Fertilized Chicken Eggs Research Articles

Effective technology of disinfecting ozonation of hatching chicken eggs

Relevance. The positive aspects of disinfecting ozonation of hatching eggs allow us to expect successful application of the method not only in large but also in small-scale farms, as well as for experimental and industrial purposes in laboratories and bio-enterprises related to the technological process based on the incubation of a small number of eggs. This determines the importance of expanding the range of ozonizers due to numerous portable devices. There are no clear recommendations for these devices and this causes the need to search for the most effective and harmless modes for the embryo, and schemes of disinfecting ozonation.Methods. The study used fertilized chicken eggs “Hysex Brown” and a portable ozonizer “OZON-OviV”. Ozone concentration 2.0 mg / l. Eggs were treated in a specially made chamber. Technology-1: twice for 30 minutes before incubation and on the 3rd day of incubation. Technology-2: three times for 30 minutes before incubation, on days 3 and 5. The range of studies included: assessment of total microbial contamination (densitometry); identification of microorganisms (MALDI-TOF-spectrometry); biological control of incubation (fertility, hatchability, mortality, developmental abnormalities); assessment of the adequacy of the internal organs (MicroCT); embryo morphometry (weight, length, chest circumference) and calculation of development proportionality indices; histological assessment of the liver.Results. The total ozone concentration during treatment by two methods was 240 mg/l and 360 mg/l, respectively. The disinfecting efficiency of ozonation has been proven, providing a decrease in the level of total microbial contamination by 30% and 40% with double and triple treatment. A tendency to maintain a low total microbial contamination, compared to intact eggs, up to 14 days of incubation has been revealed. The dynamics of the microbial landscape indicate the bacteriostatic effect of ozone in the concentrations used on a wide range of microorganisms. Microtomographic and histological methods confirmed the harmlessness of the technologies used. Along with the more pronounced antibacterial effect of technology-2, the presence of a stimulating effect on the body of the developing embryo was revealed, which determines the preference for its choice.

Critical appraisal of the chorioallantoic membrane model for studying angiogenesis in preclinical research.

Angiogenesis, the biological mechanism by which new blood vessels are generated from existing ones, plays a vital role in growth and development. Effective preclinical screening is necessary for the development of medications that may enhance or inhibit angiogenesis in the setting of different disorders. Traditional in vitro and, in vivo models of angiogenesis are laborious and time-consuming, necessitating advanced infrastructure for embryo culture. A challenge encountered by researchers studying angiogenesis is the lack of appropriate techniques to evaluate the impact of regulators on the angiogenic response. An ideal test should possess reliability, technical simplicity, easy quantifiability, and, most importantly, physiological relevance. The CAM model, leveraging the extraembryonic membrane of the chicken embryo, offers a unique combination of accessibility, low cost, and rapid development, making it an attractive option for angiogenesis assays. This review evaluates the strengths and limitations of the CAM model in the context of its anatomical and physiological properties, and its relevance to human pathophysiological conditions. Its abundant capillary network makes it a common choice for studying angiogenesis. The CAM assay serves as a substitute for animal models and offers a natural setting for developing blood vessels and the many elements involved in the intricate interaction with the host. Despite its advantages, the CAM model's limitations are notable. These include species-specific responses that may not always extrapolate to humans and the ethical considerations of using avian embryos. We discuss methodological adaptations that can mitigate some of these limitations and propose future directions to enhance the translational relevance of this model. This review underscores the CAM model's valuable role in angiogenesis research and aims to guide researchers in optimizing its use for more predictive and robust preclinical studies. The highly vascularized chorioallantoic membrane (CAM) of fertilized chicken eggs is a cost-effective and easily available method for screening angiogenesis, in comparison to other animal models.

An in vivo tumour organoid model based on the chick embryonic chorioallantoic membrane mimics key characteristics of the patient tissue: a proof-of-concept study

