The regulation of the expression of prokineticin 1 and its receptors and its mechanism of action in the porcine corpus luteum.

Prokineticin 1 (PROK1) is an important factor in pregnancy establishment in pigs, acting at the embryo-maternal interface and the corpus luteum (CL). Estradiol-17ß (E2) is the primary pregnancy recognition signal in pigs, and its effects are augmented by luteotropic prostaglandin E2 (PGE2). On the contrary, prostaglandin F2α (PGF2α) exerts mainly a luteolytic effect. The present study aimed to elucidate whether E2, PGE2, and PGF2α regulate the expression of PROK1 and its receptors in the porcine CL and to determine the PROK1 effect on luteal endothelial cells and pathways that may be involved in this regulation. The effects of E2, PGE2, and PGF2α on the expressions of PROK1 and its receptors in the CL were studied using an in vitro model of ultrathin luteal tissue explants model. Additionally, the effects of E2 and PGE2 on the PROK1 system were determined using an in vivo approach, in which the hormones were administered into the uterine lumen to imitate their secretion by embryos. Endothelial cell proliferation was measured using the colorimetric method. E2 acting via estrogen receptors simulated the mRNA and protein expressions of PROK1 and PROKR1 in CL explants in vitro (p < 0.05). The simultaneous action of E2 with PGE2 enhanced the expression of luteal PROK1 mRNA in vitro (p < 0.05). Estradiol-17ß acting alone significantly increased PROK1 mRNA levels in vivo, whereas E2 simultaneously administered with PGE2 significantly elevated the PROK1 mRNA expression and PROKR1 mRNA and protein contents in CLs adjacent to uterine horns receiving hormonal infusion compared with CLs adjacent to placebo-treated uterine horns (p < 0.01). The PROK1 protein expression was significantly higher in the CLs of pigs treated with E2, PGE2, and E2 together with PGE2 than in the control group. PGF2α increased the PROK1 mRNA content in CLs on days 12 and 14 of the estrous cycle (p < 0.05). The expression of PROKR2 at the mRNA and protein levels remained unchanged in response to in vitro and in vivo treatments. PROK1 stimulated the proliferation of luteal endothelial cells by activating the MAPK, AKT, and mTOR pathways (p < 0.05). In summary, the luteal expressions of PROK1 and PROKR1 in early pregnancy are regulated by E2 and PGE2. PROK1 stimulates luteal angiogenesis by activating the MAPK, AKT, and mTOR pathways. The regulation of luteal PROK1 expression by PGF2α indicates PROK1's putative role during luteolysis. We conclude that PROK1-PROKR1 signaling supports luteal function during CL rescue in pregnancy in pigs.

Selected Fungicides as Potential EDC Estrogenic Micropollutants in the Environment.

An increasing level of pesticide exposition is being observed as a result of the consumption of large amounts of fruits, vegetables and grain products, which are key components of the vegetarian diet. Fungicides have been classified as endocrine-disrupting compounds, but their mechanisms of action have not yet been clarified. The effect of boscalid (B), cyprodinil (C) and iprodione (I) combined with Tamoxifen (T) and 17ß-estradiol (E2) on cell viability, cell proliferation, reporter gene expression, ROS content, the cell membrane's function, cell morphology and antioxidant enzymes gene expression in MCF-7 and T47D-KBluc cell lines were investigated. The cell lines were chosen due to their response to 17ß -estradiol. The selected fungicides are commonly used in Poland to protect crops against fungi. Our results revealed that the studied fungicides caused significant increases in cell viability and proliferation, and estrogenic activity was present in all studied compounds depending on their concentrations. Oxidative stress activated uncontrolled cancer cell proliferation by inducing ROS production and by inhibiting antioxidant defense. Our findings verify that the studied fungicides could possibly exhibit endocrine-disrupting properties and exposure should be avoided.

Impact of technological processes on tebuconazole reduction in selected cereal species and the primary cereal product, and dietary exposure assessment.

