Exposure to the extremely low-frequency electromagnetic field induces changes in the epigenetic regulation of gene expression in the endometrium.

Increasing technological development results in more sources of the extremely low-frequency electromagnetic field (ELF-EMF), which is recognized as an environmental risk factor. The results of the past study indicate that the ELF-EMF can affect the level of DNA methylation. The study aimed to determine whether the ELF-EMF induces changes in epigenetic regulation of gene expression in the endometrium of pigs during the peri-implantation period. Endometrial slices (100 ± 5 mg) collected on days 15-16 of pregnancy were exposed in vitro to the ELF-EMF at a frequency of 50 Hz for 2 h of treatment duration. To determine the impact of the ELF-EMF on elements of epigenetic regulations involved in DNA methylation, histone modification, and microRNA biogenesis in the endometrium, the DNMT1 and DNMT3a; EZH2, UHRF1, and MBD1; DICER1 and DGCR8 mRNA transcript and protein abundance were analyzed using Real-Time PCR and Western blot, respectively. Moreover, EED and SUZ12 mRNA transcript, global DNA methylation, and the activity of histone deacetylase (HDAC) were analyzed. The changes in the abundance of DNMT1 and DNMT3a, EZH2 mRNA transcript and protein, EED and SUZ12 mRNA transcript, global DNA methylation level, HDAC activity, and the abundance of proteins involved in microRNA biogenesis evoked by the ELF-EMF in the endometrium were observed. The ELF-EMF possesses the potential to alter epigenetic regulation of gene expression in the porcine endometrium. Observed alterations may be the reason for changes in the transcriptomic profile of the endometrium exposed to the ELF-EMF which in turn may disrupt biological processes in the uterus during peri-implantation.

Extremely low-frequency electromagnetic field exposure alters DNA methylation levels in the endometrium of pigs during the peri-implantation period.

CONTEXT: Extremely low-frequency electromagnetic field (ELF-EMF) emission is increasing due to substantial technological progress. The results of previous research provided evidence that ELF-EMF may exert changes in molecular mechanisms that control female reproduction. AIMS: We hypothesised that short-term ELF-EMF treatment alters the DNA methylation level of genes in the endometrium. Hence, the research aimed to determine the methylation level of selected genes whose expression was altered in response to ELF-EMF radiation in the endometrium of pigs during the peri-implantation period (days 15-16 of pregnancy). METHODS: Porcine endometrial slices (100±5mg) were collected during the peri-implantation period and exposed to ELF-EMF at a frequency of 50Hz for 2h in vitro . The control endometrium was not exposed to ELF-EMF. The level of DNA methylation in the promoter regions of EGR2 , HSD17B2 , ID2 , IL1RAP, MRAP2, NOS3, PTGER4, SERPINE1, VDR and ZFP57 was tested using qMS-PCR. KEY RESULTS: In the endometrium exposed to ELF-EMF, the level of methylation of HSD17B2 , MRAP2 , SERPINE1, VDR and ZFP57 was not altered; the level of methylation of EGR2 , ID2 and PTGER4 increased, and the level of methylation of IL1RAP and NOS3 decreased. CONCLUSIONS: ELF-EMF may alter the level of DNA methylation in the endometrium during the peri-implantation period. IMPLICATIONS: Changes in the DNA methylation induced by ELF-EMF may affect the transcriptomic profile of the endometrium and disturb physiological processes accompanying implantation and embryo development.

Extremely low-frequency electromagnetic field (ELF-EMF) induces alterations in epigenetic regulation in the myometrium - An in vitro study.