BackgroundPatient-derived tumour organoids (PDOs) are highly advanced in vitro models for disease modelling, yet they lack vascularisation. To overcome this shortcoming, organoids can be inoculated onto the chorioallantoic membrane (CAM); the highly vascularised, not innervated extraembryonic membrane of fertilised chicken eggs. Therefore, we aimed to (1) establish a CAM patient-derived xenograft (PDX) model based on PDOs generated from the liver metastasis of a colorectal cancer (CRC) patient and (2) to evaluate the translational pipeline (patient – in vitro PDOs – in vivo CAM-PDX) regarding morphology, histopathology, expression of C-X-C chemokine receptor type 4 (CXCR4), and radiotracer uptake patterns.ResultsThe main liver metastasis of the CRC patient exhibited high 2-[18F]FDG uptake and moderate and focal [68Ga]Ga-Pentixafor accumulation in the peripheral part of the metastasis. Inoculation of PDOs derived from this region onto the CAM resulted in large, highly viable, and extensively vascularised xenografts, as demonstrated immunohistochemically and confirmed by high 2-[18F]FDG uptake. The xenografts showed striking histomorphological similarity to the patient’s liver metastasis. The moderate expression of CXCR4 was maintained in ovo and was concordant with the expression levels of the patient’s sample and in vitro PDOs. Following in vitro re-culturing of CAM-PDXs, growth, and [68Ga]Ga-Pentixafor uptake were unaltered compared to PDOs before transplantation onto the CAM. Although [68Ga]Ga-Pentixafor was taken up into CAM-PDXs, the uptake in the baseline and blocking group were comparable and there was only a trend towards blocking.ConclusionsWe successfully established an in vivo CAM-PDX model based on CRC PDOs. The histomorphological features and target protein expression of the original patient’s tissue were mirrored in the in vitro PDOs, and particularly in the in vivo CAM-PDXs. The [68Ga]Ga-Pentixafor uptake patterns were comparable between in vitro, in ovo and clinical data and 2-[18F]FDG was avidly taken up in the patient’s liver metastasis and CAM-PDXs. We thus propose the CAM-PDX model as an alternative in vivo model with promising translational value for CRC patients.Graphical

BENEFITS OF TEAS AND PROBIOTIC PROFILE AT SPECIFIC STAGES OF EMBRYONIC DEVELOPMEN IN CHICKEN EGGS

Introduction: The Sharr Mountain is characterized by a highly expressed biological diversity, with a particular endemism and a wide spectrum of various types of flora characteristic of this mountainous massif. Plants with nutritional and health benefits can be considered species such as: mountain tea (Sideritis scardica) and St. John's wort (Hypericum perforatum), etc. These bioproducts are characterized by special importance both in terms of their presence, quality, and health benefits. Therefore, exploration of these species would have positive cost for consumption as forms of teas or plant extracts for commercial purposes, but also for health benefits and above all for the population of the country, the region, and beyond. Research Objective: This study aims to investigate the probiotics and the effect of plant extract teas such as mountain tea (Sideritis scardica) and St. John's wort (Hypericum perforatum) by applying a specific dose of extract to chicken embryos at different stages of ontogenesis. Materials and Methods: Fertilized chicken eggs were taken for the study and were incubated in an incubator for 21 days. The eggs were treated with species such as mountain tea (Sideritis scardica) and St. John's wort (Hypericum perforatum) at certain stages of embryonic development. The eggs were opened on the seventh, fourteenth, and twenty-first day. Macroscopic and microscopic changes at certain stages of embryonic development were analyzed using stereomicroscope.The effect of the extract of these teas was studied on the chicken embryos (Gallus domesticus), by applying a specific dose of extract to the chicken embryos at different stages of embryonic development during ontogenesis. Research Results: The dose given in the form of extract has resulted in positive benefits of these teas as effective probiotics during certain stages of embryonic development in chicken embryos. Conclusions: This research will explore the effect of the given dose of plant extracts at certain stages of embryonic development during ontogenesis to observe the benefits of dose administration at specific quantities and times to detect the positive effect of these teas.

