Embryo Liver Research Articles

Developmental toxicity and estrogenic activity of antimicrobial phenolic-branched fatty acids using in silico simulations and in vivo and in vitro bioassay.

Due to the growing safety and environmental concerns associated with biocides, phenolic-soy branched chain fatty acids (phenolic-soy BCFAs) are synthesized as new bio-based antimicrobial agents. Safety evaluation is essential before the wide adoption of these new antimicrobial products. This study was initiated to evaluate the safety of four phenolic-soy BCFAs (with phenol, thymol, carvacrol, or creosote branches). Methyl-branched iso-oleic acid, phenol, and creosote were included in the study as controls. In silico toxicity simulation tools predicted that the phenolic BCFAs had much higher toxicities to aquatic organisms than free phenolics did, while the opposite was predicted for rats. The developmental toxicity of four phenolic-soy BCFAs was assessed using an in vivo chicken embryonic assay. Results showed that creosote-soy BCFA had much lower mortality rates than creosote at the same dosages. Additionally, creosote-soy BCFA and methyl-branched iso-oleic acid induced minimal estrogenic activity in the concentration range of 10nM - 1µM. Carvacrol-soy BCFA treatments significantly increased (p < 0.05) oxidative stress levels with higher thiobarbituric acid reactive substances in the livers of chicken embryos. Altogether, the phenolic-soy BCFAs, especially creosote-soy BCFA, reported in this study are potentially promising and safer bio-based antimicrobial products.

Nutritional supplementation of breeding hens may promote embryonic development through the growth hormone-insulin like growth factor axis

The late stage of embryo development is a crucial period of metabolic changes, with rapid organ development requiring a substantial supply of nutrients. During this phase, maternal nutritional levels play a vital role in the growth, development, and metabolism of the offspring. In this study, we added 2 doses of β-carotene (βc) (120 mg/kg and 240 mg/kg) to the daily diet of Hailan Brown laying hens to investigate the impact of maternal nutritional enrichment on embryo development. Maternal nutrition supplementation significantly increased the expression of chicken embryo liver index, growth hormone (GH), insulin-like growth factor-1 (IGF-1), and hepatocyte growth factor (HGF) in serum. At the same time, the expression of GH/growth hormone receptor (GHR), IGF-1 mRNA, and Proliferating Cell Nuclear Antigen (PCNA) protein in the liver was upregulated, indicating that maternal nutrition intervention may promote chicken embryo liver development through the GH-IGF-1 axis. Transcriptome sequencing results showed that differential genes in liver after maternal nutritional supplementation with β-carotene were enriched in pathways related to cell proliferation and metabolism. Consequently, we postulated that maternal β-carotene supplementation might operate via the GH-IGF-1 axis to regulate the expression of genes involved in growth and development, thereby promoting liver development. These results contribute to formulating more effective poultry feeding strategies to promote offspring growth and development.

Attenuation and molecular characterization of fowl adenovirus 8b propagated in a bioreactor and its immunogenicity, efficacy, and virus shedding in broiler chickens

Background and Aim: Live-attenuated vaccines are the most successful type of vaccine and could be useful in controlling fowl adenovirus (FAdV) 8b infection. This study aimed to attenuate, molecularly characterize, and determine the immunogenicity, efficacy, and challenge virus shedding in broiler chickens. Materials and Methods: The FAdV 8b isolate (UPM08136) was passaged onto chicken embryo liver (CEL) cells until attenuation. We sequenced and analyzed the hexon and fiber genes of the passage isolates. The attenuated bioreactor-passage isolate was inoculated into 1-day-old broiler chickens with (attenuated and inactivated) and without booster groups and challenged. Body weight (BW), liver weight (LW), liver: body weight ratio (LBR), FAdV antibody titers, T-lymphocyte subpopulation in the liver, spleen, and thymus, and challenge virus load and shedding were measured. Results: Typical cytopathic effects with novel genetic changes on CEL cells were observed. The uninoculated control-challenged (UCC) group had significantly lower BW and higher LW and LBR than the inoculated groups. A significantly higher FAdV antibody titer was observed in the challenged non-booster and attenuated booster groups than in the UCC group. T cells in the spleen and thymus of the liver of inoculated chickens were higher than uninoculated control group levels at all-time points and at different times. A significantly higher FAdV challenge virus load was observed in the liver and shedding in the cloaca of UCC chickens than in non-booster chickens. Conclusion: The FAdV 8b isolate was successfully attenuated, safe, and immunogenic. It reduces virus shedding and is effective and recommended as a vaccine against FAdV infection in broiler chickens. Keywords: antibodies, attenuation, fowl adenovirus 8b, T lymphocytes, vaccines, virus shedding.