Previous research by the authors indicated that an extremely low-frequency electromagnetic field (ELF-EMF) evokes molecular alterations in the porcine myometrium. It was hypothesized that the ELF-EMF could induce alterations in the epigenetic regulation of gene expression in the myometrium. In the current study, slices of the porcine myometrium during the peri-implantation period (n = 4) were used for further in vitro exposition to ELF-EMF (50 Hz, 8 mT, 2 h treatment duration). The study tested whether the ELF-EMF may affect: 1/the expression of DNA (cytosine-5)-methyltransferase 1 (DNMT1) and DNA (cytosine-5)-methyltransferase 3a (DNMT3a), 2/the level of genomic DNA methylation, and 3/the level of amplification of methylated and unmethylated variants of promoter regions of selected genes with altered expression in response to ELF-EMF. It was found that ELF-EMF treatment increased DNMT1, decreased DNMT3a mRNA transcript and protein abundance, and increased the level of genomic DNA methylation. The direction of alterations in the level of amplification of methylated and unmethylated variants of the promoter region of selected genes with altered expression, i.e. prodynorphin (PDYN), interleukin 15 (IL15) signal transducer and activator of transcription 5A (STAT5A), tumor necrosis factor (TNF), and between down-regulated genes were early growth response 2 (EGR2), hyaluronan and proteoglycan link protein 1 (HAPLN1), and uteroferrin associated basic protein-2 (UABP2), mostly involving the direction of changes in their transcriptional activity, which was evaluated in a previous study by the authors. Thus, ELF-EMF radiation disturbs epigenetic mechanisms, which may underlay ELF-EMF-related transcriptomic alterations in the myometrium.

The effect of electromagnetic field (EMF) exposure on synthesis and release of steroid hormones by the porcine conceptuses during the peri-implantation period.

CONTEXT: Electrical devices and power systems are the sources of EM-waves which propagate everywhere in the environment. AIMS: The study aimed to determine whether EMF induced changes in the steroidogenesis of conceptuses and whether progesterone (P4 ) may be a possible protectant against the effects of EMF radiation. METHODS: The entire porcine conceptuses were collected during the peri-implantation period (days 15-16 of pregnancy), divided into fragments (100mg) and treated in vitro with EMF (50Hz or 120Hz, 2 or 4h exposure), and examined to determine of CYP17A1 , HSD3B1 , CYP19A3 , and HSD17B4 mRNA transcript and encoded protein abundance and the release of steroid hormones. Selected fragments of conceptuses were treated with P4 . KEY RESULTS: In conceptuses incubated without P4 , EMF at 120Hz decreased androstenedione (A4 ) and testosterone (T) release after 2h and increased oestrone (E1 ) release at 50Hz and 120Hz after 4h exposure. In P4 -treated conceptuses, EMF (50 and 120Hz, 4h exposure) decreased CYP19A3 mRNA transcript abundance, and increased (120Hz, 2h exposure) oestradiol-17ß (E2 ) release. CONCLUSIONS: The EMF radiation alters androgen and oestrogen synthesis and release from the conceptuses of pigs during the peri-implantation period. The P4 exerts protective effects on androgens and E1 release but it sensitises the conceptuses when comes to the mechanism of oestrogen synthesis and release during EMF radiation. IMPLICATIONS: The effect of EMF radiation on the steroidogenic pathway in conceptuses may induce disturbances in their proper development and implantation.

Electromagnetic field exposure alters in vitro estrogen biosynthesis and its release by the porcine endometrium in the peri-implantation period.

The electromagnetic field (EMF) is an environmental risk factor that may impair living organisms. This study aims to determine the functional effects of EMF exposure at 50 and 120 Hz for 2 or 4 h on estrogen synthesis and release in the endometrium. Endometrial slices were isolated from pigs (n = 5) during the peri-implantation period. To check whether progesterone (P4) exerts any protective effects against EMF, selected EMF-treated slices were also treated with P4. CYP19A3 mRNA transcript abundance was higher in slices exposed to EMF (50 Hz, 4 h) and treated with P4. HSD17B4 mRNA transcript abundance was higher in slices exposed to EMF (50 and 120 Hz, 2 h) without P4 treatment. Both EMF (50 Hz, 2 h) and EMF (50 and 120 Hz, 4 h) increased HSD17B4 mRNA transcript abundance in the presence of P4; EMF (120 Hz, 2 h, and 50 Hz, 4 h) decreased cytochrome P-450arom protein abundance in tissue slices not treated with P4. Under exposure to EMF at 120 Hz (2 h), the abundance of hydroxysteroid 17ß dehydrogenase decreased in P4-treated slices and increased in slices not treated with P4 (4 h). Progesterone treatment decreased the release of estradiol-17ß (E2) in endometrial slices exposed to EMF at 50 Hz (2 h), whereas in slices not treated with P4, EMF (120 Hz, 2 h) increased estrone (E1) release compared to control (without EMF). The EMF could disrupt the synthesis and release of E1 and E2 by the porcine endometrium during the peri-implantation period.

