Bifunctional role of ephrin A1-Eph system in stimulating cell proliferation and protecting cells from cell death through the attenuation of ER stress and inflammatory responses in bovine mammary epithelial cells.

Structural and functional development of the mammary gland is constant in the mammary gland life cycle. Eph receptors and their ligands, ephrins, control events through cell-to-cell interactions during embryonic development, and adult tissue homeostasis; however, little information on participation of ephrin A1, a representative ligand of the Eph receptor, in the development and function of normal mammary glands is known. In this study, we demonstrated functional effects of the ephrin A1-Eph system and mechanisms of its action on bovine mammary epithelial (MAC-T) cells. The in vitro cultured MAC-T cells expressed the ephrin A1 ligand and EphA1, A2, A4, A7, and A8 among the eight members of the Eph A family. Our results revealed that ephrin A1 induced MAC-T cell cycle progression and stimulated cell proliferation with abundant expression of nucleic PCNA and cyclin D1 proteins. Additionally, ephrin A1 induced activation of intracellular signaling molecules involved in PI3 K/AKT and MAPK signaling, and the proliferation-stimulating effect of ephrin A1 was mediated by activation of these pathways. Furthermore, ephrin A1 influenced expression and activation of various ER stress-related proteins and protected MAC-T cells from stress-induced cell death. Finally, ephrin A1 alleviated LPS-induced cell death through down-regulation of inflammatory cytokines. In conclusion, the results of this study suggest that the Eph A-ephrin A1 system is a positive factor in the increase and maintenance of epithelial cells in mammary glands of cows; the signaling system contributes to development, remodeling, and functionality of normal mammary glands and could overcome mastitis in cows and other mammals.

Cysteine-X-cysteine motif chemokine ligand 12 and its receptor CXCR4: expression, regulation, and possible function at the maternal-conceptus interface during early pregnancy in pigs.

Cysteine-X-cysteine (CXC) motif chemokine ligand 12 (CXCL12) and its receptor, CXC chemokine receptor type 4 (CXCR4), are involved in regulating the proliferation, migration, and survival of trophoblast cells and the maternal immune response in humans and mice. The present study examined the expression, regulation, and function of CXCL12 and CXCR4 at the maternal-conceptus interface during pregnancy in pigs. The endometrium expressed CXCL12 and CXCR4 mRNAs with the greatest CXCL12 abundance on Day 15 of pregnancy. CXCL12 protein was localized mainly in endometrial epithelial cells, while CXCR4 protein was localized in subepithelial stromal cells, vascular endothelial cells, and immune cells in blood vessels in the endometrium during the estrous cycle and pregnancy. CXCL12 protein was detected in uterine flushing on Day 15 of pregnancy. The conceptus during early pregnancy and chorioallantoic tissues during mid-to-late pregnancy expressed CXCL12 and CXCR4. Interferon-γ increased the abundance of CXCL12, but not CXCR4 mRNA in endometrial explants. Recombinant CXCL12 (rCXCL12) protein dose-dependently increased migration of cultured porcine trophectoderm cells and peripheral blood mononuclear cells (PBMCs). Furthermore, rCXCL12 caused migration of T cells, but not natural killer cells, in PBMCs. This study revealed that interferon-γ-induced CXCL12 and its receptor, CXCR4, were expressed at the maternal-conceptus interface and increased the migration of trophectoderm cells and T cells at the time of implantation in pigs. These results suggest that CXCL12 may be critical for the establishment of pregnancy by regulating trophoblast migration and T cell recruitment into the endometrium during the implantation period in pigs.

Speckle noise reduction for digital holographic images using multi-scale convolutional neural networks.

In this Letter, we propose a fast speckle noise reduction method with only a single reconstructed image based on convolutional neural networks. The proposed network has multi-sized kernels that can capture the speckle noise component effectively from digital holographic images. For robust noise reduction performance, the network is trained with a large noisy image dataset that has object-dependent noise and a wide range of noise levels. The experimental results show the fast, robust, and outstanding speckle noise reduction performance of the proposed approach.