Liver and kidney damage in quail embryos hatched from eggs sprayed once with chlorpyrifos during artificial incubation.

Birds have enough conditions to be used as bioindicators for the presence of environmental contaminants. Notably, chlorpyrifos (CPF) remains extensively employed in Argentina, particularly in fruit plantations and livestock settings This study aimed to assess the potential impacts of CPF on common quail (Coturnix coturnix) embryos through external eggshell application during key embryonic stages (days 1, 4, and 14). Concentrations equivalent to those used in fruit applications, 5× and 10 × (38.4, 192, and 384µg/egg), were employed. A 48% commercial formulation of CPF (Pirfos - Icona®) was utilized. An increase in embryonic deaths, as well as a statistical association between the degree of liver or kidney lesions and CPF concentrations was observed. The results suggest that CPF can induce embryotoxic effects with a single exposure to quail embryos and compromise the viability of the organisms. The study suggests a potential exposure risk for embryos through contact with the insecticide on the eggshell's exterior.

Hepatic effects of acetochlor chiral isomers in zebrafish and L02 cells

The pesticide acetochlor (ACT) is a chiral isomer commonly detected in the global environment, yet its specific impacts on liver function remain poorly understood. We utilized zebrafish and L02 cells as research models to comprehensively investigate how ACT and its chiral isomers affect the liver. Our investigations unveiled that the R, Rac, and S isomers of ACT disrupt hepatic lipid transport, catabolism, and synthesis, leading to delayed yolk sac absorption and the accumulation of lipids in zebrafish embryos. These isomers induce oxidative stress in the liver of zebrafish embryos, reducing antioxidant levels and enzyme activity. The accumulated lipids in the liver render it susceptible to oxidative stress, further exacerbating hepatocyte damage. Hepatocyte damage manifests as extensive vacuolization of liver cells and alterations in liver morphology, which are induced by R, Rac, and S. Furthermore, we elucidated the molecular mechanisms underpinning the disturbance of hepatic lipid metabolism by R, Rac, and S in L02 cells. These compounds stimulate lipid synthesis through the upregulation of the AMPK/SREBP-1c/FAS pathway while inhibiting lipolysis via downregulation of the PPAR-α/CPT-1a pathway. Remarkably, our results highlight that S exhibits significantly higher hepatotoxicity in comparison to R. This study provides valuable insights into the hepatic effects of ACT chiral isomers.

Unveiling the Cell Wall-Targeting Mechanisms and Multifaceted Virulence Modulation by a Eugenol Glycoconjugate against Aspergillus fumigatus: Insights from in vitro and in ovo Studies.