MONITORING OF THE LIVING ENVIRONMENT THROUGH AIR POLLUTANTS IN THE POLOG FIELD OF THE REPUBLIC OF NORTH MACEDONIA

Introduction: The Republic of North Macedonia and especially the field pit of Polog in terms of geographical position and climatic aspect is considered as a place with different variables such as: temperature, humidity, radioactive exposure, hilly-mountainous relief. The Sharr mountain in the west and the dry mountain in the north, limit the Polog field pit in all directions, such as north-west and south-east, causing the living environment to deteriorate in the winter period due to air, water and land pollutants. With special emphasis, we highlight the winter season when we have an alarming state of air pollution, such as certain regions of the Republic of North Macedonia and especially the region of the field pit of Polog with the cities of Tetovo and Gostivar and the surrounding area. Recently, the city of Tetova and its surroundings is considered one of the most polluted cities in the country, region and the largest in the world, then this phenomenon of pollution necessarily leads us to unwanted consequences for the population that lives and operates in this region. As a result of environmental factors such as air pollution and other related factors such as: stress, temperature, food, social genes, mental state and other related factors can lead to disorders in the human body and lead to diseases that attack the nervous and cardiovascular systems , skeletal and thyroid. Many studies talk about the alarming condition of patients who are affected by these diseases, therefore our paper will focus on the effect of these factors above all on the actual state of pollutants in the Polog region of North Macedonia. The purpose of the study: This paper aims to investigate the potential causes that can affect the health of the population of the Polog field pit and especially the city of Tetova and its surroundings. As a potential pollutants of the living environment, the following can be considered: carbon monoxide, carbon dioxide, sulfur dioxide, urban combustion from automobiles, combustion of various physical substances, chemical components from factories, smoke from chimneys from burning oil and coal, combustion from wood in the form of pellets, etc. Material and working method: In the study, fertilized chicken eggs were taken and incubated in the incubator for 21 days of incubation. The eggs were treated with different doses of air pollutant extract at certain stages of embryonic development. Using the stereomicroscope with macroscopic and microscopic method were analyzed ontogenetic changes in certain stages of embryonic development. Research results: The results of the research will give a real insight into the situation of the level of pollution of the living environment in the city of Tetovo and its surroundings and in the field pit of Pollog. The monitoring of the living environment can be followed by the treatment of air pollutants in certain doses in the form of ampoule therapy, and the negative effect of the given dose on the hen's eggs will be observed. All eggs with the highest dose of injection had a stagnated embryonic development, whereas egss with lower dose of injections had underdeveloped embryos. Conclusion: From the results of the research we can conclude that the potential pollutants from air pollutants can lead to consequences for the population living in this region but also for the living world in general. These factors can lead to disorders of the nervous system, cardiovascular, and thyroid disorders and other respiratory diseases. This research will give a real insight into the pollution of the living environment and the possible consequences of this at a local, regional and wider level.

THE EFFECT OF THE ANTIBIOTIC GENTAMICIN ON THE STAGES OF ONTOGENESIS IN CHICKEN EGGS

Introduction: Various studies have shown that the effects of the action of drugs, in addition to having positive effects, they can also provoke negative effects in different organs of the body, where by giving more doses in some cases it can exceed the therapeutic limit and become toxic and produce pathological effects in both experimental animals and humans. Uncontrolled use and drug overdose can lead to macroscopic histopathological changes and give teratogenic, mutagenic and carcinogenic effects. Purpose of the paper: This paper aims to study the effects of some drugs at different stages of embryonic development in chicken eggs. The research of this paper will consist of the toxic effect of drugs and macroscopic and microscopic histopathological changes in certain stages of embryonic development of chicken eggs. Material and method: In the study, fertilized chicken eggs were taken and incubated in the incubator for 21 days of incubation. The eggs were treated with the antibiotic gentamicin at certain stages of embryonic development. The eggs hatched on day thirteen, day fifteen, eighteen and day twentyone. Using the stereomicroscope with the macroscopic and microscopic method were analyzed ontogenetic changes in certain stages of embryonic development. Results: The given dose of the antibiotic gentamicin can have negative effects in certain stages of embryonic development in chicken eggs. The dose given in ampullary form in certain quantities to chicken eggs has inhibited and stagnated the processes of embryonic development at different stages compared to control eggs. Conclusion: The given dose of gentamicin can stagnate embryonic development in certain stages and can provoke changes in certain stages of embryonic development. This phenomenon encourages us to do even more detailed research in this direction to see the effect of the given dose and to analyze what consequences it can provoke overdoses at different stages of embryonic development and within the organism. The lethal dose applied at certain stages of ontogenesis may have lethal consequences for the chick embryo at certain stages of embryonic development.

Osteogenic and angiogenic potential of molybdenum-containing mesoporous bioactive glass nanoparticles: An ionic approach to bone tissue engineering