The impact of production of extended-spectrum ß-lactamases on the 28-day mortality rate of patients with Proteus mirabilis bacteremia in Korea.

BACKGROUND: The incidence of Proteus mirabilis antimicrobial resistance, especially that mediated by extended-spectrum ß-lactamases (ESBLs), has increased. We investigated the impact of ESBL production on the mortality of patients with P. mirabilis bacteremia in Korea. METHODS: Patients diagnosed with P. mirabilis bacteremia between November 2005 and December 2013 at a 2000-bed tertiary care center in South Korea were included in this study. Phenotypic and molecular analyses were performed to assess ESBL expression. Characteristics and treatment outcomes were investigated among ESBL-producing and non-ESBL-producing P. mirabilis bacteremia groups. A multivariate analysis of 28-day mortality rates was performed to evaluate the independent impact of ESBLs. RESULTS: Among 62 P. mirabilis isolates from 62 patients, 14 expressed ESBLs (CTX-M, 2; TEM, 5; both, 6; other, 1), and the 28-day mortality rate of the 62 patients was 17.74%. No clinical factor was significantly associated with ESBL production. The 28-day mortality rate in the ESBL-producing group was significantly higher than that in the non-ESBL-producing group (50% vs. 8.3%, p = 0.001). A multivariate analysis showed that ESBL production (odds ratio [OR], 11.53, 95% confidence interval [CI], 2.11-63.05, p = 0.005) was independently associated with the 28-day mortality rate in patients with P. mirabilis bacteremia. CONCLUSIONS: ESBL production is significantly associated with mortality in patients with bacteremia caused by P. mirabilis. Rapid detection of ESBL expression and prompt appropriate antimicrobial therapy are required to reduce mortality caused by P. mirabilis bacteremia.

Stem cell factor-induced AKT cell signaling pathway: Effects on porcine trophectoderm and uterine luminal epithelial cells.

Stem cell factor (SCF) is a multipotent growth factor that elicits diverse biological actions in various aspects of embryogenesis and animal development. The aim of the present study was to assess SCF-induced intracellular signaling and cellular activities in porcine trophectoderm (pTr) and uterine luminal epithelial (pLE) cells which are well known as useful to elucidate developmental events. SCF induced abundances of p-AKT, p-P70RSK and RPS6 proteins in pTr cells reached to their maximum, and then returned to basal levels by 120min. In pLE cells, SCF induced protracted effect to increase AKT phosphorylation which was well correlated with the time course for P70RSK and RPS6 phosphorylation. LY294002 (an inhibitor of AKT) decreased SCF-induced p-AKT, p-P70RSK and p-RPS6 proteins. Also, immunofluorescence analyses revealed that p-RPS6 was abundant within the cytoplasm of SCF-treated cells, but p-RPS6 was present only at basal levels in cells treated with LY294002. In the presence of LY294002, both SCF-stimulated transient and sustained AKT phosphorylation were inhibited in pLE cells. Furthermore, SCF increased migration of pTr and pLE cells, but LY294002 significantly reduced this effect of SCF. In conclusion, results of the present study suggest that SCF secreted by the endometrium induces autocrine/paracrine signaling responses that stimulate migration of pTr and pLE cells through activation of the AKT cell signaling pathway. Those results support the hypothesis that SCF is a critical regulatory factor for conceptus development and implantation during pregnancy in pigs.

Expression of hypoxia-inducible factor-1 by trophectoderm cells in response to hypoxia and epidermal growth factor.

