquine.

Hydroxychloroquine (HCQ) is a safe antimalarial drug but its overdosage or inappropriate use, such as during the pandemic, may cause adverse effects once this drug is considered a potent inhibitor of autophagy. Information about HCQ's effects on the reproductive field, including gametes and initial embryos, is limited. In this study, we evaluated the effect of HCQ (1, 6, 12, and 24 µM) on pre-implantation embryo development, autophagy, and apoptosis of bovine embryos produced in vitro. A dose-response experiment showed a reduction (p < 0.05) in cleavage only at the highest concentration. Blastocyst rate was gradually reduced (p < 0.05) with the increase of HCQ dosage starting at 6 µM, with no embryo formation occurring at 24 µM. Further analysis showed that embryos treated with 12 µM of HCQ had a higher (p < 0.05) accumulation of acidic autophagic vesicles on Days 5 and 7 of development and a higher (p < 0.01) apoptotic index on Day 7. To our knowledge, this is the first study to evaluate the effects of HCQ on embryo pre-implantation development in mammals. The results contribute with more information related to the study of autophagy in embryology as well as add some discussion on HCQ toxicology and its effects on reproductive cells.

Lead and zinc concentrations in household dust and toenails of the residents (Estarreja, Portugal): a source-pathway-fate model.

This paper describes a methodology developed to assess and apportion probable indoor and outdoor sources of potentially toxic elements while identifying chemical signatures in the household dust collected from private homes in an industrial city (Estarreja, central Portugal). Oral bioaccessibility estimates and the chemical composition of toenail clippings were used to assess indoor dust ingestion as a potential exposure pathway and further investigate exposure-biomarker relationships. Indoor and paired outdoor dust samples were collected from each household. A total of 30 individuals, who provided toenail clippings and a self-reported questionnaire, were recruited for the study. Total concentrations of 34 elements, including lead and zinc, were determined in washed toenail samples and household dust via Inductively Coupled Plasma-Mass Spectrometry. The oral bioaccessibility was estimated using the Unified BARGE Method. The enrichment factor shows that lead was enriched (10 < EF < 100) while zinc (EF > 100) was anomalously enriched in the household dust, thus indicating potential exposure in the home environment. The results from principal component analysis coupled to cluster analysis and linear discriminant analysis suggested that mixed contamination derived from multiple sources with a predominance of biomass burning. Stepwise multiple linear regression analysis was performed to model toenail data using the indoor dust elemental composition. Whereas the model obtained for lead was not reliable, indoor dust zinc and antimony contents arose as good predictors of toenail zinc. The exposure-biomarker relationships seem to be influenced by the oral bioaccessibility of the elements.

Investigating relationships between biomarkers of exposure and environmental copper and manganese levels in house dusts from a Portuguese industrial city.

This study reports on data obtained from a pilot survey focusing on house dust and toenail metal(loids) concentrations in residents living in the industrial city of Estarreja. The study design hereby described aims at investigating relationships between human toenails and both copper and manganese levels in settled house dusts. A total of 21 households and 30 individuals were recruited for the pilot study: 19 households corresponding to 27 residents living near the industrial complex, forming the exposed group, plus 2 households and 3 residents from residential areas with no anticipated environmental contaminants that were used for comparison. Factorial analysis was used for source identification purposes. Investigation on the potential influence of environmental factors over copper and manganese levels in the toenails was carried out via questionnaire data and multiple correspondence analysis. The results show that copper concentrations are more elevated in the indoor dusts, while manganese concentrations are more elevated in the outdoor dust samples. The geometrical relationships in the datasets suggest that the backyard soil is a probable source of manganese to the indoor dust. Copper and manganese contents in the toenail clippings are more elevated in children than in adults, but the difference between the two age groups is not statistically significant (p > 0.05). Investigation of environmental factors influencing the exposure-biomarker association indicates a probable relationship between manganese contents in indoor dust and manganese levels in toenail clippings, a result that is partially supported by the bioaccessibility estimates. However, for copper, no relationship was found between indoor dusts and the biomarkers of exposure.

Geochemistry, mineralogy, solid-phase fractionation and oral bioaccessibility of lead in urban soils of Lisbon.