The primary objective of this study was to elucidate the putative cell wall-associated targets of compound 6i, a glycoconjugate of eugenol, in Aspergillus fumigatus, while also evaluating its toxicity and assessing histopathologic alterations in the liver, heart, and kidney of compound 6i-treated embryos using an in ovo model. To achieve this aim, compound 6i was synthesized, and a series of biochemical assays were performed to determine its impact on the fungal cell wall. Additionally, qRT-PCR and LC-MS/MS analyses were conducted to investigate changes in gene and protein expression profiles associated with melanin biosynthesis, conidiation, siderophore production, transcriptional regulation of β-glucan biosynthesis, and calcineurin activity in A. fumigatus. The experimental findings revealed that compound 6i exhibited notable antifungal activity against A. fumigatus by perturbing cell wall integrity, hindering ergosterol, glucan, and chitin biosynthesis, and inhibiting catalase production. Moreover, relative gene expression and proteomic analyses demonstrated that compound 6i exerted both down-regulatory and up-regulatory effects on several crucial genes and proteins involved in the aforementioned fungal processes. Furthermore, increased expression of oxidative stress-related proteins was observed in the presence of compound 6i. Notably, the glycoconjugate of eugenol did not elicit cytotoxicity in the liver, heart, and kidney of chick embryos. The current investigation elucidated the multifaceted mechanisms by which compound 6i exerts its antifungal effects against A. fumigatus, primarily through targeting cell wall components and signaling pathways. These findings underscore the potential of the eugenol glycoconjugate as a promising antifungal candidate, warranting further exploration and development for combating A. fumigatus infections.

Efficacy, immunogenicity, and virus shedding in broiler chickens inoculated with live attenuated fowl adenovirus serotype 8b propagated a bioreactor.

Fowl adenovirus (FAdV) 8b causes huge economic losses in the poultry industry worldwide. Attenuated FAdV 8b could be useful in preventing FAdV infections globally and scale-up obstacles could be solved by bioreactor technology. This study was carried out to attenuate the FAdV 8b isolate, propagate it in a bioreactor, molecularly characterize the passage isolates, and determine the immunogenicity, efficacy, and shedding of the virus of chickens. FAdV serotype 8b (UPM11142) isolate was passaged on chicken embryo liver (CEL) cells until attenuation and propagated in a bioreactor (UPM11142P20B1). Hexon and fiber genes of the isolates were sequenced and analyzed. UPM11142P20B1 was administered to 116-day-old broiler chickens divided into four groups, A (control), B (non-booster), C (booster with UPM11142P20B1), and D (booster with inactivated UPM11142P5B1). Eight chickens from each group were challenged. Body weight (BW) and liver weight (LW), liver: BW ratio (LBR), FAdV antibody titer, T lymphocyte sub-populations in the liver, spleen and thymus; and challenge virus load in the liver and shedding in cloaca were measured at weekly intervals. The isolate caused typical cytopathic effects on CEL cells typical of FAdV. Novel molecular changes in the genes occurred which could be markers for FAdV 8b attenuation. BW, LW, and LBR were similar among groups throughout the trial but the uninoculated control-challenged group (UCC) had significantly higher LBR than the inoculated and challenged groups at 35 dpi. Non-booster group had higher FAdV antibodies at all time points than the uninoculated control group (UCG); and the challenged booster groups had higher titer at 35 dpi than UCC. T lymphocytes increased at different time-points in the liver of inoculated chickens, and in the spleen and thymus as well, and was higher in the organs of inoculated challenged groups than the UCC. There was a significantly higher challenge virus load in the liver and cloaca of UCC chickens than in the non-booster chickens. UPM11142P20B1 was safe, efficacious, significantly reduced shedding, and is recommended as a candidate vaccine in the prevention and control of FAdV 8b infections in broiler chickens.

The Importance and Challenges of Primary Chicken Embryo Liver Cells in Studies of Poultry Viral Diseases: A Review

Primary chicken embryo liver (CEL) cells are derived from the liver tissue of chicken embryonated eggs (CEE) using an aseptic isolation technique and growth under a controlled atmosphere in an artificial environment for cell attachment and proliferation. Although this primary cultured cell has been established for more than six decades, utilization of primary cells is still the preferable medium nowadays as the “gold standard” due to several advantages over other diagnostic techniques. Cells provide better adaptability of the viruses and easily mimic the natural host environment with high virus titration. The volume of virus suspension could be increased by applying an immortal chicken embryo liver-derived cell line. The current review aimed to highlight the importance and challenges of using primary chicken embryo liver cells in poultry virus studies. Primary CEL cells are widely used as an alternative host for diagnosis of infectious poultry viruses, cultivation and passaging of virus isolates, and vaccine production. Yet, there are some challenges and limitations in handling this primary cell, which requires appropriate facilities and environment to sustain the rapid growth of confluent monolayer cells, as highlighted in this paper. The availability of specific pathogen-free CEE is a major concern due to limited resources globally, thus creating a challenge for vaccine manufacturers to upscale the cultured cells. Future improvement of primary cell culture preparation necessitates new technology by applying cellular microcarrier in the bioreactor machine for efficient cell growth and subsequent routine virus cultivation. This study can help the researchers understand the advantages of primary CEL cells and their applications due to their significant impact on poultry viruses.