The low oxygen environment in the uterine environment requires pre-implantation embryos to adapt to oxygen deficiency. Hypoxia-inducible factor (HIF)-1 is a master regulator whereby cells adapt to changes in oxygen concentrations. In addition to hypoxic conditions, non-hypoxic stimuli such as growth factors also activate expression of HIF-1. In this study, the mechanisms underlying low oxygen-dependent and epidermal growth factor (EGF)-dependent expression of HIF-1α were explored using porcine trophectoderm (pTr) cells. The results indicated that expression of HIF-1α and HIF-1ß mRNAs was not affected by low concentrations of oxygen; however, hypoxic conditions markedly increased the abundance of HIF-1α protein, especially in nuclei of pTr cells. Even under normoxic conditions, the abundance of HIF-1α protein increased in response to EGF. This EGF-mediated increase in HIF-1α protein was blocked through inhibition of translation by cycloheximide. The inhibitors LY294002 (PI3K-AKT inhibitor), U0126 (inhibitor of ERK1/2) and rapamycin (mTOR inhibitor) also blocked the ability of EGF to increase HIF-1α protein and to phosphorylate AKT, ERK1/2 and mTOR proteins. Both hypoxia and EGF induced proliferation of pTr cells. This ability of EGF to stimulate proliferation of pTr cells was suppressed by EGFR siRNA, but not HIF-1α siRNA, but a significant decrease in EGF-induced HIF-1α protein occurred when pTr cells were transfected with HIF-1α siRNA. The results of the present study suggest that pTr cells adapt to oxygen deficiency and proliferate in response to an oxygen-dependent HIF-1 system, and that EGF at maternal-conceptus interface can increase the abundance of HIF-1α protein via translational regulation through AKT, ERK1/2 and mTOR signaling cascades.

Lysophosphatidic Acid (LPA) Receptor 3-Mediated LPA Signal Transduction Pathways: A Possible Relationship with Early Development of Peri-Implantation Porcine Conceptus.

Lysophosphatidic acid (LPA) is a phospholipid with a variety of fatty acyl groups that mediates diverse biological effects on various types of cells through specific G protein-coupled receptors. LPA appears to play a significant role in many reproductive processes, including luteolysis, implantation, and placentation. Our previous study in pigs demonstrated that LPA and the LPA receptor system are present at the maternal-conceptus interface and that LPA increases uterine endometrial expression of prostaglandin-endoperoxide synthase 2 (PTGS2) through LPA receptor 3 (LPAR3). However, the role of LPA in conceptuses during early pregnancy has not been determined. Therefore, this study examined the effects of LPA in cell proliferation, migration, and activation of the intracellular signaling pathway in porcine conceptuses by using an established porcine trophectoderm (pTr) cell line isolated from Day 12 conceptuses. All examined LPA species with various fatty acid lengths increased proliferation and migration of pTr cells as the dosage increased. Immunoblot analyses found that LPA activated intracellular signaling molecules, extracellular signal-regulated kinase 1/2 (ERK1/2), ribosomal protein S6 kinase 90 kDa (P90RSK), ribosomal protein S6 (RPS6), and P38 in pTr cells. Furthermore, LPA increased expression of PTGS2 and urokinase-type plasminogen activator (PLAU), and the LPA-induced increases in PTGS2 and PLAU expression were inhibited by LPAR3 siRNA. Collectively, these results showed that LPA promotes proliferation, migration, and differentiation of pTr cells by activating the ERK1/2-P90RSK-RPS6 and P38 pathways, indicating that the LPA-LPAR3 system may be involved in the development of trophoblast during early pregnancy in pigs.

Mitogen activated protein kinase pathway-dependent effects of platelet-derived growth factor on migration of trophectoderm cells.

Successful development of the conceptus and implantation requires an intimate trophic connection between maternal uterus and conceptus mediated by local regulators including growth factors. Platelet-derived growth factor (PDGF) acts as a chemotactic factor for a variety of cell types. Current studies have determined that PDGF participates in rapid growth and development of cleavage stage embryos, but PDGF-induced effects on the growth and development of peri-implantation conceptus remains unknown. In the present study, PDGF induced phosphorylation of ERK1/2, AKT and RPS6 proteins in porcine trophectoderm (pTr) cells in a dose- and time-dependent manner. Addition of U0126 (an inhibitor of ERK1/2) or LY294002 (a PI3K inhibitor) blocked PDGF-induced effects on phosphorylation of signaling proteins. Combinations of PDGF and U0126 decreased PDGF-induced p-ERK1/2 and p-AKT1, but combinations of PDGF and LY294002 blocked only PDGF-induced AKT phosphorylation. Furthermore, PDGF significantly induced pTr cell migration and these stimulatory effects were blocked by U0126 and LY294002. Immunoreactive p-ERK1/2 and p-RPS6 proteins were abundant in pTr cells treated with PDGF, but U0126 reduced PDGF-induced p-ERK1/2 and p-RPS6 levels to basal amounts. Present study suggests that PDGF secreted into the maternal-conceptus microenvironment stimulates pTr cell migration through signal transduction cascades mediated by the ERK1/2 MAPK and AKT1 pathways.

