Epigenetic regulation of folate receptor-α (FOLR1) in human placenta of preterm newborns.

INTRODUCTION: Folates are essential nutrients for fetal development and pregnancy outcomes; they are transported to the fetus during gestation through specific folate transporters located in the placenta. In preterm newborns, we previously showed a lower placental mRNA expression of FOLR1 along with higher folate and lower vitamin B12 cord blood levels. Thereby we aimed to explore FOLR1 methylation in placentas of preterm newborns and hypothesized an increased FOLR1 methylation associated with cord blood folates and vitamin B12 concentrations. METHODS: FOLR1 methylation and mRNA were determined by methylation sensitive - high resolution melting (MS-HRM) and by real-time PCR respectively, in two placental sides of placental tissues: maternal (basal, BP) and fetal plates (chorionic, CP) of moderate preterm infants (32-36 gestational age) and term birth (37-41 gestational weeks). Folates and vitamin B12 were determined by electrochemiluminescence in umbilical cord blood samples from term and preterm newborns. RESULTS: We found that in preterm newborns, FOLR1 mRNA was lower in both plates of placenta compared with term newborns (p < 0,05) and was negatively associated with methylation of FOLR1 in CP. Preterm newborns presented higher folate and lower vitB12 concentrations in cord blood which correlated with increased placental FOLR1 methylation. DISCUSSION: In preterm newborns, placental FOLR1 expression is regulated by epigenetic mechanisms and presumably by maternal concentrations of folate and vitamin B12.

Tadpoles of the horned frog Ceratophrys ornata exhibit high sensitivity to chlorpyrifos for conventional ecotoxicological and novel bioacoustic variables.

Previous studies reported that some species of the family Ceratophryidae are able to produce sounds during premetamorphic tadpole stages. We have now determined the effects of the cholinesterase-inhibiting insecticide chlorpyrifos (CPF) on sounds emitted by tadpoles of Ceratophrys ornata. Tadpoles were exposed individually in order to evaluate the progression of effects. Effects on sound production were complemented with common ecotoxicological endpoints (mortality, behavior, abnormalities and growth inhibition). C. ornata was found to be more sensitive than other native (= 67%, 50%) and non-native species (= 75%, 100%) considering lethal and sublethal endpoints, respectively. Effects on sounds appear along with alterations in swimming, followed by the presence of mild, then severe abnormalities and finally death. Therefore, sound production may be a good biomarker since it anticipates other endpoints that are also affected by CPF. Ceratophrys ornata is a promising new model species in ecotoxicology.

Lethal and sublethal effects of the pirimicarb-based formulation Aficida® on Boana pulchella (Duméril and Bibron, 1841) tadpoles (Anura, Hylidae).

Acute lethal and sublethal toxicity of the pirimicarb-based commercial formulation Aficida® were evaluated on Boana pulchella tadpoles. Whereas mortality was used as end point for lethality, frequency of micronuclei and other nuclear abnormalities as well as alterations in the frequency of erythroblasts in circulating blood as biomarkers for genotoxicity and cytotoxicity, respectively. Swimming, growth, developmental and morphological abnormalities were also employed as sublethal end points. Results show that the species is within the 13th percentile of the distribution of acute sensitivity of species to pirimicarb for aquatic vertebrates. Results revealed values of 23.78 and 101.45mg/L pirimicarb as LC5096h for GS25 and GS36 tadpoles, respectively. The most evident effects were related with the swimming activity with NOEC and LOEC values within the 0.005-0.39mg/L pirimicarb concentration range. Aficida® induced DNA damage at the chromosomal level by increasing micronuclei frequency and other nuclear abnormalities, i.e., lobbed and notched nuclei and binucleated cells. Cellular cytotoxicity was found after Aficida® treatment. The presence of abdominal oedemas in exposed organisms and thus flotation response of organisms could be proposed as a new sensitive exposure parameter. The multiple end point assessment approach used allowed a complete understanding the multi level of effects occurring by exposure to pirimicarb, at least in B. pulchella.