Profiles of posttraumatic stress disorder (PTSD), complex posttraumatic stress disorder (CPTSD), and subjective well-being in a sample of adult children of alcoholics.

OBJECTIVE: The main aim of this study was to examine the heterogeneity of a sample of adult children of alcoholics (ACOAs) within the International Classification of Diseases (ICD)-11 posttraumatic stress disorder (PTSD) and complex PTSD (CPTSD) as well as DSM-5 PTSD diagnoses regarding the participants' subjective well-being (SWB). In addition, the construct validity of the ICD-11 CPTSD was assessed, and the ICD-11 and DSM-5 PTSD diagnoses among participants were compared. METHOD: The sample consisted of 609 ACOAs. Participants filled out the PTSD Checklist for the DSM-5 (PCL-5), International Trauma Questionnaire (ITQ), Satisfaction with Life Scale (SWLS), Positive and Negative Affect Schedule (PANAS-X), and the Children of Alcoholics Screening test (CAST-6). RESULTS: We observed many fewer PTSD cases when we followed the ICD-11 criteria compared to cases of PTSD diagnosed based on DSM-5 criteria among participants. In addition, latent profile analysis (LPA) did not provide evidence of the construct validity of CPTSD. Finally, we found heterogeneity of ACOAs sample in terms of PTSD/CPTSD profiles, which were differently related to their SWB. CONCLUSIONS: Further research is needed to establish to what extent CPTSD and PTSD are separate and discrete disorders and why such discrepancy in PTSD level is observed when we follow either DSM-5 or ICD-11. In addition, the person-centered approach may provide more insight than variable-centered methods and thus, undermine the "uniformity myths" about ACOAs. (PsycInfo Database Record (c) 2022 APA, all rights reserved).

Nutritional restriction during the peri-conceptional period alters the myometrial transcriptome during the peri-implantation period.

This study hypothesized that female peri-conceptional undernutrition evokes transcriptomic alterations in the pig myometrium during the peri-implantation period. Myometrium was collected on days 15-16 of pregnancy from pigs fed a normal- (n = 4) or restricted-diet (n = 4) from conception until day 9th of pregnancy, and the transcriptomic profiles of the tissue were compared using Porcine (V2) Expression Microarrays 4 × 44 K. In restricted diet-fed pigs, 1021 differentially expressed genes (DEGs) with fold change ≥ 1.5, P ≤ 0.05 were revealed, and 708 of them were up-regulated. Based on the count score, the top within GOs was GO cellular components "extracellular exosome", and the top KEGG pathway was the metabolic pathway. Ten selected DEGs, i.e. hydroxysteroid (17ß) dehydrogenase 8, cyclooxygenase 2, prostaglandin F receptor, progesterone receptor membrane component 1, progesterone receptor membrane component 2, annexin A2, homeobox A10, S-phase cyclin A-associated protein in the ER, SRC proto-oncogene, non-receptor tyrosine kinase, and proliferating cell nuclear antigen were conducted through qPCR to validate microarray data. In conclusion, dietary restriction during the peri-conceptional period causes alterations in the expression of genes encoding proteins involved i.a. in the endocrine activity of the myometrium, embryo-maternal interactions, and mechanisms regulating cell cycle and proliferation.

Effect of the Electromagnetic Field (EMF) Radiation on Transcriptomic Profile of Pig Myometrium during the Peri-Implantation Period-An In Vitro Study.

