Experimental validation off-line a nonlinear controller for removal of chromium using non-living cells of Yarrowia lipolytica.

The removal of hexavalent chromium [Cr (VI)] using non-living cells of Yarrowia lipolytica was investigated. Batch and continuous studies on removal of Cr (VI) achieved 97% and 99% removal from aqueous phase, respectively. The specific uptake values at pH of 2 in batch process were 40.73 ± 1.3 mg/g and 30.09 ± 0.23 mg/g on non-living cells, when 100 and 200 mg/L of metal Cr (VI) concentrations were used. In order to investigate the regulation of Cr (VI) under continuous operation based on reaction volume numerically a new class of feedback controller from structure polynomial was designed. The proposed methodology was used to an experimentally kinetic model for a removal Cr (VI) from Yarrowia lipolytica biomass was showed satisfactory closed-loop performance the proposed controller. Starting from an off-line optimization performed in simulation, we present the controller implementation, focussing on the methodology required to could be suitable for implementation in real time. In our experimental results, we highlight some discrepancies between simulation and reality despite these differences, the controller managed to perform convergence to removal Cr (VI). Finally, the results validated with off-line samples suggest that the proposed control could be suitable for in application in potential scenarios for wastewater treatment.

A novel kinetic model for a cocoa waste fermentation to ethanol reaction and its experimental validation.

A non-segregated kinetic model is proposed to describe a fermentation process of agro-industrial residues derived via cocoa (mucilage juice) by Pichia kudriavzevii. The novel proposed hybrid model is based on a multiple coupling reaction mechanisms (structured) to describe the kinetics of substrate consumption, biomass, carbon dioxide, and ethanol, coupled to an unstructured model for the activity enzyme. The parameters of the kinetic model are estimated by non-linear least-squares curve fitting using the Marquardt-Levenberg algorithm. In addition, numerical simulations were compared with the experimental data via residual graphs. The effectiveness of the model was statistically evaluated using dimensionless efficiency coefficients under different initial conditions. A global sensitivity analysis was applied (Fisher's information matrix). The experimental results of the batch reactor showed a maximum ethanol concentration of 29 g/L, with a yield of 0.48 g-ethanol/g-glucose and a productivity of 0.30 g/L h. The method determined that the cell formation coefficient and the specific substrate consumption rate (θ1 and θ2) directly influence most of the states of our system. The proposed scheme is particularly suitable to assist in the rational design of cell factory properties or fermentation processes because it can represent the complex biochemistry in more detail and under different initial experimental conditions; the above reveals that the generated model is robust and can be considered for control and optimization purposes.

No evidence for effects of mill-grinding on δ13 C, δ15 N and δ34 S values in different marine taxa.

RATIONALE: The analysis of natural variation in light stable isotopes such as carbon (δ13 C), nitrogen (δ15 N) and sulfur (δ34 S) plays an important role in deepening our understanding of ecosystems. To avoid misinterpretation, robust results are required, where pre-treatment steps such as sample homogenization are crucial to guarantee representative samples. The grinding of samples using stainless steel balls in polypropylene tubes (e.g., laboratory bead-beater) has been identified as a potential source of bias. METHODS: We tested possible effects of mill-grinding (e.g., contamination) of samples of coastal marine taxa including primary producers, primary consumers and higher trophic level fish. We compared potential impacts of homogenization by mill-grinding with hand-grinding over an extended time on δ13 C, δ15 N and δ34 S values. RESULTS: One-way Welch's analysis of variance (ANOVA) showed that there were no statistical differences between methods for all the studied taxa. Also, repeated measures ANOVA showed no evidence of effects of grinding for extended times (from 30 to 120 seconds) for δ13 C, δ15 N and δ34 S values. CONCLUSIONS: We found no evidence that grinding samples in polypropylene tubes in a bead-beater resulted in any marked alteration of the isotopic composition on the studied samples, e.g., through contamination by plastic. As such, we consider mill-grinding as an appropriate method for the homogenization of samples from a range of different marine taxa, which under controlled conditions did not affect δ13 C, δ15 N and δ34 S analysis.