Contamination of cereals with tebuconazole (TEB) can affect the dietary risk assessment. This study investigates, for the first time, how mechanical, thermal, physical-chemical, and biochemical processes affect the TEB level in wheat, rye, and barley. The biochemical process of malting was the most effective for tebuconazole reduction (by 86%) in cereals. Thermal processes were also effective, i.e., boiling (70%) and baking (55%). These processes considerably decreased the concentration of tebuconazole, and Procesing Factors (PFs) were from 0.10 to 0.18 (malting), 0.56 to 0.89 (boiling), and 0.44 to 0.45 (baking), respectively. The concentration of TEB was not reduced after the application of mechanical processing. The risk was estimated in dietary exposure assessment on the basis of the highest reported levels of tebuconazole residues bread. At a high level of rye bread consumption, the potential exposure to tebuconazole reached only 3.5% and 2.7% in children and adults, respectively.

Removal of triazole and pyrethroid pesticides from wheat grain by water treatment and ultrasound-supported processes.

A simple way to reduce pesticides in cereal grains is to use washing methods. The challenge of this study was to evaluate the effectiveness of reduction of 3 triazole fungicides (difenoconazole, tebuconazole, tetraconazole) and 3 pyrethroid insecticides (beta-cyfluthrin, cypermethrin, deltamethrin) commonly used in wheat protection. Four different pre-washing methods (hot and cold water washing, twice water, and ultrasound-supported washing) were evaluated. The processing factor (PF) was calculated based on the concentration of pesticides determined by LC-MS/MS in the samples of cereal grains before and after the washing process. PFs were within the range 0.01-0.97. Time, teperature and ultrasound were factors influencing the efficiency of water treatment. The study showed that ultrasound-supported washing eliminated pesticide residues to a greater extent than ordinary washing. This process significantly affected or completely reduced concentrations of triazoles in wheat grains. The highest reduction of residues (99%) was received for tebuconazole and ultrasound washing with heating temperature of 60 °C for a total of 10 min. In all washing processes, pyrethroids were removed with lower efficiency than triazoles. The lowest residue reduction was obtained for cypermethrin and washing under cold water for 5 min (3%; PF = 0.97). Beta-Cyfluthrin showed only a 6-27% reduction regardless of the process (PF: 0.73-0.95). Using static analysis, the relationship between the properties of pesticides and the reduction of their concentration in cereals was clarified and showed a strong correlation.

Suitableness of SLM Manufactured Turbine Blade for Aerodynamical Tests.

This paper describes some insights on applicability of a Selective Laser Melting and Direct Metal Laser Sintering technology-manufactured turbine blade models for aerodynamic tests in a wind tunnel. The principal idea behind this research was to assess the possibilities of using 'raw' DLMS printed turbine blade models for gas-flow experiments. The actual blade, manufactured using the DLMS technology, is assessed in terms of surface quality (roughness), geometrical shape and size (outline), quality of counterbores and quality of small diameter holes. The results are evaluated for the experimental aerodynamics standpoint. This field of application imposes requirements that have not yet been described in the literature. The experimental outcomes prove the surface quality does not suffice to conduct quantitative experiments. The holes that are necessary for pressure measurements in wind tunnel experiments cannot be reduced below 1 mm in diameter. The dimensional discrepancies are on the level beyond acceptable. Additionally, the problem of 'reversed tolerance', with the material building up and distorting the design, is visible in elements printed with the DLMS technology. The results indicate the necessity of post-machining of the printed elements prior their experimental usage, as their features in the 'as fabricated' state significantly disturb the flow conditions.

Prokineticin 1 is a novel factor regulating porcine corpus luteum function.

Prokineticin 1 (PROK1) is a pleiotropic factor secreted by endocrine glands; however, its role has not been studied in the corpus luteum (CL) during pregnancy in any species. The present study aimed to investigate the contribution of PROK1 in regulating processes related to porcine CL function and regression: steroidogenesis, luteal cell apoptosis and viability, and angiogenesis. The luteal expression of PROK1 was greater on Days 12 and 14 of pregnancy compared to Day 9. PROK1 protein expression during pregnancy increased gradually and peaked on Day 14, when it was also significantly higher than that on Day 14 of the estrous cycle. Prokineticin receptor 1 (PROKR1) mRNA abundance increased on Days 12 and 14 of pregnancy, whereas PROKR2 elevated on Day 14 of the estrous cycle. PROK1, acting via PROKR1, stimulated the expression of genes involved in progesterone synthesis, as well as progesterone secretion by luteal tissue. PROK1-PROKR1 signaling reduced apoptosis and increased the viability of luteal cells. PROK1 acting through PROKR1 stimulated angiogenesis by increasing capillary-like structure formation by luteal endothelial cells and elevating angiogenin gene expression and VEGFA secretion by luteal tissue. Our results indicate that PROK1 regulates processes vital for maintaining luteal function during early pregnancy and the mid-luteal phase.