The electromagnetic field (EMF) affects the physiological processes in mammals, but the molecular background of the observed alterations remains not well established. In this study was tested the effect of short duration (2 h) of the EMF treatment (50 Hz, 8 mT) on global transcriptomic alterations in the myometrium of pigs during the peri-implantation period using next-generation sequencing. As a result, the EMF treatment affected the expression of 215 transcript active regions (TARs), and among them, the assigned gene protein-coding biotype possessed 90 ones (differentially expressed genes, DEGs), categorized mostly to gene ontology terms connected with defense and immune responses, and secretion and export. Evaluated DEGs enrich the KEGG TNF signaling pathway, and regulation of IFNA signaling and interferon-alpha/beta signaling REACTOME pathways. There were evaluated 12 differentially expressed long non-coding RNAs (DE-lnc-RNAs) and 182 predicted single nucleotide variants (SNVs) substitutions within RNA editing sites. In conclusion, the EMF treatment in the myometrium collected during the peri-implantation period affects the expression of genes involved in defense and immune responses. The study also gives new insight into the mechanisms of the EMF action in the regulation of the transcriptomic profile through lnc-RNAs and SNVs.

Electromagnetic Field (EMF) Radiation Alters Estrogen Release from the Pig Myometrium during the Peri-Implantation Period.

An electromagnetic field (EMF) may affect the functions of uterine tissues. This study hypothesized that EMF changes the estrogenic activity of pig myometrium during the peri-implantation period. Tissue was collected on days 15-16 of the gestation and incubated in the presence of EMF (50 and 120 Hz, 2 and 4 h). The cytochrome P450 aromatase type 3 (CYP19A3) and hydroxysteroid 17ß dehydrogenase type 4 (HSD17B4) mRNA transcript abundance, cytochrome P450arom (aromatase), and 17ß hydroxysteroid dehydrogenase 17ßHSD) protein abundance and estrone (E1) and estradiol-17ß (E2) release were examined using Real-Time PCR, Western blot and radioimmunoassay. Selected myometrial slices were treated with progesterone (P4) to determine whether it functions as a protector against EMF. CYP19A3 mRNA transcript abundance in slices treated with EMF was less at 50 Hz (2 h) and greater at 120 Hz (2 and 4 h). HSD17B4 mRNA transcript was greater in slices treated with EMF at 120 Hz (2 h). Progesterone diminished EMF-related effects on CYP19A3 and HSD17B4. When P4 was added, EMF had suppressive (50 and 120 Hz, 2 h) or enhancing (50 Hz, 4 h) effects on aromatase abundance. The E1 release was lower after 4 h of EMF treatment at 50 Hz and P4 did not protect myometrial E1 release. In conclusion, EMF alters the synthesis and release of E1 and did not affect E2 release in the myometrium during the peri-implantation period.

Proteomic profile alterations in porcine conceptuses during early stages of development.

The dynamic embryo development during the early stages of gestation requires precise molecular changes, including proteomic ones. We aimed to find unique proteins for porcine conceptuses specifically during the peri-implantation period, i.e. on days 15-16 of pregnancy. The proteomic profile of these conceptuses was compared with conceptuses at an earlier stage of the development, i.e. collected during maternal recognition of pregnancy on days 12-13 of pregnancy. The 2DE, gel image analysis, and MALDI TOF mass spectrometry were used 500 protein spots were annotated as common to conceptuses harvested during both studied periods. Proteomic profile of the conceptuses collected during the peri-implantation period contains 24 unique proteins. Identified unique for the peri-implantation period proteins are involved in adhesion processes, cadherin, and actin-binding, and actin filament organization, extracellular matrix organization, and cytoskeleton organization. Systemic analysis of identified proteins confirmed their involvement in cell adhesion and cytoskeletal organization as being two major affected functions. The unique proteins might be recognized as factors conditioning the proper peri-implantation embryo development and gaining competences for implantation. In further studies, BRCA1 might be considered as a candidate for a potential marker of embryonic competences for implantation in pigs.

Effects of electromagnetic field (EMF) radiation on androgen synthesis and release from the pig endometrium during the fetal peri-implantation period.