Dietary cholesterol affects expression of prostatic acid phosphatase in reproductive organs of male rats.

Cholesterol homeostasis is strictly maintained to prevent abnormal biological processes that arise from excessive accumulation of cholesterol in tissues. Although dyslipidemia causes reproductive dysfunction and endocrine disruption in male rats, regulatory factors and mechanisms have not been clearly demonstrated. Therefore, the present study investigated the histology of male reproductive organs and the expression of prostatic acid phosphatase (also known as Acpp) that is secreted by cuboidal epithelium of the prostate gland in response to a normal diet and a high-cholesterol diet. The high cholesterol diet increased total cholesterol and low density lipoprotein (LDL) levels and decreased high density lipoprotein (HDL) levels. Histological analyses showed considerable alterations in the prostate indicating excessive papillary projections within the acinar lumen in response to the high cholesterol diet. In addition, Acpp expression was decreased in the penis of rats fed the high cholesterol diet and it was predominantly localized in the urethral epithelium and penile follicle that is precursor of penile spines. Moreover, Acpp was reduced slightly in the testes, but differential expression of Acpp in the prostate in response to dietary cholesterol was not detected. Furthermore, target microRNAs (miRs) of Acpp such as miR-192 and miR-215 regulated Acpp gene expression at the post-transcriptional levels by binding to specific sites within its 3'-UTR. These results indicate that Acpp plays an important role in growth and development of the penis of rats, and its expression is modulated at epigenomic levels via specific miRs.

Stimulatory Effect of Vascular Endothelial Growth Factor on Proliferation and Migration of Porcine Trophectoderm Cells and Their Regulation by the Phosphatidylinositol-3-Kinase-AKT and Mitogen-Activated Protein Kinase Cell Signaling Pathways.

Vascular endothelial growth factor (VEGF), a potent stimulator for angiogenesis, is likely to regulate implantation by stimulating endometrial angiogenesis and vascular permeability. In addition to known angiogenetic effects, VEGF has been suggested to participate in development of the early embryo as a mediator of fetal-maternal dialogue. Current studies have determined VEGF in terms of its role in endometrial vascular events, but VEGF-induced effects on the peri-implantation conceptus (embryo and extraembryonic membranes) remains unknown. In the present study, endometrial VEGF, VEGF receptor-1 (VEGFR-1), and VEGF receptor-2 (VEGFR-2) mRNAs increased significantly during the peri-implantation period of pregnancy as compared to the estrous cycle. Expression of VEGF, VEGFR-1, and VEGFR-2 mRNAs was abundant in endometrial luminal and glandular epithelia, endothelial blood vessels, and scattered cells in the stroma and conceptus trophectoderm. In addition, porcine trophectoderm (pTr) cells treated with VEGF exhibited increased abundance of phosphorylated (p)-AKT1, p-ERK1/2, p-p70RSK, p-RPS6, and p-4EBP1 in a time-dependent manner. The addition of U0126, an inhibitor of ERK1/2, inhibited VEGF-induced ERK1/2 phosphorylation, but AKT1 phosphorylation was not affected. The addition of LY294002, a PI3K inhibitor, decreased VEGF-induced phosphorylation of ERK1/2 and AKT1. Furthermore, VEGF significantly stimulated proliferation and migration of pTr cells, but these effects were blocked by SB203580, U0126, rapamycin, and LY294002, which inhibit p38 MAPK, ERK1/2, mTOR, and PI3K, respectively. These results suggest that VEGF is critical to successful growth and development of pTr during early pregnancy and that VEGF-induced stimulatory effect is coordinately regulated by multiple cell signaling pathways, including PI3K-AKT1 and MAPK signaling pathways.