Novel insights into conceptus-maternal signaling during pregnancy establishment in pigs.

Pregnancy establishment in mammals, including pigs, requires coordinated communication between developing conceptuses (embryos with associated membranes) and the maternal organism. Porcine conceptuses signalize their presence by secreting multiple factors, of which estradiol-17ß (E2) is considered the major embryonic signal initiating the maternal recognition of pregnancy. During this time, a limited supply of prostaglandin (PGF2α) to the corpora lutea and an increased secretion of luteoprotective factors (e.g., E2 and prostaglandin E2 [PGE2]) lead to the corpus luteum's maintained function of secreting progesterone, which in turn primes the uterus for implantation. Further, embryo implantation is related to establishing an appropriate proinflammatory environment coordinated by the secretion of proinflammatory mediators including cytokines, growth factors, and lipid mediators of both endometrial and conceptus origin. The novel, dual role of PGF2α has been underlined. Recent studies involving high-throughput technologies and sophisticated experimental models identified a number of novel factors and revealed complex relationships between these factors and those already established. Hence, it seems that early pregnancy should be regarded as a sequence of processes orchestrated by pleiotropic factors that are involved in redundancy and compensatory mechanisms that preserve the essential functions critical for implantation and placenta formation. Therefore, establishing the hierarchy between all molecules present at the embryo-maternal interface is now even more challenging.

High and low temperature processing: Effective tool reducing pesticides in/on apple used in a risk assessment of dietary intake protocol.

Apples play an important role in everyone's diet and may contain pesticide residues that can pose a significant health problem for consumers. Various technological processes are promising methods for minimizing pesticide concentrations in fruit. Therefore, the subject of this comprehensive study was to investigate the effects of high-temperature (baking) and low-temperature (freeze-drying) processes on the change in the levels of nine fungicides in apples with skin and peeled. The investigated compounds belong to the chemical groups of benzimidazole (thiophanate methyl and carbendazim), phtalimide (captan and their metabolite tetrahydrophtalimid (THPI)), strobilurin (pyraclostrobin, trifloxystrobin) and triazole (difenoconazole, tebuconazole, tetraconazole). Processing factors (PF) were calculated for each pesticide-process-product combination. The results show that baking and freeze-drying generally reduced pesticide concentrations, with PFs ranging from 0.31 to 0.81 and 0.26 to 0.68, respectively. Apart from freeze-drying for carbendazim and baking for captan, PFs were above 1. Only for thiophanate-methyl, a complete reduction was observed, which resulted from complete degradation to carbendazim. The study also aimed to assess human risk according to the new strategy for different sub-populations with conversion using the 36 PFs obtained. The highest acute exposure (expressed as %ARfD) was obtained for tebuconazole in raw apples (initial concentration of 1.42 mg/kg; 400% ARfD) for Dutch toddlers. After food processing, this decreased to 284% (0.74 mg/kg, baking) and to 137% (0.37 mg/kg, freeze-drying), but was still above the safety limit. Similarly, for adults and the general French population for tebuconazole, the %ARfD was high as it reached the values of 104% (initial concentration of 0.89 mg/kg) in unprocessed apples, 73.9% after baking (0.73 mg/kg) and 35.6% after freeze-drying (0.35 mg/kg). The results indicate that food processing techniques can potentially be used to minimize the hazardous effects of pesticide residues on human health.

Novel role for conceptus signals in mRNA expression regulation by DNA methylation in porcine endometrium during early pregnancy†.