TGF-ß1/Smad2/3 signaling pathway modulates octreotide antisecretory and antiproliferative effects in pituitary somatotroph tumor cells.

Octreotide (OCT) is used to inhibit hormone secretion and growth in somatotroph tumors, although a significant percentage of patients are resistant. It has also been tested in nonfunctioning (NF) tumors but with poor results, with these outcomes having been associated with SSTR2 levels and impaired signaling. We investigated whether OCT inhibitory effects can be improved by TGF-ß1 in functioning and nonfunctioning somatotroph tumor cells. OCT effects on hormone secretion and proliferation were analyzed in the presence of TGF-ß1 in WT and SSTR2-overexpressing secreting GH3 and silent somatotroph tumor cells. The mechanism underlying these effects was assessed by studying SSTR and TGFßR signaling pathways mediators. In addition, we analyzed the effects of OCT/TGF-ß1 treatment on tumor growth and cell proliferation in vivo. The inhibitory effects of OCT on GH- and PRL-secretion and proliferation were improved in the presence of TGF-ß1, as well as by SSTR2 overexpression. The OCT/TGF-ß1 treatment induced downregulation of pERK1/2 and pAkt, upregulation of pSmad3, and inhibition of cyclin D1. In vivo experiments showed that OCT in the presence of TGF-ß1 blocked tumor volume growth, decreased cell proliferation, and increased tumor necrosis. These results indicate that SSTR2 levels and the stimulation of TGF-ß1/TGFßR/Smad2/3 pathway are important for strengthening the antiproliferative and antisecretory effects of OCT.

Estrogen receptor ß regulates the tumoral suppressor PTEN to modulate pituitary cell growth.

In this study, we focused on ERß regulation in the adenohypophysis under different estrogenic milieu, by analyzing whether ER modulates the phosphatase and tensin homolog deleted on chromosome 10 (PTEN) expression and its subcellular localization on anterior pituitary glands from Wistar rats and GH3 lactosomatotroph cells that over-expressed ERß. ERß was regulated in a cyclic manner, and underwent dynamic changes throughout the estrous cycle, with decreased ERß+ cells in estrus and under E2 treatment, but increased in ovariectomized rats. In addition, the ERα/ß ratio increased in estrus and under E2 stimulation, but decreased in ovariectomized rats. Double immunofluorescence revealed that lactotroph and somatotroph ERß+ were significantly decreased in estrus. Also, variations in the PTEN expression was observed, which was diminished with high E2 conditions but augmented with low E2 milieu. The subcellular localization of this phosphatase was cell cycle-dependent, with remarkable changes in the immunostaining pattern: nuclear in arrested pituitary cells but cytoplasmic in stimulated cells, and responding differently to ER agonists, with only DPN being able to increase PTEN expression and retaining it in the nucleus. Finally, ERß over-expression increased PTEN with a noticeable subcellular redistribution, and with a significant nuclear signal increase in correlation with an increase of cells in G0/G1 phase. These results showed that E2 is able to inhibit ERß expression and suggests that the tumoral suppressor PTEN might be one of the signaling proteins by which E2, through ERß, acts to modulate pituitary cell proliferation, thereby adapting endocrine populations in relation with hormonal necessities.

Secondary metabolites with antimicrobial activity produced by thermophilic bacteria from a high-altitude hydrothermal system.

Thermophilic microorganisms possess several adaptations to thrive in high temperature, which is reflected as biosynthesis of proteins and thermostable molecules, isolation and culture represent a great methodological challenge, therefore High throughput sequencing enables screening of the whole bacterial genome for functional potential, providing rapid and cost-effective information to guide targeted cultures for the identification and characterization of novel natural products. In this study, we isolated two thermophilic bacterial strains corresponding to Bacillus LB7 and Streptomyces LB8, from the microbial mats in the Atacama Desert. By combining genome mining, targeted cultures and biochemical characterization, we aimed to identify their capacity to synthesize bioactive compounds with antimicrobial properties. Additionally, we determined the capability to produce bioactive compounds under controlled in vitro assays and detected by determining their masses by Thin-Layer Chromatography/Mass Spectrometry (TLC/MS). Overall, both isolates can produce antimicrobial (e.g., Myxalamide C by-product) and antioxidants (e.g. Dihydroxymandelic Acid, Amide biotine and Flavone by-products) compounds. Bacillus LB7 strain possesses a more diverse repertoire with 51.95% of total metabolites unmatched, while Streptomyces LB8 favors mainly antioxidants, but has over 70% of unclassified compounds, highlighting the necessity to study and elucidate the structure of novel compounds. Based on these results, we postulate that the uncultured or rare cultured thermophiles inhabiting high-altitude hydrothermal ecosystems in the Atacama Desert offer a promising opportunity to the study of novel microbial bioactive compounds.