Expression and regulation of avian cathepsin L in the oviduct during molting.

Cathepsins (CTSs) are peptidases that have biological roles in degrading extracellular matrix, catabolism of intracellular proteins, and processing of pro-hormones. Of these, cathepsin L (CTSL) is closely associated with morphological changes in reproductive organs required for proper function in mammals, including humans and mice, but little is known about CTSL in avian species. In the present study, the expression of CTSL was investigated in the oviduct of hens during regression and recrudescence in response to molting. Our results revealed that expression of CTSL mRNA increased (P<0.001) when the oviduct underwent regression during the molting period in hens. In situ hybridization and immunohistochemial analyses detected CTSL mRNA and protein predominantly in the luminal (LE) and glandular epithelia (GE) during regression of the oviduct, but not during regeneration of the oviduct. Expression of CTSL decreased in the oviduct of chicks treated with diethylstilbestrol (DES, a synthetic estrogen agonist). Furthermore, we discovered four miRNAs including miR-23b, miR-551, miR-1464 and miR-1803 that regulate expression of the CTSL gene at the post-transcriptional level, which suggests that CTSL mRNA can be regulated by specific miRNAs via 3'-UTR in chickens. Results of the present research suggest that estrogen regulates expression of CTSL during regression of the oviduct during molting and that down-regulation of CTSL is likely a prerequisite for the normal regeneration of oviductal tissues following molting in laying hens.

Sex-specific expression of CTNNB1 in the gonadal morphogenesis of the chicken.

BACKGROUND: Beta-catenin (CTNNB1), as a key transcriptional regulator in the WNT signal transduction cascade, plays a pivotal role in multiple biological functions such as embryonic development and homeostasis in adults. Although it has been suggested that CTNNB1 is required for gonad development and maintenance of ovarian function in mice, little is known about the expression and functional role of CTNNB1 in gonadal development and differentiation in the chicken reproductive system. METHODS: To examine sex-specific, cell-specific and temporal expression of CTNNB1 mRNA and protein during gonadal development to maturation of reproductive organs, we collected left and right gonads apart from mesonephric kidney of chicken embryos on embryonic day (E) 6, E9, E14, E18, as well as testes, oviduct and ovaries from 12-week-old and adult chickens and performed quantitative PCR, in situ hybridization, and immunohistochemical analyses. In addition, localization of Sertoli cell markers such as anti-Müllerian hormone (AMH), estrogen receptor alpha (ESR1), cyclin D1 (CCND1) and N-cadherin (CDH2) during testicular development was evaluated. RESULTS: Results of the present study showed that CTNNB1 mRNA and protein are expressed predominantly in the seminiferous cords on E6 to E14 in the male embryonic gonad, and are mainly localized to the medullary region of female embryonic gonads from E6 to E9. In addition, CTNNB1 mRNA and protein are abundant in the Sertoli cells in the testes and expressed predominantly in luminal epithelial cells of the oviduct, but not in the ovaries from 12-week-old and adult chickens. Concomitant with CTNNB1, AMH, ESR1, CCND1 and CDH2 were detected predominantly in the seminiferous cord of the medullary region of male gonads at E9 (after sex determination) and then maintained or decreased until hatching. Interestingly, AMH, ESR1, CCND1 and CDH2 were located in seminiferous tubules of the testes from 12-weeks-old chickens and ESR1, CCND1 and CDH2 were expressed predominantly in the Sertoli cells within seminiferous tubules of adult testes. CONCLUSIONS: Collectively, these results revealed that CTNNB1 is present in gonads of both sexes during embryonic development and it may play essential roles in differentiation of Sertoli cells during formation of seminiferous tubules during development of the testes.

Differential expression of secreted phosphoprotein 1 in response to estradiol-17ß and in ovarian tumors in chickens.