During early pregnancy, porcine conceptuses (the embryos with associated membranes) secrete estradiol-17ß (E2)-their major signal for maternal recognition of pregnancy-and prostaglandin E2 (PGE2). Both hormones induce prominent changes of the endometrial transcriptome in vivo. Studies on endometrial pathologies have shown that E2 affects gene expression by epigenetic mechanisms related to DNA methylation. Herein, we determined the effects of E2 and PGE2 alone, and a combined E2 + PGE2 treatment administered into the uterine lumen in vivo on the expression and activity of DNA-methyltransferases (DNMTs) and on CpG methylation patterns of selected genes in porcine endometrium. To compare the effect of treatment with the physiological effect of pregnancy, endometria from day 12 pregnant/cyclic gilts were included. Both E2 and PGE2 significantly reduced the expression of DNMTs. Likewise, the expressions of DNMT1 and DNMT3A were decreased on day 12 of pregnancy compared to the estrous cycle. DNMT activity increased in endometrial samples following E2 treatment and in gilts on day 12 of pregnancy. Treatment with E2 alone and/or simultaneously with PGE2 altered endometrial DNA methylation of CpG sites of ADAMTS20, ADH1C, BGN, PSAT1, and WNT5A. Different CpG methylation patterns of ADAMTS20, BGN, DMBT1, RASSF1, and WNT5A were found in the endometrium on day 12 of pregnancy compared to day 12 of the estrous cycle. Significant correlations were detected between CpG methylation and gene expression for ADAMTS20, ADH1C, BGN, DMBT1, PSAT1, and WNT5A. Our results indicate that CpG methylation induced by embryonic signals may contribute to regulating endometrial gene expression during pregnancy establishment.

Evaluation of titanium and silicon role in mitigation of fungicides toxicity in wheat expressed at the level of biochemical and antioxidant profile.

Biostimulators are compounds that are gaining attention due to their potential to support plant growth, however, less in known about their impact on plant biochemistry. Therefore, the main goal of this comprehensive study was to evaluate the effect of two inorganic biostimulators (titanium, Ti and silicon, Si) and five fungicides (propiconazole, cyproconazole, spiroxamine, tebuconazole, triadimenol) on biochemical and antioxidant status as well as mycotoxin profile in wheat (Triticum aestivum). Moreover, the half-life of fungicides under the influence of biostimulators was evaluated. Titanium caused most efficient mitigation of fungicide-induced toxicity reflected by greater increase of protein (104.80 mg g-1), carbohydrates (43.70 mg g-1), phenolic compounds (1023.18 µg g-1), and peroxidase activity (0.151 U mg-1) compared to treatment with silicon. The application of spiroketalamine and triazole fungicides with silicon decreased the activity of acid phosphatases (35.70 µM h-1 g-1) while inducing amino acid (861.40 µg kg-1) and catalase activity (1.056 U mg-1). Moreover, silicon was most effective in the reduction of mycotoxin contamination in wheat grain (61.17 µg g-1). This study indicated a shortened dissipation of propiconazole, spiroxamine, tebuconazole and triadimenol under the influence of titanium or silicon (DT50 = 1.85-4.82), which can therefore affect the grace period of wheat. The results of this comprehensive investigation demonstrated that titanium and silicon have a beneficial influence on wheat biochemistry and mitigate fungicide toxicity, which makes them widely suitable for optimizing plant health.

Effects of agricultural landscape structure, insecticide residues, and pollen diversity on the life-history traits of the red mason bee Osmia bicornis.

Agricultural landscapes have changed substantially in recent decades, shifting from the dominance of small fields (S) with diverse cropping systems toward large-scale monoculture (L), where landscape heterogeneity disappears. In this study, artificial nests of the red mason bee, Osmia bicornis, were placed in S and L landscape types on the perimeter of oilseed rape fields representing different oilseed rape coverages (ORC, % land cover). The local landscape structure around each nest was characterised within a 100, 200, 500, and 1000 m radius using ORC and 14 landscape characteristics, which were then reduced by non-metric multidimensional scaling (nMDS) to two axes: nMDS1 characterised the dataset primarily according to land fragmentation and the main crop, whereas nMDS2 captured the prevalence of more natural areas in the landscape. Pollen diversity and insecticide risk levels in the pollen provisions collected by the bees were analysed, and their dependence on the landscape structure was tested. Thereafter, the effects of pollen diversity, insecticide risk, and landscape structure on the life-history traits of bees and their sensitivity to topically applied Dursban 480 EC were determined. Pollen taxa richness in a single nest ranged from 3 to 12, and 34 pesticides were detected in the pollen at concentrations of up to 320 ng/g for desmedipham. The O. bicornis foraging range was relatively large, indicating that the landscape structure within a radius of ~1000 m around the nest is important for this species. Pollen diversity in the studied areas was of minor importance for bee performance, but the ORC or landscape structure significantly affected the life-history traits of the bees. Contamination of pollen with insecticides affected the bees by decreasing the mass of newly emerged adults but their sensitivity to Dursban 480 EC was not related to environmental variables.