Chemical composition and antioxidant properties of native Ecuadorian fruits: Rubus glabratus Kunth, Vaccinium floribundum Kunth, and Opuntia soederstromiana.

The native berries of South America present promising marketing opportunities owing to their high antioxidant content, notably rich in anthocyanin and phenolic compounds. However, Ecuador's endemic fruits, primarily found in the wild, lack comprehensive data regarding their phytochemical composition and antioxidant capacity, underscoring the need for research in this area. Accordingly, this study evaluated the total phenolic, anthocyanin, flavonoid, resveratrol, ascorbic acid, citric acid, sugars, and antioxidant content of three native Ecuadorian fruits: mora de monte (Rubus glabratus Kunth), mortiño (Vaccinium floribundum Kunth), and tuna de monte (Opuntia soederstromiana). Determination of resveratrol, ascorbic acid, citric acid, and sugars was determined by HPLC analysis, and UPLC analysis was used to determine tentative metabolites with nutraceutical properties. Antioxidant capacity was assessed using cyclic voltammetry and the DPPH method; differential pulse voltammetry was used to evaluate antioxidant power. Analysis of results through UPLC-QTOF mass spectrometry indicated that R. glabratus Kunth and V. floribundum Kunth are important sources of various compounds with potential health-promoting functions in the body. The DPPH results showed the following antioxidant capacities for the three fruits: R. glabratus Kunth > O. soederstromiana > V. floribundum Kunth; this trend was consistent with the antioxidant capacity results determined using the electrochemical methods.

Environmentally relevant DEHP exposure during gestational and lactational period inhibits filamin a testicular expression.

Toxicological studies have revealed that DEHP exposure during pregnancy may induce developmental disorders, especially in male offspring, leading to morphological and functional alterations in the reproductive system by mechanisms that should be investigated. Thus, the aim of this work was to analyze the testicular toxicity induced by an environmentally relevant DEHP dose during development and its impact on FLNA, a protein that participates in the blood-testis barrier assembly. We used male Wistar rats exposed to DEHP during pregnancy and lactation. The results showed that DEHP exposure during development and lactation increased body weight, decreased gonadal weight and shortened anogenital distance. This phthalate induced morphological changes in the testis, suggestive of hypospermatogenesis. DEHP exposure decreased the number of FLNA positive cells and the expression of FLNA and claudin-1 in prepubertal testes. Furthermore, DEHP inhibited FLNA and claudin-1 protein expression in adult male rats. These results indicated that exposure to DEHP during gestation and lactation perturbed testis development and suggested that FLNA is a target protein of DEHP, possibly contributing to the phthalate-induced damage on BTB.

Antioxidant Capacity through Electrochemical Methods and Chemical Composition of Oenocarpus bataua and Gustavia macarenensis from the Ecuadorian Amazon.

This study evaluated the antioxidant properties and chemical composition of the seeds, pulp and peels of Ungurahua (Oenocarpus bataua) and Pasu (Gustavia macarenensis)-fruits, native to the Ecuadorian Amazon. The antioxidant capacity was measured by 1,1-diphenyl-2-picrylhydrazyl (DPPH) and cyclic voltammetry (antioxidant index 50 (AI50)) assays; differential pulse voltammetry was used to evaluate antioxidant power using the electrochemical index. The total phenolic content, as well as the yellow flavonoid and anthocyanin content, were quantified via spectrophotometry. In addition, the trans-resveratrol and ascorbic acid content were evaluated through high performance liquid chromatography (HPLC). Ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) was used to identify secondary metabolites with possible therapeutic properties. Results showed that the Pasu peel and seed extracts had the highest antioxidant capacity, followed by the Ungurahua peel; these results were consistent for both spectroscopic and electrochemical assays. HPLC and UPLC-MS analysis suggest that Oenocarpus bataua and Gustavia macarenensis are important sources of beneficial bioactive compounds.