Secreted phosphoprotein 1 (SPP1), a highly phosphorylated protein containing a polyaspartic acid sequence and a conserved RGD motif, plays important roles in physiological processes such as inflammatory responses, calcification, organ development, immune cell function and carcinogenesis. Results of the present study indicate expression of SPP1 mRNA in various organs such as oviduct, small intestine and kidney from chickens, particularly in the glandular epithelium (GE) of the shell gland and, to a lesser extent, in luminal epithelium (LE) of the infundibulum and magnum, and GE of the isthmus of the oviduct. We determined that DES (diethylstilbestrol, a synthetic nonsteroidal estrogen) decreases SPP1 expression in the oviduct and that SPP1 mRNA and protein are significantly more abundant in GE of ovarian endometrioid carcinoma, but not the other cancerous and normal ovaries of hens. Further, microRNA-140 was discovered to influence SPP1 expression via its 3'-UTR which suggests that post-transcriptional regulation influences SPP1 expression in chickens. Collectively, results of this study indicate that SPP1 is novel in that its expression is down-regulated by estrogen in epithelial cells of the chicken oviduct and that it is up-regulated in chicken ovarian endometrioid tumor that could be used for monitoring effects of therapies for this disease in laying hens.

Differential expression of select members of the SLC family of genes and regulation of expression by microRNAs in the chicken oviduct.

The yolk and white of eggs from chickens contain proteins and other molecules either secreted or transported by cells of the reproductive tract, or secreted by the liver and transported to the ovarian follicles of laying hens. Nutrients transported by solute carriers (SLCs) include glucose, electrolytes, and amino acids. Although SLC genes have been investigated in mammals, there are few studies of expression of SLC genes in the chicken oviduct. Therefore, we investigated temporal and cell-specific expression of selected SLC genes at 3 h and 20 h postovulation and regulation of their expression by microRNAs (miRs). Expression of SLC1A4 (glutamate and neutral amino acid transporter), SLC13A2 (dicarboxylate transporter), and SLC35B4 (UDP-xylose: UDP-N-acetylglucosamine transporter) mRNAs was limited to glandular epithelium (GE), while SLC4A5 (sodium bicarbonate cotransporter) and SLC7A3 (cationic amino acid transporter) mRNAs were expressed predominantly in the luminal epithelium of the magnum. Interestingly, SLC1A4, SLC4A5, SLC13A2 and SLC35B4 mRNAs were abundant only in GE of the shell gland, whereas SLC7A3 was not detected in the shell gland. In the magnum, SLC7A3 and SLC4A5 were expressed, but SLC1A4, SLC35B4, and SLC13A2 were not expressed at 20 h postovulation. In the shell gland, all SLC mRNAs were expressed at both time points, except for SLC7A3. The miRNA target validation assay revealed that miR-1764 and miR-1700 bind directly to SLC13A2 and SLC35B4 transcripts, respectively, to regulate expression. Results of this study demonstrate cell-specific and temporal changes in expression of selected SLC genes and regulation of SLC13A2 and SLC35B4 expression by miRs in the oviduct of laying hens.

Cell-specific and temporal aspects of gene expression in the chicken oviduct at different stages of the laying cycle.

Egg formation and embryonic development occur as the yolk passes through the magnum, isthmus, and shell gland of the oviduct before oviposition in hens. The present study identified candidate genes associated with secretory function of the chicken oviduct after ovulation and contributing to egg formation and oviposition. Hens (n = 5 per time point) were euthanized to recover the reproductive tract when the egg was in the magnum (3 h after ovulation) and the shell gland (20 h after ovulation). Total RNA was extracted from each segment of the oviducts and subjected to Affymetrix chicken GeneChip analysis. Quantitative PCR and in situ hybridization analyses of selected genes confirmed the validity of the gene expression patterns detected using microarray analysis. In particular, ACP1, CALB1, CYP26A1, PENK, RCAN1 and SPP1 expression increased significantly in the shell gland between 3 h and 20 h postovulation, whereas only RCNA1 expression increased significantly in the magnum between 3 h and 20 h postovulation. Results of the high-throughput analysis revealed cell-specific and temporal changes in gene expression in the oviduct at 3 h and 20 h postovulation in laying hens provide novel insight into changes at the molecular and cellular levels of candidate genes related to formation of the egg and oviposition.