Synthesis, characterization and biological activity of bifunctional ionic liquids based on dodine ion.

BACKGROUND: Development of new plant protection strategies has become an urgent matter in modern agriculture, in view of the evidently proved negative effect of currently used active ingredients of pesticides. In recent years, much effort has been made to eliminate the use of pesticides established to be toxic to pollinators. RESULTS: In this study, we present a group of new bifunctional ionic liquids based on dodine (N-dodecylguanidine) cation whose physical and biological properties have been modified relative to those of the commercially available N-dodecylguanidine acetate. The decreased level of residue of active substances in plant tissues reduces their availability to pollinators, which increases the safety of their use. Moreover, lower environmental impact in combination with high antifungal activity and an additional biological function, that is the systemic acquired resistance induction, are in line with the goals of sustainable agriculture. CONCLUSION: The presented approach shows the possibility of derivatization of commonly used fungicide into the form of bifunctional salts whose physical and biological properties can be easily modified. The paper reports successful design and synthesis of new sustainable and green chemicals for the modern agriculture, being less toxic to the environment and human health but still effective against pathogens. © 2021 Society of Chemical Industry.

Prokineticin 1-prokineticin receptor 1 signaling in trophoblast promotes embryo implantation and placenta development.

Successful pregnancy establishment in mammals depends on proper embryo-maternal communication. Prokineticin 1 (PROK1) is a secretory protein that exerts pleiotropic functions in various tissues. Despite the studies that have primarily been performed with human cell lines and mice, the function of PROK1 in trophoblasts has still not been fully elucidated. Hence, the aim of this study was to establish the role of PROK1 in trophoblasts during implantation and placentation. Prokineticin 1 mRNA was elevated in porcine trophoblasts during implantation and the early placentation period. Furthermore, we reveal that PROK1-PROKR1 signaling induces the expression of genes involved in the regulation of angiogenesis, immunological response, trophoblast cell adhesion, invasion, and proliferation, as well as stimulating phosphorylation of MAPK and PTK2. Ingenuity Pathway Analysis identified the aforementioned and also other functions associated with PROK1-regulated genes/proteins, such as cell-to-cell contact, epithelial tissue differentiation, Ca2+ release, lipid synthesis, and chemotaxis. We also showed evidence that PROK1 acting via PROKR1 increased trophoblast cell proliferation and adhesion. The PROK1-stimulated cell proliferation was mediated by PI3K/AKT/mTOR, MAPK, and cAMP, whereas adhesion was mediated by MAPK and/or PI3K/AKT signaling pathways. Concluding, our study suggests that PROK1 plays a pleiotropic role in trophoblast function during implantation and early placentation.

Health risk assessment of exposure to toxic elements resulting from consumption of dried wild-grown mushrooms available for sale.

Mushrooms exhibit a high ability to accumulate potentially toxic elements. The legal regulations in force in the European Union countries do not define the maximum content of elements in dried wild-grown mushrooms. This study presents the content of mercury (Hg), lead (Pb), cadmium (Cd) and arsenic (As) determined in dried wild-grown mushrooms (Boletus edulis and Xerocomus badius) available for sale. Moreover, the health risk associated with their consumption is assessed. The inductively coupled plasma mass spectrometry (Cd, Pb, As) and atomic absorption spectrometry (Hg) were used. The mean Hg, Cd, Pb and As concentration in Boletus edulis was 3.039±1.092, 1.983±1.145, 1.156±1.049 and 0.897±0.469 mg/kg and in Xerocomus badius 0.102±0.020, 1.154±0.596, 0.928±1.810 and 0.278±0.108 mg/kg, respectively. The maximum value of the hazard index (HI) showed that the consumption of a standard portion of dried Boletus edulis may have negative consequences for health and corresponded to 76.2%, 34.1%, 33% and 4.3% of the maximum daily doses of Hg, Cd, Pb and As, respectively. The results indicate that the content of toxic elements in dried wild-grown mushrooms should be monitored. The issue constitutes a legal niche where unfavourable EU regulations may pose a threat to food safety and consumer health.

Impact of broad-spectrum pesticides used in the agricultural and forestry sector on the pesticide profile in wild boar, roe deer and deer and risk assessment for venison consumers.