Perinatal DEHP exposure modulates pituitary estrogen receptor α and ß expression altering lactotroph and somatotroph cell growth in prepuberal and adult male rats.

Phthalates are synthetic chemicals widely used to make polyvinylchloride (PVC) soft and flexible. Of these, Di-(2-ethylhexyl) phthalate (DEHP) is the most commonly used, with high human exposure occurring as early as the fetal developmental stage and affecting the endocrine system. We focused on the perinatal DEHP effects on pituitary estrogen receptor (ER) expression in male rats, explored their impact on lactotroph and somatotroph cell growth, and evaluated the direct effects of this phthalate on pituitary cell cultures. Our results showed that DEHP perinatal exposure was unable to modify the ERα+ pituitary cell number from prepuberal rats, but increased ERß+ cells. In adulthood, the pituitary ERα+ cells underwent a slight decrease with ERß showing the greatest changes, and with a significant increase observed in somatotroph cells. Also, in vitro, DEHP reduced the ERα+ cells, increased the percentage of ERß+ pituitary cells and modified the Ki67 index, as well as decreasing the lactotrophs and increasing the somatotroph cells. In conclusion, the present study showed that DEHP induced ER expression changes in normal pituitary glands from male rats in in vivo and in vitro conditions, suggesting that DEHP could differentially modulate lactotroph and somatotroph cell growth, possibly as a consequence of ER imbalance.

Total mercury bias in soil analysis by CV-AFS: causes, consequences and a simple solution based on sulfhydryl cotton fiber as a clean-up step.

This study examines the matrix effect over the trueness for determining total mercury (THg) using CV-AFS. We demonstrate that matrix interferences in soils and sediment samples cannot be eliminated by acid digestion and establish the use of sulfhydryl cotton fiber (SCF), a malleable, cheap and easy to synthesize fiber, as a mandatory solution capable to overcome this bias. Using the classic CV-AFS approach, an overestimation bias for THg recovery values of >140% in a certified reference material was reported. Interference metals test was conducted, thus discarding any influence of metals in the overestimation bias. Therefore, a clean-up step using SCF was proposed, and tests with synthesized fiber did not present a dispersion of >0.08 ng L-1. Moreover, validation was performed by analyzing three certified reference material and yielding mean recovery percentages of 100% ± 1%. A validated methodology was applied to ten environmental soil samples; THg values obtained varied from 129 to 384 ng g-1. Finally, a comparison between sample results obtained and reference method did not show any significant differences (p > 0.05), thus highlighting the efficacy of SCF-CV-AFS for THg quantification in environmental solid samples.

The phthalate DEHP modulates the estrogen receptors α and ß increasing lactotroph cell population in female pituitary glands.

Humans are exposed to numerous endocrine disruptors on a daily basis, which may interfere with endogenous estrogens, with Di-(2-ethylhexyl) phthalate (DEHP) being one of the most employed. The anterior pituitary gland is a target of 17ß-estradiol (E2) through the specific estrogen receptors (ERs) α and ß, whose expression levels fluctuate in the gland under different contexts, and the ERα/ß index is responsible for the final E2 effect. The aim of the present study was to evaluate in vivo and in vitro the DEHP effects on ERα and ß expression in the pituitary cell population, and also its impact on lactotroph and somatotroph cell growth. Our results revealed that perinatal exposure to DEHP altered the ERα and ß expression pattern in pituitary glands from prepubertal and adult female rats and increased the percentage of lactotroph cells in adulthood. In the in vitro system, DEHP down-regulated ERα and ß expression, and as a result increased the ERα/ß ratio and decreased the percentages of lactotrophs and somatotrophs expressing ERα and ß. In addition, DEHP increased the S + G2M phases, Ki67 index and cyclin D1 in vitro, leading to a rise in the lactotroph and somatotroph cell populations. These results showed that DEHP modified the pituitary ERα and ß expression in lactotrophs and somatotrophs from female rats and had an impact on the pituitary cell growth. These changes in ER expression may be a mechanism underlying DEHP exposure in the pituitary gland, leading to cell growth deregulation.