Hot infusions and risk of colorectal cancer in Uruguay: a case-control study.

BACKGROUND/OBJECTIVES: The evidence of possible roles for the most common hot infusions intake (tea and coffee) in the risk of colorectal cancer (CRC) needs additional data. Regarding 'mate' intake (infusion of Ilex paraguariensis herb), a previous multi-site study reported lack of association for its highest intake on CRC risk. The present study was conducted to better understand the associations between the intake of this and other infusions and CRC risk. SUBJECTS/METHODS: Patients (611 CRC incident cases and 2394 controls, all belonging to public hospitals) were interviewed through a questionnaire, including socio-demographic, reproductive and lifestyle variables, and a food-frequency questionnaire of 64 items, analyzing tea, 'mate' and coffee intake (consumer status, daily intake, age at start and at quit). Odds ratios (ORs) and 95% confidence intervals (95% CIs) were calculated through unconditional logistic regression, adjusting for relevant potential confounders. RESULTS: Tea and coffee intake displayed significant and inverse associations with CRC risk, mainly among men (OR=0.54, 95% CI 0.38-0.76 for tea and OR=0.59, 95% CI 0.41-0.85 for coffee). Mate intake showed a significant inverse association among women (OR=0.50, 95% CI 0.33-0.77), with a marginal heterogeneity between sexes (P=0.07). Concerning age strata, tea intake displayed inverse associations in all ages, whereas 'mate' and coffee intake showed stronger inverse associations for age ⩾70, suggesting a gradient along time. CONCLUSIONS: We found evidence of different significant inverse associations for tea, 'mate' and coffee intake and CRC risk. To our knowledge, this is the first epidemiologic study reporting inverse results on 'mate' intake and CRC, which are explained by a stronger association among women.European Journal of Clinical Nutrition advance online publication, 23 August 2017; doi:10.1038/ejcn.2017.130.

Occurrence and fate of pesticides in the Argentine stretch of the Paraguay-Paraná basin.

The Argentine stretch of the del Plata basin crosses regions devoted to extensive and intensive agriculture mostly with chemical pest control. The utilization of pesticides in the region has increased 900% in the last two decades associated with the introduction of biotech crops and direct-seeding techniques. Our objective was to study the occurrence, concentration, and fate of pesticides in surface water and bottom sediments of the principal tributaries and main watercourse of the Paraguay-Paraná River. We sampled 22 sites in the distal positions of the main affluents and main watercourse of the Paraná and report here results from two monitoring campaigns (2010-2012). Surface water and bottom sediments were analyzed according to standardized methods by matrix-solid-phase dispersion and liquid-liquid extraction, respectively. Twenty-three pesticide compounds were analyzed by gas chromatography. The results from both campaigns indicated a generalized but variable distribution in the concentrations detected throughout the basin. The ranges of total measured pesticide concentrations in water and sediments were, respectively, 0.004-6.62 µg/l and 0.16-221.3 µg/kg dry weight. Endosulfans, cypermethrin, and chlorpyrifos were ubiquitous compounds in both environmental compartments and quantitatively the most relevant. All concentrations detected in water were over the recommended guidelines for the protection of aquatic biota. The partitioning indicated a higher affinity for the sediments. Agricultural activity is the source of pesticide-pollution loads, transported by tributaries that reach the main watercourse and alter the quality of the aquatic ecosystem.

Water quality of the main tributaries of the Paraná Basin: glyphosate and AMPA in surface water and bottom sediments.