Biomaterials intended for application in bone tissue engineering (BTE) ideally stimulate osteogenesis and angiogenesis simultaneously, as both mechanisms are of critical importance for successful bone regeneration. Mesoporous bioactive glass nanoparticles (MBGNs) can be tailored towards specific biological needs, for example by addition of ions like Molybdenum (Mo). While Mo has been shown to enhance osteogenic differentiation of human bone marrow-derived mesenchymal stromal cells (BMSCs) as well as their ability to form and mature a primitive osseous extracellular matrix (ECM), there are contradictory findings regarding its impact on angiogenesis. In this study, the effects of Mo-MBGNs (mol%: 70 SiO2, 25 CaO, 5 MoO3) on viability, proliferation, osteogenic differentiation, ECM formation and angiogenic response of BMSCs were compared to undoped MBGNs (in mol%: 70 SiO2, 30 CaO) and a control group of BMSCs. Furthermore, a human umbilical vein endothelial cells tube formation assay and a chorioallantoic membrane-assay using fertilized chicken eggs were used to analyze angiogenic properties. Mo-MBGNs were cytocompatible and promoted the proliferation of BMSCs. Furthermore, Mo-MBGNs showed promising osteogenic properties as they enhanced osteogenic differentiation, ECM formation and maturation as well as the gene expression and protein production of relevant osteogenic factors in BMSCs. However, despite the promising outcome on osteogenic properties, the addition of Mo to MBGNs resulted in anti-angiogenic effects. Due to the high relevance of vascularization in-vivo, the anti-angiogenic properties of Mo-MBGNs might hamper their osteogenic properties and therefore might restrict their performance in BTE applications. These limitations can be overcome by the addition of ions with distinct pro-angiogenic properties to the Mo-MBGNs-composition. Due to their promising osteogenic properties, Mo-MBGNs constitute a suitable basis for further research in the field of ionic (growth factor free) BTE.

Chick Embryo Chorioallantoic Membrane as a Platform for Assessing the InVivo Efficacy of Chimeric Antigen Receptor T-cell Therapy in Solid Tumors.

The fertilized chicken egg chorioallantoic membrane (CAM), a highly vascularized membrane nourishing the developing embryo, also supports rapid growth of three-dimensional vascularized tumors from engrafted cells and tumor explants. Because murine xenograft models suffer limitations of time, cost, and scalability, we propose CAM tumors as a rapid, efficient screening tool for assessing anti-tumor efficacy of chimeric Ag receptor (CAR) T cells against solid tumors. We tested the efficacy of human epidermal growth factor receptor 2 (HER2)-specific CAR T cells against luminescent, HER2-expressing (FaDu, SCC-47) or HER2-negative (MDA-MB-468) CAM-engrafted tumors. Three days after tumor engraftment, HER2-specific CAR T cells were applied to tumors grown on the CAM. Four days post-CAR T cell treatment, HER2-expressing FaDu and SCC-47 tumors treated with CAR T showed reduced viable cancer cells as assessed by luciferase activity. This reduction in viable tumor cells was confirmed by histology, with lower Ki-67 staining observed in CAR T cell-treated tumors relative to T cell-treated controls. Persistence of CAR T in CAM and tumor tissue 4 days post-treatment was confirmed by CD3 staining. Altogether, our findings support further development of the chick CAM as an invivo system for rapid, scalable screening of CAR T cell efficacy against human solid tumors.

Developmental 6:2 FTCA exposure impairs renal development in chicken embryos via IGF signaling

6:2 fluorotelomer carboxylic acid (6:2 FTCA) is a perfluorooctanoic acid (PFOA) substitute, which is supposedly less accumulative and toxic than PFOA. However, 6:2 FTCA is structurally similar to PFOA, and there had already been reports about its toxicities comparable to PFOA. The aim of the current study is to assess potential effects of developmental exposure to 6:2 FTCA on the development of kidney in chicken embryo and to investigate underlying mechanism. Fertile chicken eggs were exposed to 1.25 mg/kg, 2.5 mg/kg or 5 mg/kg doses of 6:2 FTCA, or 2 mg/kg PFOA, then incubated to hatch. Serum and kidney of hatchling chickens were collected. Blood urea nitrogen (BUN) and creatinine (Cre) levels were measured with commercially available kits. Morphology of kidney was assessed with histopathology. To further reveal molecular mechanism of observed endpoints, IGF signaling molecules were assessed in the kidney samples with qRT-PCR, results indicated that IGFBP3 is a potentially crucial molecule. Lentiviruses overexpressing or silencing IGFBP3 were designed and applied to enhance/suppress the expression of IGFBP3 in developing chicken embryo for further verification of its role in the observed effects. Disrupted nephron formation, in the manifestation of decreased glomeruli number/area and increased serum BUN/Cre levels, was observed in the animals developmentally exposed to 6:2 FTCA. Correspondingly, IGF signaling molecules (IGF1, IGF1R and IGFBP3) were affected by 6:2 FTCA exposure. Meanwhile, overexpression of IGFBP3 effectively alleviated such changes, while silencing of IGFBP3 mimicked observed effects. In conclusion, developmental exposure to 6:2 FTCA is associated with disrupted chicken embryo renal development, in which IGFBP3 seems to be a remarkable contributor, suggesting potential health risks for human and other species. Further risk assessments and mechanistic works are necessary.

Development of Recombinant PLC-Zeta Protein as a Therapeutic Intervention for the Clinical Treatment of Oocyte Activation Failure.