Mercury and methylmercury levels in soils associated with coal-fired power plants in central-northern Chile.

Mercury pollution is a worldwide problem, and is associated with a number of natural and anthropogenic processes. The present work, conducted in Chile, a country that has traditionally depended heavily on fossil fuels for power generation, examines total mercury (THg) and monomethylmercury (MMHg) concentrations in soils across different sites exposed to coal fired power plant emissions. Samples from four selected (Renca, Laguna Verde, Las Ventanas, Huasco) and 1 control (Quintay) sites were analyzed using cold vapour and fluorescence spectroscopy (CV-AFS) for THg determination and chromatographic separation with atomic fluorescence detection (DI-GC-AFS) was followed for speciation analysis. From the sites analyzed, Renca and Las Ventanas showed high concentrations of total mercury, exhibiting ranges between 135 - 568 and 94-464 ng g-1 respectively, while Laguna Verde and Huasco exhibited lower values ranged 5-27 and 9-44 ng g-1 respectively. Conversely, analysis of MMHg concentrations showed that only Renca site possessed high values, ranging between 0.1 and 3.0 ng g-1, resulting in this site being considered contaminated. Conversely, other sites showed minimal values comparable to the control site (0.024 ±â€¯0.003 ng g-1) in terms of MMHg concentrations. An analysis of the differences between MMHg and THg concentrations in contaminated sites, suggests an overall absence of methylation in soils of Las Ventanas, probably related to the very high levels of soil heavy metals, especially copper. Moreover, the influence of the composition and physicochemical properties of the different soils on the mobility of the species was assessed. Results obtained (as Log Kd) were 3.5 and 4.1 for Renca and Las Ventanas respectively, suggesting low mobility of mercury species in the environment for both sites. Finally, the data obtained allowed us to establish a first approximation of the differences in concentration and mobility of total and MMHg associated with coal fired power plants emission in central-northern Chile, an area previously understudied in a country heavily dependent on fossil-fuels.

Role of GPER in the anterior pituitary gland focusing on lactotroph function.

Ovarian steroids control a variety of physiological functions. They exert actions through classical nuclear steroid receptors, but rapid non-genomic actions through specific membrane steroid receptors have been also described. In this study, we demonstrate that the G-protein-coupled estrogen receptor (GPER) is expressed in the rat pituitary gland and, at a high level, in the lactotroph population. Our results revealed that ~40% of the anterior pituitary cells are GPER positive and ~35% of the lactotrophs are GPER positive. By immunocytochemical and immuno-electron-microscopy studies, we demonstrated that GPER is localized in the plasmatic membrane but is also associated to the endoplasmic reticulum in rat lactotrophs. Moreover, we found that local Gper expression is regulated negatively by 17ß-estradiol (E2) and progesterone (P4) and fluctuates during the estrus cycle, being minimal in proestrus. Interestingly, lack of ovarian steroids after an ovariectomy (OVX) significantly increased pituitary GPER expression specifically in the three morphologically different subtypes of lactotrophs. We found a rapid estradiol stimulatory effect on PRL secretion mediated by GPER, both in vitro and ex vivo, using a GPER agonist G1, and this effect was prevented by the GPER antagonist G36, demonstrating a novel role for this receptor. Then, the increased pituitary GPER expression after OVX could lead to alterations in the pituitary function as all three lactotroph subtypes are target of GPER ligand and could be involved in the PRL secretion mediated by GPER. Therefore, it should be taken into consideration in the response of the gland to an eventual hormone replacement therapy.

mPRs represent a novel target for PRL inhibition in experimental prolactinomas.