An electromagnetic field (EMF) may have effects on female reproduction. This study was conducted to determine whether EMF [50 and 120 Hz, 2 and 4 h of incubation in the presence or absence of progesterone (P4, 10-5 M)] affects androgen synthesis and release from the pig endometrium. Endometrial slices were collected from pigs (n = 5) during the fetal peri-implantation period (i.e., days 15-16 of gestation) and treated in vitro with EMF. The selected endometrial slices were treated with P4 to determine whether this hormone has effects on protection of the tissue from EMF radiation. The CYP17A1 and HSD3B1 mRNA transcript abundance, steroid 17αhydroxylase/17, 20-lyase (cytochrome P450c17) and hydroxyΔ5steroid dehydrogenase/3ß and steroidΔisomerase (3ßHSD) protein abundance were examined using Real-Time PCR and Western Blot procedures, respectively. In media collected after incubation, the concentrations of androstenedione (A4) and testosterone (T) were quantified used a RIA. When P4 was added to the culture medium, EMF radiation had suppressive effects on endometrial T release after 2 and 4 h of incubation when the EMF treatment was occurring and increased A4 release after 4 h of incubation with EMF at 120 Hz. When there was no inclusion of P4, release of A4 was decreased after 2 h of EMF treatment at 120 Hz and after 4 h of EMF treatment at 50 and 120 Hz. Progesterone did not have functions that protected the pig endometrium against EMF radiation during the fetal peri-implantation period.

Effects of LH and FSH on androgen and oestrogen release in the myometrium of pigs during the oestrous cycle and early pregnancy.

The porcine myometrium possesses steroidogenic activity. LH and FSH are hypothesised to regulate the myometrial production of androstenedione (A4), testosterone (T), oestrone (E1) and 17ß-oestradiol (E2). In this study, we used myometrium collected from cycling (n=15) and pregnant (n=15) pigs on Days 10-11, 12-13 and 15-16 of the oestrous cycle or pregnancy to determine: (1) the abundance of LH and FSH receptor (LH/choriogonadotrophin receptor (CGR) and FSHR) mRNA and protein; (2) activity of 17ß-hydroxysteroid dehydrogenase 1 (17ßHSD1); and (3) A4, T, E1 and E2 release in response to LH and FSH treatment, used at doses 10 or 100ng mL-1 for 6h. In results, the myometrium possesses LH/CGR and FSHR with minor alterations in their expression in the course of the oestrous cycle or early pregnancy. 17ßHSD1 activity was the highest on Days 12-13 of the oestrous cycle and the lowest on Days 15-16 of the oestrus cycle and pregnancy, when compared to the other studied days of the oestrous cycle or pregnancy. The LH and FSH treatment increased A4 release on Days 12-13 of the oestrous cycle, and E1 and E2 release on Days 15-16 of the oestrous cycle. Moreover, on Days 12-13 E2 release was increased in response to FSH treatment (100ng mL-1) in cycling pigs and in response to LH (100ng mL-1) in pregnant pigs. In conclusion, the myometrium of pregnant and non-pregnant pigs expresses LH/CGR and FSHR and has 17ßHSD1 activity. In addition, the amount of A4, E1, and E2 release from the myometrium is altered in response to LH and FSH, especially in cycling pigs. LH and FSH appear to be important regulators of myometrial oestrogen release in pigs mostly during luteolysis.

Consequences of electromagnetic field (EMF) radiation during early pregnancy - androgen synthesis and release from the myometrium of pigs in vitro.