The Paraná River, the sixth largest in the world, is the receptor of pollution loads from tributaries traversing urban and industrialized areas plus agricultural expanses, particularly so in the river's middle and lower reaches along the Argentine sector. In the present study, we analyzed and discussed the main water quality parameters, sediment compositions, and content of the herbicide glyphosate plus its metabolite aminomethylphosphonic acid (AMPA) in water and sediments. Samples were obtained from distal positions in the principal tributaries of the Paraná and the main watercourse during surveys conducted in 2011 and 2012 to monitor the basin. Only 15 % of the water samples contained detectable concentrations of glyphosate at an average concentration of 0.60 µg/L, while no detectable levels of AMPA were observed. The herbicide and metabolite were primarily present in sediments of the middle and lower stretch's tributaries, there occurring at a respective average of 37 and 17 % in samples. The mean detectable concentrations measured were 742 and 521 µg/kg at mean, maximum, and minimum glyphosate/AMPA ratios of 2.76, 7.80, and 0.06, respectively. The detection of both compounds was correlated with the presence of sulfides and copper in the sediment matrix.

Genotoxic Effects Induced by Cd(+2), Cr(+6), Cu(+2) in the Gill and Liver of Odontesthes bonariensis (Piscies, Atherinopsidae).

Genotoxic effects of Cd(+2), Cr(+6), and Cu(+2) on the gill and liver of the Argentinean Silverside (Odontesthes bonariensis) were studied using the comet assay and in relation with the metal tissue accumulation. Fish were exposed to three waterborne concentrations of each metal for 2 and 16 days. Genotoxicity was assessed by the single cell gel electrophoresis (comet assay). After 2 days, significant increase of the genetic damage index (GDI) was only observed in the gill of fish exposed to Cr(+6) and Cu(+2), and the LOECs were 2160 nM and 921.1 nM, respectively. The gill LOEC for Cd(+2) by 16 days was 9.4 nM. In the liver, LOECs were obtained only for Cd(+2) and Cr(+6) and were 9.4 and 2160 nM, respectively. The three metals were able to induce genotoxic effects at environmentally relevant concentrations and the gill was the most sensitive organ.

Abnormalities in amphibian populations inhabiting agroecosystems in Northeastern Buenos Aires Province, Argentina.

The occurrence of abnormalities in amphibians has been reported in many populations, and its increase could be related to environmental pollution and habitat degradation. We evaluated the type and prevalence of abnormalities in 5 amphibian populations from agroecosystems with different degrees of agricultural disturbance (cultivated and reference areas). We detected 9 types of abnormalities, of which the most frequent were those occurring in limbs. The observed prevalence of abnormality in assessed populations from cultivated and reference areas was as follows: Rhinella fernandezae (37.1 and 10.2%, respectively), Leptodactylus latrans adults (28.1 and 9.2%) and juveniles (32.9 and 15.3%), and Hypsiboas pulchellus (11.6 and 2.8%). Scinax granulatus populations did not show abnormalities. Pseudis minuta, which was only detected in the reference area, exhibited a prevalence of 13.3%. For R. fernandezae, L. latrans, and H. pulchellus, generalized linear mixed models showed that prevalence of abnormalities was significantly higher (p < 0.05) in cultivated than in reference areas. L. latrans juveniles were more vulnerable to abnormalities than adults (p < 0.05). The presence of abnormalities in some species inhabiting different agroecosystems suggests that environmental stress factors might be responsible for their occurrence. While we detected pesticides (endosulfan, cypermethrin, and chlorpyrifos) and lower dissolved oxygen levels in ponds of the cultivated area, no data are currently available on how other factors, such as injuries from predators and parasite infections, vary by land use. Further research will be necessary to evaluate possible causes of abnormalities detected in the present study mainly in the context of factor interactions.

Processed meat consumption and risk of cancer: a multisite case-control study in Uruguay.

BACKGROUND: The role of processed meat in the aetiology of several cancers was explored in detail. METHODS: In the time period 1996-2004, a multisite case-control study was conducted in Montevideo, Uruguay. The study included 6 060 participants (3 528 cases and 2 532 controls) corresponding to cancers of the oral cavity, pharynx, oesophagus, stomach, colon, rectum, larynx, lung, female breast, prostate, urinary bladder, and kidney (renal cell carcinoma only). RESULTS: The highest odds ratios (ORs) were positively associated with cancers of the colon, rectum, stomach, oesophagus, and lung. With the exception of renal cell carcinoma, the remaining cancer sites were significantly associated with elevated risks for processed meat consumption. Furthermore, mortadella, salami, hot dog, ham, and salted meat were strongly associated with risk of several cancer sites. CONCLUSION: It could be concluded that processed meat intake could be a powerful multiorgan carcinogen.