An urban survey of Lisbon, the largest city in Portugal, was carried out to investigate its environmental burden, emphasizing metallic elements and their public health impacts. This paper examines the geochemistry of lead (Pb) and its influence on human health data. A total of 51 soil samples were collected from urban recreational areas used by children to play outdoors. The semi-quantitative analysis of Pb was carried out by inductively coupled plasma mass spectrometry after an acid digestion. X-ray diffraction was used to characterize the soil mineralogy. The solid-phase distribution of Pb in the urban soils was investigated on a subset of 7 soils, out of a total of 51 samples, using a non-specific sequential extraction method coupled with chemometric analysis. Oral bioaccessibility measurements were obtained using the Unified BARGE Method developed by the Bioaccessibility Research Group of Europe. The objectives of the study are as follows: (1) investigation of Pb solid-phase distribution; (2) interpretation of Pb oral bioaccessibility measurements; (3) integration of metal geochemistry with human health data; and (4) understanding the influence of geochemistry and mineralogy on oral bioaccessibility. The results show that the bioaccessible fraction of Pb is lower when major metal fractions are associated with less soluble soil phases such as Fe oxyhydroxides, and more increased when the metal is in the highly soluble carbonate phase. However, there is some evidence that the proportion of carbonates in the soil environment is also a key control over the oral bioaccessibility of Pb, irrespective of its solid-phase fractionation.

Metal fractionation of cadmium, lead and arsenic of geogenic origin in topsoils from the Marrancos gold mineralisation, northern Portugal.

The Marrancos gold mineralisation has a chemical assemblage of Fe-As-Se-Bi-Au-Ag-Te-(Cu-Pb-Zn-Sn-W). The -200 mesh of 144 topsoil samples was analysed by ICP-MS to determine total contents of 53 elements that include potentially harmful elements like Cd, As and Pb. The soil geochemistry shows that some trace elements occur in considerably high concentrations. On the basis of data for total metal concentrations, 10 topsoil samples were selected to carry out a metal fractionation study using a selective extraction method. A set of four leaches of increasing strength was used sequentially in the soil samples. Across the study area, there is some evidence of past mining and exploration activity, indicating that these soils may be locally disturbed. The shallow mineralised quartz veins were exploited for gold by the Romans. Several galleries were constructed during the 2nd World War, probably for the exploration of quartz-cassiterite-wolframite veins. However, the main mineralised body in depth was never explored. The results of metal fractionation show different partitions for the three elements. Total Cd concentrations in these soils are low, with a median value of 0.1 mg/kg. In average, 12% of total Cd is adsorbed by clays and/or co-precipitated with carbonates, and 19% is bounded to Fe-Mn oxyhydroxides. However, the low concentrations indicate that the metal does not represent an immediate risk to human health. For Pb, metal fractionation shows that, on average, 22% of Pb is adsorbed by amorphous Fe and Mn oxides, but the samples from the northern part of the area have the major fractions of Pb in soluble forms. The low probability of exposure in this part of the study area decreases the risk posed by this heavy metal. Total As concentrations in the Marrancos soils are extremely high. A large area has As concentrations above 1,000 mg/kg. For As, metal fractions in the sulphide phase vary between 84 and 98% in the studied samples. But one sample has 20% of total As in easily reducible forms, corresponding to a partial concentration of 1,800 mg/kg that has a high probability of being bioavailable. The most labile As forms occur at the southern part of the area, where the probability of exposure is higher and the risk of human health increases in the same order. From the three studied potentially harmful elements, As is certainly the element of concern.

Lead availability in soils from Portugal's Centre Region with special reference to bioaccessibility.

Previous environmental biomonitoring studies indicated higher environmental lead (Pb) pollution levels at the districts of Aveiro and Leiria (Portugal). In evaluating the risk for human health, which is associated with contaminated soils after oral uptake, total soil concentrations have generally been held against criteria established from toxicological studies based upon the assumption that the uptake of the contaminant is similar in the toxicological studies and from the soils assessed. This assumption is not always valid, as most toxicological studies are carried out with soluble forms of the contaminants, whereas many soil contaminants are or become embedded in the soil matrix and thus exhibit limited availability. This study intends to estimate the soluble fraction of Pb in the soils from central Portugal, and to assess the bioaccessibility of Pb and, hence, infer exposure and risk for human health. Yet, as the physical-chemical properties of the soil exert some control over the solubility of Pb in the surface environment, the relation between such soil properties and the estimated soluble and/or bioaccessible fractions of Pb is also investigated. Other objective, with a more practical nature, was to give some contribution to find a suitable in vitro mimetic of the gastrointestinal tract environment. The results indicate relatively low total metal concentrations in the soils, even if differences between regions were observed. The Aveiro district has the higher total Pb concentration and the metal is in more soluble forms, that is, geoavailable. Soils with higher concentrations of soluble Pb show higher estimates of bioaccessible Pb. Soil pH seems to influence human bioaccessibility of Pb.

Microbial response to environmental gradients in a ceramic-based diffusion system.