The present work is the first extensive study of large-scale pesticides research in wild animals. The investigation covered three game species: wild boar (n = 42), roe deer (n = 79) and deer (n = 15) collected from north-eastern Poland. To characterize the 480 pesticides in muscle samples, LC-GC-MS/MS techniques were used. A total of 28 compounds were detected: 5 neonicotinoids, 6 organochlorine and 5 other insecticides, 9 fungicides and 4 herbicides, in the range of 0.1-85.3 ng g-1. Over four hundred detections were done. The highest mean concentrations were as follows: anthraquinone (85.3 ng g-1) > DDT-p,p' (4.6 ng g-1) > imidacloprid (4.3 ng g-1) > permethrin (3.6 ng g-1) > thiacloprid (2.8 ng g-1). DDT and metabolites were the most frequently detected, followed by acetamiprid, tebuconazole, clothianidin and imidacloprid. Overall, 92% samples with residues were recorded, including 100% of wild boar, 88% of roe deer and 86% of deer. More than one pesticide (up to 9) was found in over 73% of the tested samples. The estimated chronic and acute risk to consumers of venison were very low (below 1% ADI and ARfD). This interdisciplinary study may be helpful for estimating ecological risk to wild animals and risk to consumers of wild animal products, and also as a source of biomonitoring data.

A global study of pesticides in bees: QuEChERS as a sample preparation methodology for their analysis - Critical review and perspective.

To this day, it remains unknown what the cause of decline of honey bee populations is and how to prevent this phenomenon efficiently. Poisonings with pesticides are assumed to be among the main causes for the decline of the honey bee population. Despite the significant progress observed in analytics over recent years, research aimed at improving methods applied in diagnostics of bee poisoning is still in progress. This is no easy task, since determination of the content of trace amounts (often equal to sublethal doses) of a wide range of compounds with diverse physico-chemical properties in honey bee samples with a complex matrix composition poses a serious challenge to modern analytics. This overview is the first to include a comprehensive critical assessment of analytical methods proposed for quantification of pesticides in honey bees over the last decade. Since the QuEChERS method is currently of great significance to ensuring accurate and reliable results of pesticide quantification in honey bees, the present overview focuses on the major aspects of this method, which will provide a comprehensive reference for scientists. The review focuses on the limitations of methods and on potential future prospects. It also contains information on the detection of pesticides in honey bees between 2010 and 2020 and characterizes the pesticide classes which are most toxic to these insects. This is extremely important, not just in the context of understanding the potential adverse impact of pesticides, manifesting as losses in bee colonies; it is also intended to facilitate decision-making in future research related to this difficult yet very important subject.

Exposure of wild boars (Sus scrofa L) to neonicotinoid insecticides.

The aim was to determine, for the first time, concentrations of 7 neonicotinoids (NEOs) and 5 metabolites in Sus scrofa from hunting areas in north-eastern Poland and assess the risk to consumers eating boar meat. 42 wild boar muscle samples were collected over a one-year period. The concentrations of 12 NEOs were determined by a fully validated LC-ESI-MS/MS protocol based on ultrasonic, freezing and cleanup EMR-lipid sample preparation. NEOs were present in over 83% of samples, 17% had no residue, and one pesticide was present in 36% of samples. Most often found were: clothianidin (35%), acetamiprid and imidacloprid (33%), thiacloprid (31%), thiamethoxam (9%), and the average concentrations were (ng g-1): thiacloprid 6.2 > imidacloprid 5.7 > acetamiprid 4.6 > clothianidin 2.2 > thiacloprid 1.6 > thiamethoxam 1.0. Multi-residue samples were found, one with 7 and one with 5 NEOs. Two NEOs were present in 24%; 3 in 39% and 4 in 10% of samples. In the metabolic degradation of acetamiprid, imidacloprid and thiacloprid, it was observed that metabolites account for no more than 8.5% of the measured parent substance. Acetamiprid-n-desmethyl was noted most often (21%). Due to the detection of NEOs in a large proportion of samples, chronic and acute risk assessment were performed. The estimated chronic and acute risk for consumers from NEOs neonicotinoids through the consumption of wild boar was very low and amounted to respectively 0.02% of ADI and 0.86% of ARfD.

Estradiol-17ß Regulates Expression of Luteal DNA Methyltransferases and Genes Involved in the Porcine Corpus Luteum Function In Vivo.