Insecticide toxicity to Hyalella curvispina in runoff and stream water within a soybean farm (Buenos Aires, Argentina).

Toxicity to the locally dominant amphipod Hyalella curvispina was assessed in a first-order stream running through a cultivated farm. Cypermethrin, chlorpyrifos, endosulfan and glyphosate were sprayed throughout the studied period. Toxicity was assayed under controlled laboratory conditions with runoff and stream water samples taken from the field under steady state and flood conditions. Ephemeral toxicity pulses were observed as a consequence of farm pesticide applications. After pesticide application, runoff water showed 100% mortality to H. curvispina for 1 month, but no mortality thereafter. Toxicity persistence was shortest in stream water, intermediate in stream sediments and longest in soil samples. Runoff had a more important toxicity effect than the exposure to direct aerial fumigation. The regional environmental features determining fast toxicity dissipation are discussed.

Effect of aquatic vegetation on the persistence of cypermethrin toxicity in water.

Soybean production in Argentina comprises 15 million ha. Cypermethrin is the main insecticide applied amounting 150 g of active ingredient per hectare, thus representing roughly 2.3 thousand tons yearly released to the environment. Toxicity pulses have been observed in small streams draining agricultural basins, most of them sustaining macrophyte growth. Cypermethrin concentrations and its toxicity to the amphipod Hyalella curvispina was compared following an addition to laboratory mesocosms with and without a vegetation cover of the floating macrophyte Lemna sp. Both concentrations and toxicity decreased faster in the treatments covered with Lemna. Fast adsorption of the hydrophobic pesticide to the roots and fronds of Lemna was suggested.

Dietary folate intake and the risk of 11 types of cancer: a case-control study in Uruguay.

BACKGROUND: There is limited, but inconclusive, epidemiological evidence that high folate intake decreases the risk of colorectal and esophageal cancers. For other cancer sites, the evidence is even less consistent or extensive. MATERIALS AND METHODS: We conducted a case-control study of dietary folate intake and risk of 11 cancer sites in Uruguay between 1996 and 2004, including 3539 cancer cases and 2032 hospital controls. Unconditional logistic regression was used to estimate odds ratios (ORs) and 95% confidence intervals (CIs) of cancer associated with folate intake. RESULTS: In the multivariable model, there was a significant decrease in the risk of cancers of the oral cavity and pharynx (OR = 0.49, 95% CI 0.24-0.98), esophagus (OR = 0.29, 95% CI 0.14-0.60), upper aerodigestive tract (OR = 0.41, 95% CI 0.26-0.65), colorectum (OR = 0.42, 95% CI 0.23-0.76) and kidney (OR = 0.35, 95% CI 0.13-0.93) for the highest versus the lowest quartile of dietary folate intake. CONCLUSIONS: Our results not only confirm earlier findings of decreased risk of colorectal and esophageal cancers with a high dietary folate intake but also suggest decreased risk of several other cancers. However, we cannot exclude the possibility that residual confounding, multiple comparisons or other forms of bias could explain these results.

Toxicity persistence following an experimental cypermethrin and chlorpyrifos application in Pampasic surface waters (Buenos Aires, Argentina).

The Pampa plain is intensively cultivated, the genetically modified glyphosate-resistant soybean being the main culture. A small first-order stream draining a cultured farm was studied. During the dry 2007 spring, the channel became a wetland with remnant intercalated shallow pools. Toxicity persistence of cypermethrin and chlorpyrifos to the amphipod Hyallela curvispina was assessed following a simulated plane application in the recently formed pools. In situ exposures and laboratory toxicity tests were performed. An ephemeral toxicity pulse lasting 4 h after the application and mortality cessation following that was observed.