Attenuation of a Field Strain of Infectious Laryngotracheitis Virus in Primary Chicken Culture Cells and Adaptation to Secondary Chicken Embryo Fibroblasts

The establishment of commercial infectious laryngotracheitis virus (ILTV) live-modified vaccines has relied on serial passaging in chicken embryo (CEO) and tissue culture (TCO) for attenuation. The objective of this study was to attenuate and adapt a virulent CEO-related ILTV field strain (6340) in immortalized cells (LMH), primary chicken embryo kidney cells (CEK), chicken embryo liver cells (CEL), and chicken embryo fibroblasts (CEF). CEFs were refractory to parent ILTV, LMH cells produced low virus yields (~2.5 log10 TCID50 per mL), while CEK and CEL cells produced higher viral titers (≥log10 6.0 TCID50 per mL). After 52 passages in CELs, the cytopathic effect (CPE) was observed not only in hepatocytes but also in CEL fibroblasts. Once CPE was evident in CEL fibroblasts, 20 further passages in CEFs with viral titers reaching yields of ~4.4–5.5 log10 TCID50 per mL were performed. The attenuation of CEF-adapted viruses was evaluated after intra-tracheal and conjunctival inoculation in 28-day-old broilers by assessing clinical signs at five days post-inoculation (DPI). Virus CEL cells passages 80, 90, and 100, and CEF passages 10 and 20 were significantly attenuated compared to the parental strain. This is the first report of the attenuation of a virulent field CEO-related ILTV strain (RFLP Group V) in CEF cells—a cell type from a different embryonic germ layer (mesoderm) than ILTV target cells—the respiratory epithelium (endoderm). This finding underscores the potential use of CEF adaptation for the development of a live-attenuated ILTV vaccine.

CK2β Regulates Hematopoietic Stem Cell Biology and Erythropoiesis.

The Ser-Thr kinase CK2 plays important roles in sustaining cell survival and resistance to stress and these functions are exploited by different types of blood tumors. Yet, the physiological involvement of CK2 in normal blood cell development is poorly known. Here, we discovered that the β regulatory subunit of CK2 is critical for normal hematopoiesis in the mouse. Fetal livers of conditional CK2β knockout embryos showed increased numbers of hematopoietic stem cells associated to a higher proliferation rate compared to control animals. Both hematopoietic stem and progenitor cells (HSPCs) displayed alterations in the expression of transcription factors involved in cell quiescence, self-renewal, and lineage commitment. HSPCs lacking CK2β were functionally impaired in supporting both in vitro and in vivo hematopoiesis as demonstrated by transplantation assays. Furthermore, KO mice developed anemia due to a reduced number of mature erythroid cells. This compartment was characterized by dysplasia, proliferative defects at early precursor stage, and apoptosis at late-stage erythroblasts. Erythroid cells exhibited a marked compromise of signaling cascades downstream of the cKit and erythropoietin receptor, with a defective activation of ERK/JNK, JAK/STAT5, and PI3K/AKT pathways and perturbations of several transcriptional programs as demonstrated by RNA-Seq analysis. Moreover, we unraveled an unforeseen molecular mechanism whereby CK2 sustains GATA1 stability and transcriptional proficiency. Thus, our work demonstrates new and crucial functions of CK2 in HSPC biology and in erythropoiesis.