Membrane progesterone receptors are known to mediate rapid nongenomic progesterone effects in different cell types. Recent evidence revealed that mPRα is highly expressed in the rat pituitary, being primarily localized in lactotrophs, acting as an intermediary of P4-inhibitory actions on prolactin secretion. The role of mPRs in prolactinoma development remains unclear. We hypothesize that mPR agonists represent a novel tool for hyperprolactinemia treatment. To this end, pituitary expression of mPRs was studied in three animal models of prolactinoma. Expression of mPRs and nuclear receptor was significantly decreased in tumoral pituitaries compared to normal ones. However, the relative proportion of mPRα and mPRß was highly increased in prolactinomas. Interestingly, the selective mPR agonist (Org OD 02-0) significantly inhibited PRL release in both normal and tumoral pituitary explants, displaying a more pronounced effect in tumoral tissues. As P4 also regulates PRL secretion indirectly, by acting on dopaminergic neurons, we studied mPR involvement in this effect. We found that the hypothalamus has a high expression of mPRs. Interestingly, both P4 and OrgOD 02-0 increased dopamine release in hypothalamus explants. Moreover, in an in vivo treatment, that allows both, pituitary and hypothalamus actions, the mPR agonist strongly reduced the hyperprolactinemia in transgenic females carrying prolactinoma. Finally, we also found and interesting gender difference: males express higher levels of pituitary mPRα/ß, a sex that does not develop prolactinoma in these mice models. Taken together, these findings suggest mPRs activation could represent a novel tool for hyperprolactinemic patients, especially those that present resistance to dopaminergic drugs.

Oxidative stress and mitochondrial adaptive shift during pituitary tumoral growth.

The cellular transformation of normal functional cells to neoplastic ones implies alterations in the cellular metabolism and mitochondrial function in order to provide the bioenergetics and growth requirements for tumour growth progression. Currently, the mitochondrial physiology and dynamic shift during pituitary tumour development are not well understood. Pituitary tumours present endocrine neoplastic benign growth which, in previous reports, we had shown that in addition to increased proliferation, these tumours were also characterized by cellular senescence signs with no indication of apoptosis. Here, we show clear evidence of oxidative stress in pituitary cells, accompanied by bigger and round mitochondria during tumour development, associated with augmented biogenesis and an increased fusion process. An activation of the Nrf2 stress response pathway together with the attenuation of the oxidative damage signs occurring during tumour development were also observed which will probably provide survival advantages to the pituitary cells. These neoplasms also presented a progressive increase in lactate production, suggesting a metabolic shift towards glycolysis metabolism. These findings might imply an oxidative stress state that could impact on the pathogenesis of pituitary tumours. These data may also reflect that pituitary cells can modulate their metabolism to adapt to different energy requirements and signalling events in a pathophysiological situation to obtain protection from damage and enhance their survival chances. Thus, we suggest that mitochondria function, oxidative stress or damage might play a critical role in pituitary tumour progression.

Experimental and kinetic study for lead removal via photosynthetic consortia using genetic algorithms to parameter estimation.

This work presents an experimental-theoretical strategy for a batch process for lead removal by photosynthetic consortium, conformed by algae and bacteria. Photosynthetic consortium, isolated from a treatment plant wastewater of Tecamac (Mexico), was used as inoculum in bubble column photobioreactors. The consortium was used to evaluate the kinetics of lead removal at different initial concentrations of metal (15, 30, 40, 50, and 60 mgL-1), carried out in batch culture with a hydraulic residence time of 14 days using Bold's Basal mineral medium. The photobioreactor was operated under the following conditions: aeration of 0.5 vvm, 80 µmol m-2 s-1 of photon flux density and a photoperiod light/dark 12:12. After determining the best growth kinetics of biomass and metal removal, they were tested under different ratios (30 and 60%) of wastewater-culture medium. Additionally, the biomass growth (X), nitrogen consumption (N), chemical oxygen demand (COD), and metal removal (Pb) were quantified. Achieved lead removal was 97.4% when the initial lead concentration was up to 50 mgL-1 using 60% of wastewater. Additionally, an unstructured-type mathematical model was developed to simulate COD, X, N, and lead removal. Furthermore, a comparison between the Levenberg-Marquardt (L-M) optimization approach and Genetic Algorithms (GA) was carried out for parameter estimation. Also, it was concluded that GA has a slightly better performance and possesses better convergence and computational time than L-M. Hence, the proposed method might be applied for parameter estimation of biological models and be used for the monitoring and control process.