A solid, porous matrix was used to establish steady-state concentration profiles upon which microbial responses to concentration gradients of nutrients or antimicrobial agents could be quantified. This technique relies on the development of spatially defined concentration gradients across a ceramic plate resulting from the diffusion of solutes through the porous ceramic matrix. A two-dimensional, finite-element numerical transport model was used to predict the establishment of concentration profiles, after which concentration profiles of conservative tracers were quantified fluorometrically and chemically at the solid-liquid interface to verify the simulated profiles. Microbial growth responses to nutrient, hypochloride, and antimicrobial concentration gradients were then quantified using epifluorescent or scanning confocal laser microscopy. The observed microbial response verified the establishment and maintenance of stable concentration gradients along the solid-liquid interface. These results indicate the ceramic diffusion system has potential for the isolation of heterogeneous microbial communities as well as for testing the efficacy of antimicrobial agents. In addition, the durability of the solid matrix allowed long-term investigations, making this approach preferable to conventional gel-stabilized systems that are impeded by erosion as well as expansion or shrinkage of the gel.

Spatial patterns of dispersion and pollution sources for arsenic at Lousal mine, Portugal.

This study uses geostatistical modelling to identify hot spots of metal contamination produced by hazardous mining tailings from an inoperative Portuguese mine. Lousal is one of the many massive sulphide deposits of the Iberian Pyrite Belt. The mine is now closed, but the heavy metal enriched tailings remain at the site. Applying geostatistics to the diagnosis of this potentially contaminated area we aim to assess the quality of the soil, namely through a risk probability mapping for arsenic, for a better knowledge about the vulnerability of the soil to arsenic contamination. To achieve this aim, the initial variable was transformed into two indicator variables using the risk-based standards (intervention values) for soils, as proposed by Swartjes (1999), as the limit value. To account for the spatial structure, sample variograms were computed for the main directions of the sampling grid and a spherical model was fitted to the variables (arsenic raw data and indicator variables). The parameters of the model were used in the estimation process. To assess the vulnerability of the soil towards the mining works, a soil quality probability mapping for arsenic was carried out showing contamination probabilities in the area. The use of indicator kriging, as an alternative to other more complex kriging methods, in the soil data of the Lousal mine, produced unbiased soil quality maps.

Application of geostatistical methods to arsenic data from soil samples of the Cova dos Mouros mine (Vila Verde-Portugal).

A total of 286 soil samples were collected in the Cova dos Mouros area. All samples were dry sieved into the <200 mesh size fraction and analysed for Fe, Cu, Zn, Pb, Co, Ni, Bi and Mn by atomic absorption spectrometry (AAS) and for As, Se, Sb and Te by atomic absorption spectrometry-hydrid generation (AAS-HG). Only the results of arsenic are discussed in this paper although the survey was extended to all analysed chemical elements. The purpose of this study was to make a risk probability mapping for arsenic that would allow better knowledge about the vulnerability of the soil to arsenic contamination. To achieve this purpose, the initial variable was transformed into an indicator variable using as thresholds the risk-based standards (intervention values) for soils, as proposed by [Swartjes 1999. Risk based assessment of soil and groundwater quality in the Netherlands: Standards and remediation. J. Geochem. Explor.73 1-10]. To account for spatial structure, sample variograms were computed for the main directions of the sampling grid and a spherical model was fitted to each sample variogram (arsenic variable and indicator variables). The parameters of the spherical model fitted to the arsenic variable were used to predict arsenic concentrations at unsampled locations. A risk probability mapping was also done to assess the vulnerability of the soil towards the mining works. The parameters of the spherical model fitted to each indicator variable were used to estimate probabilities of exceeding the corresponding threshold. The use of indicator kriging as an alternative to ordinary kriging for the soil data of Cova dos Mouros produced unbiased probability maps that allowed assessment of the quality of the soil.

Distribution of trace metals in moss biomonitors and assessment of contamination sources in Portugal.

A biomonitoring survey using the moss species [Hypnum cupressiforme Hedw. and Scelopodium touretii (Brid.) L. Kock] was performed in the whole territory of Portugal, in order to evaluate the atmospheric deposition of the following elements: Cd. Cr, Cu, Fe, Mn, Ni, Pb and Zn. The concentrations of the same elements were also obtained in two types of soil samples, collected under the moss and in nearby plots without any plant coverage, and relationships between moss and soil concentrations was investigated using the multivariate statistical method of Co-inertia Analysis. Also, relationships between concentrations in moss and several anthropogenic, geologic, pedologic and environmental parameters were screened using the same method of Co-inertia Analysis. Higher concentrations of Cu, Pb and Zn were found in areas of higher population density, with higher gasoline consumption, while higher values of Fe and Cr occur in the driest region, with lower plant coverage, indicating strong contamination by resuspended soil particles. Results also show good agreement between moss and soil contents, even for elements with high contribution of anthropogenic sources. The spatial pattern in Portugal of element contents in mosses were also detected and discussed in relation to local contamination sources.