ERBB receptor feedback inhibitor 1: identification and regulation by estrogen in chickens.

The ERBB receptor feedback inhibitor 1 (ERRFI1) is a scaffolding adaptor protein, that plays a pivotal role in the epidermal growth factor receptor (EGFR) cell signaling cascade as a negative regulator affecting many important physiological processes. It was recently reported that ERRFI1 is a critical regulator of the response of the endometrium to estrogen regulation of tissue homeostasis in mice. But, very little is known about ERRF11 and hormonal regulation of the ERRFI1 gene in chickens. Therefore, in the present study, ERRFI1 gene was cloned and its differential expression profile analyzed at different embryonic stages, in various adult organs, and in oviducts from estrogen-treated chickens. Chicken ERRFI1 has an open-reading frame of 2848 nucleotides that encode for a protein of 465 amino acids that has considerable homology to mammalian ERRFI1 proteins (>62% identity). Importantly, ERRFI1 mRNA is abundantly distributed in various organs from chickens. We then determined that DES (diethylstilbestrol, a synthetic nonsteroidal estrogen) induced ERRFI1 mRNA and protein predominantly in luminal and glandular epithelial cells of the oviduct. Further, we determined whether microRNAs, specifically miR-200b, miR-429 and miR-1639, influence ERRFI1 expression via its 3'UTR and found that it does not directly target the 3'UTR of ERRFI1 mRNA. Therefore, it is unlikely that post-transcriptional regulation influences ERRFI1 expression in the chicken oviduct. In conclusion, our results indicate that ERRFI1 is a novel estrogen-stimulated gene expressed in epithelial cells of the chicken oviduct that likely plays an important role in oviduct growth and differentiation during early development of the chicken.

Tissue specific expression and estrogen regulation of SERPINB3 in the chicken oviduct.

Serine protease inhibitors (SERPINs) comprise the largest superfamily of protease inhibitors and appear to be ubiquitously expressed in a variety of species. Of these, squamous cell carcinoma antigen 1 (SCCA1), also known as a SERPINB3, was first identified in squamous cell carcinoma tissue from the cervix of women. However, there is little known about the expression and hormonal regulation of SERPINB3 in chickens. Therefore, the avian SERPINB3 gene was compared with those of other species with respect to structure, phylogenetic evolution and tissue- and cell-specific expression in hens. Chicken SERPINB3 has moderate homology to mammalian SERPINB3 proteins (36-47%). Of particular note, SERPINB3 mRNA was most abundant in the chicken oviduct and cell-specific expression was in glandular (GE) and luminal (LE) epithelial cells of the oviduct of laying hens. Treatment of young chicks with DES (diethylstilbestrol, a synthetic nonsteroidal estrogen) induced SERPINB3 mRNA and protein in GE and LE, but not in other cell types of the oviduct. Western blot analyses determined that immunoreactive SERPINB3 protein was also increased by DES in LE and GE of the oviduct of chicks. Collectively, these results indicate that SERPINB3 is an estrogen-induced gene expressed only in LE and GE of the chicken oviduct and implicate SERPINB3 in regulation of oviduct development and egg formation.

Development of a fuzzy logic-controlled system for home cultivation of sweet basil.