Critical evaluation of distillation procedure for the determination of methylmercury in soil samples.

In the present work, the efficiency of distillation process for extracting monomethylmercury (MMHg) from soil samples was studied and optimized using an experimental design methodology. The influence of soil composition on MMHg extraction was evaluated by testing of four soil samples with different geochemical characteristics. Optimization suggested that the acid concentration and the duration of the distillation process were most significant and the most favorable conditions, established as a compromise for the studied soils, were determined to be a 70 min distillation using an 0.2 M acid. Corresponding limits of detection (LOD) and quantification (LOQ) were 0.21 and 0.7 pg absolute, respectively. The optimized methodology was applied with satisfactory results to soil samples and was compared to a reference methodology based on isotopic dilution analysis followed by gas chromatography-inductively coupled plasma mass spectrometry (IDA-GC-ICP-MS). Using the optimized conditions, recoveries ranged from 82 to 98%, which is an increase of 9-34% relative to the previously used standard operating procedure. Finally, the validated methodology was applied to quantify MMHg in soils collected from different sites impacted by coal fired power plants in the north-central zone of Chile, measuring MMHg concentrations ranging from 0.091 to 2.8 ng g-1. These data are to the best of our knowledge the first MMHg measurements reported for Chile.

Inhibitory role of ERß on anterior pituitary cell proliferation by controlling the expression of proteins related to cell cycle progression.

Considering that the role of ERß in the growth of pituitary cells is not well known, the aim of this work was to determine the expression of ERß in normal and tumoral cells and to investigate its implications in the proliferative control of this endocrine gland, by analyzing the participation of cyclin D1, Cdk4 and p21. Our results showed that the expression of ERß decreased during pituitary tumoral development induced by chronic E2 stimulation. The 20 ± 1.6% of normal adenohypophyseal cells expressed ERß, with this protein being reduced in the hyperplastic/adenomatous pituitary: at 20 days the ERß+ population was 10.7 ± 2.2%, while after 40 and 60 days of treatment an almost complete loss in the ERß expression was observed (40 d: 1 ± 0.6%; 60 d: 2 ± 0.6%). The ERα/ß ratio increased starting from tumors at 40 days, mainly due to the loss of ERß expression. The cell proliferation was analyzed in normal and hyperplastic pituitary and also in GH3ß- and GH3ß+ which contained different levels of ERß expression, and therefore different ERα/ß ratios. The over-expression of ERß inhibited the GH3 cell proliferation and expression of cyclin D1 and ERα. Also, the ERß activation by its agonist DPN changed the subcellular localization of p21, inducing an increase in the p21 nuclear expression, where it acts as a tumoral suppressor. These results show that ERß exerts an inhibitory role on pituitary cell proliferation, and that this effect may be partially due to the modulation of some key regulators of the cell cycle, such as cyclin D1 and p21. These data contribute significantly to the understanding of the ER effects in the proliferative control of pituitary gland, specifically related to the ERß function in the E2 actions on this endocrine gland.

Fear-potentiated behaviour is modulated by central amygdala angiotensin II AT1 receptors stimulation.

Central nucleus of the amygdala (CeA) is one of the most important regulatory centres for the emotional processes. Among the different neurotransmitter systems present in this nucleus, AT1 receptors have been also found, but their role in the generation and modulation of emotions is not fully understood. The present work evaluated the effect of intra-amygdalar injection of losartan (AT1 receptor antagonist) and angiotensin II (Ang II) in the anxiety state induced by fear-potentiated plus maze in male Wistar rats. Fear in the elevated plus maze can be potentiated by prior inescapable footshock stress. The decrease in the time spent in the open arms induced by the inescapable footshock was totally prevented by losartan (4 pmol) administration in CeA. It was also found that Ang II (48 fmol) administration decreased the time spent in the open arms in animals with or without previous footshock exposure. The locomotor activity and grooming behaviour were also evaluated. The results obtained from the different parameters analyzed allowed us to conclude that the Ang II AT1 receptors in CeA are involved in the anxiety state induced by stress in the fear-potentiated plus-maze behaviour.