P450-aromatase activity and expression in human testicular tissues with severe spermatogenic failure.

There is evidence that impaired spermatogenesis is associated with an imbalance in the oestradiol/testosterone ratio and with Leydig cell (LC) dysfunction. In testis, P450-aromatase, encoded by CYP19, is responsible for the conversion of testosterone to oestradiol. The aims of this study were to quantify CYP19 mRNA expression, aromatase activity and protein localization, and to measure the oestradiol to testosterone ratio in testicular tissues of men with spermatogenic impairment. Twenty-four men with complete Sertoli cell-only syndrome (SCOS), 14 with focal SCOS, 14 with maturation arrest (MA), 8 with mixed atrophy and 30 controls with normal spermatogenesis were subjected to testicular biopsy. All subjects underwent a physical examination, cytogenetic and serum hormonal studies. Testicular CYP19 mRNA was quantified using real time RT-PCR. Testicular aromatase activity was measured using the (3)H(2)0 assay and protein expression was evaluated using immunohistochemistry. In cases, serum testosterone and oestradiol were normal, but the testosterone/LH ratio was lower compared with controls (p < 0.05). Aromatase was localized in the Leydig, Sertoli and germ cells of all tissues, although stronger intensity was observed in LC. Aromatase mRNA and activity were not altered in cases and correlated positively with LC number (r = 0.516 and r = 0.369; p < 0.008). The intratesticular oestradiol/testosterone ratio was elevated (p = 0.005) in complete SCOS patients compared with controls. In conclusion, testicular aromatase seems to be normal in most subjects with impaired spermatogenesis. However, an altered intratesticular oestradiol/testosterone ratio in some patients with complete SCOS suggests that aromatase is increased, which might contribute to Leydig cell dysfunction.

Impact of glyphosate on the development, fertility and demography of Chrysoperla externa (Neuroptera: Chrysopidae): ecological approach.

Few ecotoxicological studies have used life table analysis to evaluate the toxicity of pesticides on beneficial organisms. This study is the first report of the effect of the herbicide glyphosate on a predator insect, Chrysoperla externa, using a demographic approach. This predator is associated to soybean pests and has a potential role as a biological control agent in the Neotropical Region. The objective of this work was to evaluate the side-effects of glyphosate on the development, fertility and demography of C. externa, treated orally by ingestion of glyphosate-dipped eggs of Sitotroga cerealella in laboratory conditions. The data were analyzed using the age-stage, two-sex life table. Development from third larval instar to pupae and adult longevity were shorter in glyphosate-treatment than in the control. Adult pre-reproductive period was longer in glyphosate-treatment than in the control. Fecundity and fertility were deeply reduced, as well, being fertility greater affected. A high important reduction was registered in all population parameters. Most eggs from glyphosate-treated cohort looked abnormal, smaller than control, dehydrated and became black 2d after oviposition. In addition, adults developed tumours in the abdomen region at 20d after emergence, being the effect more drastic in females than males. It is beyond the scope of our study to speculate on the effects of this herbicide on C. externa field populations. However, it seems likely that populations under continuous use of glyphosate would be exposed at greater detrimental effects in the long term.

Cadmium exposure during pregnancy reduces birth weight and increases maternal and foetal glucocorticoids.