An electromagnetic field (EMF) has been found to affect reproductive processes in females. The aim of this study was to determine the effect of low, non-ionizing EMF radiation on the steroidogenic activity of myometrium collected from pigs during the fetal peri-implantation period. Myometrial slices were treated with an EMF (50 and 120 Hz, 2 and 4 h of incubation) and examined for the aromatase cytochrome P450 17α-hydroxylase/C17-20lyase (CYP17A1) and 3ß-hydroxysteroid dehydrogenase/Δ5-Δ4 isomerase (HSD3B1) mRNA transcript abundance, cytochrome P450c17 and 3ßHSD protein abundance and the secretion of androstenedione (A4) and testosterone (T). To determine whether progesterone (P4) functions as a protectant from EMF radiation, the selected slices were treated with P4. In slices incubated without P4, EMF at 50 Hz altered cytochrome P450c17 protein abundance (4 h), HSD3B1 mRNA transcript abundance (4 h) and A4 release (2 h) as well as T release (2 h) in P4-treated slices. The EMF at 120 Hz in non P4-treated slices altered A4 release (2 and 4 h) whereas in P4-treated slices altered CYP17A1 mRNA transcript abundance (4 h), 3ßHSD protein abundance (4 h), A4 (4 h) and T release (2 h). In conclusion, EMF radiation in the myometrium collected during the peri-implantation period alters the CYP17A1 and HSD3B1 mRNA transcript and encoded protein abundance, and androgen release due to the time of treatment and P4 presence or absence. The P4 did not function directly as an obvious protector against EMF radiation in the myometrium of pigs during the fetal peri-implantation period.

Expression of Insulin-Like Growth Factor 1 (IGF-1) and Epidermal Growth Factor (EGF) Receptors and the Effect of IGF-1 and EGF on Androgen and Estrogen Release in the Myometrium of Pigs-In Vitro Study.

Porcine myometrium possesses steroidogenic activity but its regulation is not well understood. It was hypothesized that the regulators of myometrial steroidogenesis are insulin-like growth factor 1 (IGF-1) and epidermal growth factor (EGF), which were found to modulate the steroidogenic activity of the endometrium and embryos. Myometrial slices were collected from gravid and nongravid pigs on days 10 or 11, 12 or 13 and 15 or 16 and studied for: (1) the relative abundance of IGF-1R and EGFR mRNA transcripts and proteins, to determine myometrial readiness to response growth factors treatment and (2) the effect of IGF-1 or EGF on the myometrial release of androstenedione (A4), testosterone (T), estrone (E1) and estradiol-17ß (E2). The results showed that the relative expression and abundance of IGF-1R and EGFR in the myometrium were altered regarding the female reproductive status. During the estrous cycle, EGF increased myometrial release of A4 on days 12-13 and E2 on days 15-16. In gravid pigs (days 15-16), IGF-1 and EGF increased the E1 release. In conclusion: (1) porcine myometrium possesses the potential to respond to IGF-1 and EGF treatment, (2) EGF significantly increases myometrial A4 and E2 release in cyclic pigs, while IGF-1 and EGF increase the E1 release in gravid pigs.

Allelopathic effects of Chara species (C. aspera, C. baltica, and C. canescens) on the bloom-forming picocyanobacterium Synechococcus sp.

The role of macroalgal allelopathy in aquatic systems has received increasing attention as a potential means of controlling cyanobacterial blooms. However, the allelopathic activity of Chara sp. on coexisting and bloom-forming picocyanobacteria is still largely unknown. Therefore, the laboratory experiments were conducted to investigate the allelopathic activity of extracts of Chara aspera, C. baltica, and C. canescens on the growth, the fluorescence parameters: maximum and effective quantum yield of photosystem II (PSII) photochemistry (Fv/Fm and ΦPSII, respectively) and photosynthesis parameters such as the initial slope of photosynthesis-irradiance (P-E) curves (alpha) and photosynthetic capacity (Pm) of the picocyanobacterium Synechococcus sp. Batch cultures of picocyanobacterium were exposed to three concentrations of extracts originating from three charophyte cultures and the effect was followed at three sampling times. Dried specimens of C. aspera, C. baltica, and C. canescens were extracted in the water-based matrix and the initial Synechococcus sp. inoculum, derived from unialgal culture media, was used. We found both negative and positive allelopathic effects of all tested Chara extracts on Synechococcus sp. The strongest adverse impact of picocyanobacterium growth was caused by C. baltica. This study clearly demonstrated that the allelopathic effect depends on the Chara species identity. Our results also suggested that some allelopathic Chara sp. have the potential to mitigate harmful cyanobacterial blooms in systems dominated by Synechococcus sp.