Epidemiology and zoonotic potential of Livestock-associated Staphylococcus aureus isolated at Tamil Nadu, India

BackgroundStaphylococcus aureus is part of normal flora and also an opportunistic pathogen responsible for a wide range of infections in both humans and animals. Livestock-associated S. aureus (LA-SA) has gained importance in recent years due to its increased prevalence in recent years, becoming a worry in public health view. This study aimed to study the epidemiology of LA-SA strains in Madurai district, Tamil Nadu, India.MethodsA total of 255 samples were collected from bovine and other small ruminants like goats and sheep nares (n = 129 and n = 126 respectively). Nasal swab samples were collected from study animals with sterile sample collecting cotton swabs (Hi-Media, Mumbai). Samples were transported to the lab in Cary-Blair Transport media for further analysis. The samples were tested for S. aureus using antibiotic selection and PCR-based assays. The pathogenicity of the bacteria was assessed using chicken embryo models and liver cross-sections were used for histopathology studies.ResultsThe prevalence rate in bovine-associated samples was 42.63% but relatively low in the case of small ruminants associated samples with 28.57% only. The overall prevalence of S. aureus is found to 35.6% and MRSA 10.98% among the study samples. The antibiogram results that LA-SA isolates were susceptible to aminoglycosides and tetracyclines but resistant to β-lactam drugs. The biofilm formation results showed that the LA-SA isolates are weak to high-capacity biofilm formers. The enterotoxigenic patterns revealed that most of the isolated strains are enterotoxigenic and possess classical enterotoxins. The survival analysis of chicken embryos suggested that the Bovine-associated strains were moderately pathogenic.ConclusionThe study concluded that economically important livestock animals can act as reservoirs for multi-drug resistant and pathogenic which in-turn is a concern for public health as well as livestock health.

Estrogens and Prolactin Do Not Regulate Maternal and Embryo Iron Homeostasis during Pregnancy

Iron is essential for maternal and fetal health but the molecular mechanisms ensuring increased iron availability during pregnancy are not well understood. Hepcidin is the key iron-regulatory hormone and functions by blocking iron absorption, recycling and mobilization from stores, effectively decreasing plasma iron levels. In healthy human and rodent pregnancies, maternal hepcidin decreases starting in the second trimester and is nearly undetectable by late pregnancy. This decrease in hepcidin allows for greater iron availability for transfer to the fetus. If hepcidin is not appropriately suppressed during pregnancy, it results in fetal anemia, intrauterine growth restriction, and even fetal demise in severe cases 1. Unrelated to pregnancy, hormones estrogen and prolactin have been previously implicated in hepcidin regulation 2,3, although with conflicting results on the direction of hepcidin change 4-6. The levels of both hormones increase dramatically over the course of pregnancy, thus we hypothesized that they may modulate iron homeostasis during pregnancy. Studying the role of estrogens and prolactin during pregnancy is challenging as both hormones are required for pregnancy. Additionally, mice with global knockout (KO) of either estrogen receptor alpha or prolactin receptor are infertile. Thus, to determine the contribution of estrogens and prolactin on maternal hepcidin suppression during pregnancy, we generated liver-specific knockouts of either estrogen receptor alpha ( Er- α f/f;Alb-Cre +) or prolactin receptor ( Prlr f/f;Alb-Cre +), here abbreviated as ER-α KO and PRLR KO. Female ER-α KO, PRLR KO and WT mice were set up for timed pregnancy, and liver hepcidin mRNA, as well as maternal and embryo iron and hematological parameters were measured at E18.5 . Hepcidin suppression in ER-α KO and PRLR KO dams was not different from that in WT dams, suggesting that estrogens and prolactin are not major mediators of maternal hepcidin suppression. Additionally, maternal and embryo hematological parameters and liver iron concentrations were similar between WT, ER-α KO, PRLR KO pregnancies, further supporting our observation that estrogens and prolactin do not regulate iron homeostasis in pregnancy. The mechanisms by which maternal hepcidin is suppressed during pregnancy remain to be determined. 1Sangkhae V et al, Blood, 2020 2Yang Q et al, Endocrinology, 2012 3Wang J et al, Nanomedicine: nanotechnology, biology and medicine, 2015 4Zhen et al, Hepatology, 2013 5Ikeda Y et al, PloS one, 2012 6Lehtihet M et al, PloS one, 2016

Effects of silver nanoparticles prepared by aqueous extract of Ferula communis on the developing mouse embryo after maternal exposure.