The corpus luteum (CL) is a temporary endocrine gland vital for pregnancy establishment and maintenance. Estradiol-17ß (E2) is the major embryonic signal in pigs supporting the CL's function. The mechanisms of the luteoprotective action of E2 are still unclear. The present study aimed to determine the effect of E2 on luteal expression of factors involved in CL function. An in vivo model of intrauterine E2 infusions was applied. Gilts on day 12 of pregnancy and the estrous cycle were used as referential groups. Concentrations of E2 and progesterone were elevated in CLs of gilts receiving E2 infusions, compared to placebo-treated gilts. Estradiol-17ß stimulated luteal expression of DNA-methyltransferase 1 (DNMT1), but decreased expression of DNMT3B gene and protein, as well as DNMT3A protein. Similar results for DNMT3A and 3B were observed in CLs on day 12 of pregnancy compared to day 12 of the estrous cycle. Intrauterine infusions of E2 altered luteal expression of the genes involved in CL function: PTGFR, PTGES, STAR, HSD17B1, CYP19A1, and PGRMC1. Our findings indicate a role for E2 in expression regulation of factors related to CL function and a novel potential for E2 to regulate DNA methylation as putative physiological mechanisms controlling luteal gene expression.

Dissipation kinetics and processing behavior of boscalid and pyraclostrobin in greenhouse dill plant (Anethum graveolens L.) and soil.

BACKGROUND: Determining the environmental behavior and fate of chemical compounds during technological processing of plants is a task of great significance nowadays. However, the current knowledge is still incomplete for unique herbal matrices belonging to minor crops. The research in this article presents, for the first time, the dissipation kinetics and processing behavior of carboxamide boscalid (BOS) and stobilurin pyraclostrobin (PYR) fungicides during glasshouse dill (Anethum graveolens L.) cultivation. RESULTS: The half-lives (t1/2 ) of BOS and PYR after application at the recommended and double dosage were in the range: 1.62-2.01 days in plant and 2.08-4.85 days in soil, respectively. The processing behavior in dill was estimated after washing, hot air drying and drying in sunlight without/with pretreatment. Processing factors (PFs) were above 1 after drying (PF = 1.24-1.39 hot air; PF = 1.15-1.28 sunlight) and below this value when the washing step was applied (PF = 0.31-0.42 hot air; PF = 0.21-0.34 sunlight), indicating the highest effectiveness of reduction, up to 73% BOS and 79% PYR. CONCLUSION: BOS/PYR residues at pre-harvest intervals after both doses were below European Union (EU) maximum residue limits (MRLs). The highest effectiveness was noted for drying carried out with the washing step, which has a great influence on the concentration of residues in the final product. The findings can supplement PF databases not set for minor crops and can be used to establish MRLs and determine human exposures more accurately in risk assessment studies. © 2021 Society of Chemical Industry.

Synergistic action of estradiol and PGE2 on endometrial transcriptome in vivo resembles pregnancy effects better than estradiol alone†.

Successful pregnancy establishment in mammals depends on numerous interactions between embryos and the maternal organism. Estradiol-17ß (E2) is the primary embryonic signal in the pig, and its importance has been questioned recently. However, E2 is not the only molecule of embryonic origin. In pigs, prostaglandin E2 (PGE2) is abundantly synthesized and secreted by conceptuses and endometrium. The present study aimed to determine the role of PGE2 and its simultaneous action with E2 in changes in porcine endometrial transcriptome during pregnancy establishment. The effects of PGE2 and PGE2 acting with E2 were studied using an in vivo model of intrauterine hormone infusions, and were compared to the effects of E2 alone and conceptuses' presence on day 12 of pregnancy. The endometrial transcriptome was profiled using gene expression microarrays followed by statistical analyses. Downstream analyses were performed using bioinformatics tools. Differential expression of selected genes was verified by quantitative polymerase chain reaction. Microarray analysis revealed 2413 differentially expressed genes (DEGs) in the endometrium treated simultaneously with PGE2 and E2 (P < 0.01). No significant effect of PGE2 administered alone on endometrial transcriptome was detected. Gene ontology annotations enriched for DEGs were related to multiple processes such as: focal adhesion, vascularization, cell migration and proliferation, glucose metabolism, tissue remodeling, and activation of immune response. Simultaneous administration of E2 and PGE2 induced more changes within endometrial transcriptome characteristic to pregnancy than infusion of E2 alone. The present findings suggest that synergistic action of estradiol-17ß and PGE2 resembles the effects of pregnancy on endometrial transcriptome better than E2 alone.