As environmental pollution and the global population increase, and the COVID-19 pandemic becomes more severe, demands for indoor farming, especially home food gardening, have also increased. However, most research thus far has focused on large-scale food production, with very few studies having been conducted at the household scale. Also, the devices cultivating household crops with control systems in a continuous way, which minimize fluctuations of environmental conditions, have been rarely developed. Therefore, this study aimed to design a household cultivation system for sweet basil that is automatically and continuously controlled by fuzzy logic with a Raspberry Pi4. Three inputs (temperature, humidity, and growth stage) and seven outputs (fan, humidifier, heater 1, heater 2, LED red, green, and blue) were used with six rules, ensuring that three lights operated independently upon three growth stages. Simulation and actual operation were carried out, resulting in an appropriately controlled system that operated with few defects. In the case of an operation of the input variable, temperature and humidity were maintained at an average of 21.24 °C and 75.58%, respectively, and the LED operation for the growth stage was confirmed to be flawless. For verification of the designed fuzzy system, a comparison between the simulation and actual operation was performed to examine differences and identify problems. To this end, Pearson's correlation coefficients were used, and the direction of correction of the fuzzy logic system was proposed. Through these results, the feasibility of a home cultivation system using fuzzy logic was demonstrated, and it is expected that further studies applying it will be conducted in the future.

Hesperidin Suppresses the Proliferation of Prostate Cancer Cells by Inducing Oxidative Stress and Disrupting Ca2+ Homeostasis.

Although androgen deprivation therapy is mainly used for its treatment, the mortality rate of prostate cancer remains high due to drug resistance. Hence, there is a need to discover new compounds that exhibit therapeutic effects against prostate cancer with minimum side effects. Hesperidin is a flavonoid carbohydrate isolated from citrus fruits. It has antiproliferative effects in various cancer types; however, whether it can modulate cell proliferation by modulating the key targets of cancer therapy, including intracellular signaling pathways and oxidative stress, remains unknown. Therefore, we confirmed that hesperidin suppressed the proliferation of prostate cancer cells, PC3 and DU145. Hesperidin induced cell death by regulating the cell cycle and inhibited the expression of proliferating cell nuclear antigen, a cell proliferation marker. Hesperidin also promoted the generation of reactive oxygen species and induced mitochondrial membrane depolarization and endoplasmic reticulum stress in prostate cancer cells. Moreover, as hesperidin increased Ca2+ levels in prostate cancer cells, we co-treated the inositol 1,4,5-trisphosphate receptor inhibitor, 2-aminoethyl diphenyl borate (2-APB), with hesperidin. Notably, 2-APB restored cell proliferation, which was reduced to control levels by hesperidin. In addition, hesperidin inhibited the activation of the phosphoinositide 3-kinase and mitogen-activated protein kinase signaling pathways. Hesperidin also enhanced the anticancer effects of the chemotherapeutic agent, cisplatin, in both PC3 and DU145 cells. Taken together, these results suggest that hesperidin can be used as a potential therapeutic adjuvant in prostate cancer as it can inhibit cell proliferation by mediating oxidative stress and increasing Ca2+ levels.

Avian SERPINB11 gene: characteristics, tissue-specific expression, and regulation of expression by estrogen.

Serpins, a group of proteins with similar structural and functional properties, were first identified based on their unique mechanism of action: their inhibition of proteases. While most serpins have inhibitory roles, certain serpins are not involved in canonical proteolytic cascades but perform diverse functions including storage of ovalbumin in egg white, transport of hormones (thyroxine- and cortisol-binding globulin), and suppression of tumors. Of these, serpin peptidase inhibitor, clade B, member 11 (SERPINB11) is not an inhibitor of known proteases in humans and mice, and its function is unknown. In the present study, the SERPINB11 gene was cloned, and its expression profile was analyzed in various tissues from chickens. The chicken SERPINB11 gene has an open reading frame of 1346 nucleotides that encode a protein of 388 amino acids that has moderate homology (38.8%-42.3%) to mammalian SERPINB11 proteins. Importantly, SERPINB11 mRNA is most abundant in the chicken oviduct, specifically luminal and glandular epithelia, but it was not detected in any other chicken tissues of either sex. We then determined effects of diethylstilbestrol (DES; a synthetic nonsteroidal estrogen) on SERPINB11 expression in the chicken oviduct. Treatment of young chicks with DES induced SERPINB11 mRNA and protein only in luminal and glandular epithelial cells of the oviduct. Collectively, these results indicate that the novel estrogen-induced SERPINB11 gene is expressed only in epithelial cells of the chicken oviduct and implicate SERPINB11 in regulation of oviduct development and differentiated functions.