Green synthesis of silver nanoparticles (AgNPs) from aqueous silver nitrate has been achieved using an extract of Ferula communis leaf as a capping, reducing, and stabilizing agent. The formation and stability of the green synthesized silver nanoparticles in the colloidal solution were monitored by absorption measurements. Silver nanoparticles were characterized by different analyses such as X-ray diffraction (XRD), energy dispersive spectroscopy (EDS), and FT-IR spectroscopy. The average particle size of silver nanoparticles was determined by high-resolution transmission electron microscopy (HRTEM) and scanning electron microscopy (SEM) analyses. In this experiment, pregnant female mice were divided into four groups (G); G1 was the control and received phosphate-buffered saline, G2 received orally aqueous extract of F. communis leaf, G3 received orally AgNPs chemically prepared by NaBH4, and G4 received orally AgNPs prepared by aqueous extract of F. communis leaf. The diameter of AgNPs was 20nm. AgNPs exhibited good catalytic reduction ability toward methyl orange in the presence of sodium borohydride with a rate constant of 2.95 x 10-4 s-1. The results revealed the occurrence of resorbed embryos in G2, G3, and G4 with different percentages. The livers of mothers and embryos at E14.5 in G2, G3, and G4 showed different levels of histopathological alteration and increase in GFAP and CTGF expressions compared with the control group. The study concluded that the oral administration of small-sized AgNPs (20nm) prepared by Ferula extract had less toxicity than those prepared by the chemical method.

Curcumin-functionalized gold nanoparticles attenuate AAPH-induced acute cardiotoxicity via reduction of lipid peroxidation and modulation of antioxidant parameters in a chicken embryo model

Gold nanoparticles (AuNPs) have gained considerable attention due to their biocompatibility, customizable optical properties and ease of synthesis. In this study, an environmentally friendly method was used for synthesize curcumin-functionalized AuNPs (AuNP-C). AuNP-C exhibited a spherical shape, uniformity, and an average diameter of 6 nm. The in vitro antioxidant activity was analyzed, and cytotoxicity properties of AuNP-C were assessed in fibroblast and macrophage cells. Additionally, the effects of AuNP-C on oxidative stress in chicken embryo liver and hearts were investigated. AuNP-C demonstrated potent free radical scavenging properties without exhibiting cytotoxicity and hepatotoxicity effects. Administration of 300 µg/mL of AuNP-C in chicken embryos, subjected to oxidative damage induced by 2,2′-azobis(2-amidinopropane) dihydrochloride, significantly reduced lipid peroxidation and reactive oxygen species levels in the cardiac tissue. Moreover, the activities of cardiac superoxide dismutase, catalase, and glutathione reductase were restored, accompanied by an increase in overall antioxidant capacity. Furthermore, at higher concentrations, AuNP-C normalized the reduced glutathione content. AuNP-C preserved the normal structure of blood vessels; however, it resulted in an increase in protein carbonylation. This study provides initial evidence for the modulation of antioxidant defense mechanisms by green-synthesized AuNPs and underscores the importance of investigating the in vivo safety of phytoantioxidant-functionalized nanoparticles.