Cadmium exposure induces low birth weight through unknown mechanisms. Since low birth weight is associated to foetal exposure to high glucocorticoids (GC) concentrations, we hypothesized that low birth weight induced by prenatal exposure to Cd(2+) is, at least in part, mediated by higher foetal exposure to GC, specifically corticosterone, the main active GC in rodents. Pregnant rats were exposed to different dose of CdCl(2) administered in drinking water during the whole pregnancy period. At term, corticosterone was measured by enzyme immunoassay in maternal and foetal blood and in placental tissues. Cadmium was determined in placentas, maternal tissues (liver and kidney) and foetuses by inductively coupled plasma-mass spectrometry (ICP-MS). Placental 11beta-hydroxysteroid dehydrogenase type 2 (11beta-HSD2) activity and expression were determined by a radiometric conversion assay and quantitative RT-PCR respectively. Results demonstrated that 50 ppm of Cd(2+), which was accumulated in different maternal tissues but not in the foetus, reduced pup birth weights and increased plasma corticosterone concentrations, both in mother and foetus. Placental 11beta-HSD2 activity and expression did not change by the treatment. We conclude that 50 ppm of Cd(2+) administered during pregnancy, increase foetal corticosterone concentrations due, probably, to alterations of the regulatory mechanisms of placental barrier to GC causing a mild but significant reduced birth weight.

Effects of the glyphosate active ingredient and a formulation on Lemna gibba L. at different exposure levels and assessment end-points.

The use of formulations of the herbicide glyphosate in transgenic crops of the Pampa's plains of Argentina has extensively increased, though there is scarce information of its impact on non-target vascular plants from agro-ecosystem related surface waters. The sensitivity of a local clone of the macrophyte Lemna gibba L. to glyphosate active principle and Roundup Max formulation was studied in standardized laboratory conditions. Phytotoxic effects, considering the aquatic route, at a concentration range of glyphosate between 0.5 and 80 mg L(-1) as active ingredient during 10 days of exposure were assessed on plant population growth, frond growth, shape and number, total chlorophyll content and colony architecture. Exposure to 1 mg L(-1) of glyphosate (an expected environmental concentration) affects all the studied assessment endpoints, except for population growth and chlorophyll content. Equivalent concentrations of this herbicide as the active ingredient or RoundupMax indicate higher phytotocity of the formulation. Exposed plants at concentrations of herbicide between 1 and 7.5 mg L(-1) exhibit after two days a recovery of the multiplication rate. Frond aggregation and longer stipe was detected between 1 and 15 mg L(-1) of glyphosate, determining more open colony architecture. At higher concentrations of the herbicide fronds break-up. Comparisons with literature data indicate a higher sensitivity of the L. gibba local clone with respect to L. minor and algal species, and also a similar response to the herbicide in field experiments with the same species.

Acute and chronic effects of Cr(VI) on Hypsiboas pulchellus embryos and tadpoles.

In the last few years there has been great concern about declines in the abundance of several species of amphibians around the world. Among amphibians, anurans have a biphasic life cycle, with aquatic tadpoles and generally terrestrial adults, and they have an extremely permeable skin, making them excellent indicators of the health of the environment. A number of different causes have been suggested for the global decline of anurans, the pollution of their habitat by chemical stressors being considered one of the major factors. Among chemical stressors, heavy metals are known for their high toxicity at very low concentrations. This study assessed short- (96 h, 'acute') and long-term (1272 h, 'chronic') exposure to Cr(VI) at lethal (3 to 90 mg 1(-1)) and sublethal concentrations (0.001 to 12 mg 1(-1)) on Hypsiboaspulchellus (previously called Hyla pulchella; see Faivovich et al. 2005) tadpoles (Fam. Hylidae) from central eastern Argentina. Fertilized eggs collected from a clean pond near La Plata (Buenos Aires Province) were used for acute and chronic toxicity testing. Assays were done under controlled laboratory conditions. Results of chronic exposure were used to assess the effect of factors such as toxicant concentration and age of organisms at the beginning of exposure on the response variables (growth, development and survival until metamorphosis). Results indicated a higher sensitivity to Cr(VI) of individuals first exposed as tadpoles than those first exposed as embryos during acute and chronic exposure. Exposure to the highest sublethal concentrations (6 to 12 mg 1(-1)) of the toxicant showed early inhibitory effects on growth of all treated organisms compensated at longer exposure periods with an increase in the growth rate compared to the control groups.