Remimazolam relieved the injury of hypoxia/reperfusion treated human embryo liver cell line through the targeting METTL3 mediated m6A modification of P53

BackgroundThis study was performed to explore the role of Re in liver IRI progression. Hypoxia and reperfusion (H/R) treated human embryo liver cell line (L-02) was used to establish a liver IRI model. Materials and methodsCell behaviors were detected using CCK-8, flow cytometry and TUNEL staining assays. The m6A content was detected using m6A dot blot assay. RT-qPCR and western blot assays were used to assessed the relative mRNA and protein levels. MeRIP assay was conducted to determine the m6A levels of P53. The relationship between METTL3 and P53 was demonstrated using RIP and dual-luciferase reporter assays. ResultsThe results showed that Re treatment significantly decreased the cell apoptosis and promoted the cell viability in the H/R treated L-02 cells. Besides, H/R treatment increased the METTL3 and m6A levels in the L-02 cells, and Re treatment decreased them. Additionally, METTL3 overexpression reversed the role of Re in the H/R treated L-02 cells. Mechanistically, METTL3 overexpression enhanced the m6A levels and mRNA stability and expressions of P53. The combination of METTL3 and P53 was further confirmed. ConclusionIn conclusion, this study demonstrated that Re treatment relieved the H/R induced injury in the L-02 cells through decreasing the METTL3 levels. METTL3 enhanced the mRNA stability and expressions of P53 through m6A modification. Re-METTL3-P53 axis might a new direction for the treatment of liver IRI in the future.

Vitamin E and A Availability in Goose Embryos and Goslings and Improvement of Reproduction Traits Depending on the Starting Temperature Regime of Egg Incubation

One of the major problems impeding the sustainable development of goose production is low egg hatchability. Thus, it is imperative to develop more efficient ways to improve the hatching qualities of goose eggs. The present study aimed to investigate the effects of changes in the temperature regime of incubation on the availability of vitamins A (retinol) and E (α-tocopherol), as well as important elements of antioxidant protection in embryos and goslings. The initial heating of eggs at temperatures of 39 and 39.5 °C during the first 24 or 36 h was used as the main factor influencing goose embryo development. The dynamics in the content of antioxidant retinol and α-tocopherol were analyzed in the liver of embryos and goslings (up to 9 days of age) using the standard regime of incubation (at 38 °C) and the prior heating of eggs (at 39 °C and 39.5 °C) in the first 24 and 36 h of incubation. The obtained results provided new information about the effect of the initial heating of eggs on the function of antioxidants in the gosling’s body. It was confirmed that the age-related changes of retinol and α-tocopherol levels in goslings are similar to those found in chicks, ducklings, and turkey-poults. In addition, the effect of an egg’s weight on hatchability was established, with the hatchability of eggs in the medium weight class being higher than that of smaller and larger eggs. The efficiency of the redistribution of retinol from the yolk to the liver in day-old goslings was higher by 6% when exposing eggs to a temperature of 39.5 °C for 36 h. The initial heating regime at 39.5 °C led to an increased hatch of goslings, and to the evenness of their hatch from eggs of different weights, approaching the hatch values of the medium class. Overall, the α-tocopherol concentration in the liver of embryos and goslings, using the starting heating of eggs, was higher than that with the standard temperature mode of incubation. At the same time, the hatch rate of goslings increased by 9–13%, and the hatchability of eggs by 10–16%. For use in the practice of hatcheries and breeding poultry farms, it is recommended that goose eggs are heated for 36 h at 39.5 °C.

ISG15 suppresses ovulation and female fertility by ISGylating ADAMTS1

BackgroundISGylation is a post-translational protein modification that regulates many life activities, including immunomodulation, antiviral responses, and embryo implantation. The exact contribution of ISGylation to folliculogenesis remains largely undefined.ResultsHere, Isg15 knockout in mice causes hyperfertility along with sensitive ovarian responses to gonadotropin, such as increases in cumulus expansion and ovulation rate. Moreover, ISG15 represses the expression of ovulation-related genes in an ISGylation-dependent manner. Mechanistically, ISG15 binds to ADAMTS1 via the ISG15-conjugating system (UBA7, UBE2L6, and HERC6), ISGylating ADAMTS1 at the binding sites Lys309, Lys593, Lys597, and Lys602, resulting in ADAMTS1 degradation via a 20S proteasome-dependent pathway.ConclusionTaken together, the present study demonstrates that covalent ISG15 conjugation produces a novel regulatory axis of ISG15-ADAMTS1 that enhances the degradation of ADAMTS1, thereby compromising ovulation and female fertility.

Lung fusion with the liver in chick and quail embryos.

Even in mammals with the diaphragm, the lung and liver are likely to attach mutually without separation by any structure in embryos. The aim of this study was to examine whether or not the lung attaches to the liver in embryonic development of birds without diaphragm. First, we ensured the topographical relation between the lung and liver in 12 human embryos at 5 weeks. After the serosal mesothelium was established, the human lung sometimes (3 embryos) attached tightly to the liver without interruption by the developing diaphragm in the pleuroperitoneal fold. Second, we observed the lung-liver interface in chick and quail embryos. At 3-5 days' incubation (stages 20-27), the lung and liver were fused at bilateral narrow areas just above the muscular stomach. Therein, mesenchymal cells, possibly derived from the transverse septum, were intermingled between the lung and liver. The interface tended to be larger in the quail than the chick. At and until 7 days' incubation, the fusion of the lung and liver disappeared and, instead, a membrane connected them bilaterally. The right membrane extended caudally to attach to the mesonephros and caudal vena cava. At 12 days' incubation, bilateral thick folds, containing the abdominal air sac and pleuroperitoneal muscle (striated muscle), separated the dorsally located lung from the liver. Therefore, the lung-liver fusion occurred transiently in birds. Rather than the presence of the muscular diaphragm, whether the lung and liver were fused seemed to depend on a timing and sequence of development of the mesothelial coverings of these viscera.

Antialcohol and Hepatoprotective Effects of Tamarind Shell Extract on Ethanol-Induced Damage to HepG2 Cells and Animal Models.

Alcohol liver disease (ALD) is one of the leading outcomes of acute and chronic liver injury. Accumulative evidence has confirmed that oxidative stress is involved in the development of ALD. In this study, we used chick embryos to establish ALD model to study the hepatoprotective effects of tamarind shell exttract (TSE). Chick embryos received 25% ethanol (75 μL) and TSE (250, 500, 750 μg/egg/75 μL) from embryonic development day (EDD) 5.5. Both ethanol and TSE were administrated every two days until EDD15. Ethanol-exposed zebrafish and HepG2 cell model were also employed. The results suggested that TSE effectively reversed the pathological changes, liver dysfunction and ethanol-metabolic enzyme disorder in ethanol-treated chick embryo liver, zebrafish and HepG2 cells. TSE suppressed the excessive reactive oxygen species (ROS) in zebrafish and HepG2 cells, as well as rebuilt the irrupted mitochondrial membrane potential. Meanwhile, the declined antioxidative activity of glutathione peroxidase (GPx) and superoxide dismutase (SOD), together with the content of total glutathione (T-GSH) were recovered by TSE. Moreover, TSE upregulated nuclear factor erythroid 2-related factor 2 (NRF2) and heme oxyense-1 (HO-1) expression in protein and mRNA level. All the phenomena suggested that TSE attenuated ALD through activating NRF2 to repress the oxidative stress induced by ethanol.

Evaluation the effect of Retin - A on some internal organs and skin in pregnant white mice Mus musculus and embryos

The study included the effect of different doses of Ritan A, both in: cream Ritan A topical, and Ritan A drug oral forms. The study included the effect of cream in histopathological lesions of the skin dorsal region and the effect of the drug in causing distortions morphological and histopathological lesions in the liver and kidney of pregnant mice Mus musculus and their fetuses and the extent of its influence in the appearance of congenital malformations macroscopic and influence on the histopathological lesions of the liver and kidney of embryos .&#x0D; Histological examination of the skin of mice treated cream Ritan A showed number of histopathological lesions represented the expansion of the skin layer and the dermis and the occurrence of necrosis of nuclear, and the histological examination of the liver and kidney of pregnant females were treated with a drug Ritan A oral are necrosis and hemorrhage and infiltration of lymphocytes and swelling of the epithelial cells with appearance desquamation of renal tubule epithelium and contraction of renal glomeruli and damage to the glomeruli in the Overdose and lethal dose groups