The sperm-specific phospholipase C zeta (PLCζ) protein is widely considered as the predominant physiological stimulus for initiating the Ca2+ release responsible for oocyte activation during mammalian fertilization. The increasing number of genetic and clinical reports that directly link PLCζ defects and/or deficiencies with oocyte activation failure (OAF) necessitates the use of a powerful therapeutic intervention to overcome such cases of male factor infertility. Currently, in vitro fertilization (IVF) clinics treat OAF cases after intracytoplasmic sperm injection (ICSI) with Ca2+ ionophores. Despite their successful use, such chemical agents are unable to trigger the physiological pattern of Ca2+ oscillations. Moreover, the safety of these ionophores is not yet fully established. We have previously demonstrated that recombinant PLCζ protein can be successfully used to rescue failed oocyte activation, resulting in efficient blastocyst formation. Herein, we produced a maltose binding protein (MBP)-tagged recombinant human PLCζ protein capable of inducing Ca2+ oscillations in mouse oocytes similar to those observed at fertilization. Circular dichroism (CD) experiments revealed a stable, well-folded protein with a high helical content. Moreover, the recombinant protein could retain its enzymatic properties for at least up to 90 days after storage at -80 °C. Finally, a chick embryo model was employed and revealed that exposure of fertilized chicken eggs to MBP-PLCζ did not alter the embryonic viability when compared to the control, giving a first indication of its safety. Our data support the potential use of the MBP-PLCζ recombinant protein as an effective therapeutic tool but further studies are required prior to its use in a clinical setting.

Effect of in ovo feeding of xylobiose and xylotriose on plasma immunoglobulin, cecal metabolites production, microbial ecology, and metabolic pathways in broiler chickens

BackgroundDietary supplementation of xylooligosaccharides (XOS) has been found to influence gut health by manipulating cecal microbiota and producing microbe-origin metabolites. But no study investigated and compared the effect of in ovo feeding of xylobiose (XOS2) and xylotriose (XOS3) in chickens. This study investigated the effect of in ovo feeding of these XOS compounds on post-hatch gut health parameters in chickens. A total of 144 fertilized chicken eggs were divided into three groups: a) non-injected control (CON), b) XOS2, and c) XOS3. On the 17th embryonic day, the eggs of the XOS2 and XOS3 groups were injected with 3 mg of XOS2 and XOS3 diluted in 0.5 mL of 0.85% normal saline through the amniotic sac. After hatching, the chicks were raised for 21 d. Blood was collected on d 14 to measure plasma immunoglobulin. Cecal digesta were collected for measuring short-chain fatty acids (SCFA) on d 14 and 21, and for microbial ecology and microbial metabolic pathway analyses on d 7 and 21.ResultsThe results were considered significantly different at P < 0.05. ELISA quantified plasma IgA and IgG on d 14 chickens, revealing no differences among the treatments. Gas chromatography results showed no significant differences in the concentrations of cecal SCFAs on d 14 but significant differences on d 21. However, the SCFA concentrations were lower in the XOS3 than in the CON group on d 21. The cecal metagenomics data showed that the abundance of the family Clostridiaceae significantly decreased on d 7, and the abundance of the family Oscillospiraceae increased on d 21 in the XOS2 compared to the CON. There was a reduction in the relative abundance of genus Clostridium sensu stricto 1 in the XOS2 compared to the CON on d 7 and the genus Ruminococcus torques in both XOS2 and XOS3 groups compared to the CON on d 21. The XOS2 and XOS3 groups reduced the genes for chondroitin sulfate degradation I and L-histidine degradation I pathways, which contribute to improved gut health, respectively, in the microbiome on d 7. In contrast, on d 21, the XOS2 and XOS3 groups enriched the thiamin salvage II, L-isoleucine biosynthesis IV, and O-antigen building blocks biosynthesis (E. coli) pathways, which are indicative of improved gut health. Unlike the XOS3 and CON, the microbiome enriched the pathways associated with energy enhancement, including flavin biosynthesis I, sucrose degradation III, and Calvin-Benson-Bassham cycle pathways, in the XOS2 group on d 21.ConclusionIn ovo XOS2 and XOS3 feeding promoted beneficial bacterial growth and reduced harmful bacteria at the family and genus levels. The metagenomic-based microbial metabolic pathway profiling predicted a favorable change in the availability of cecal metabolites in the XOS2 and XOS3 groups. The modulation of microbiota and metabolic pathways suggests that in ovo XOS2 and XOS3 feeding improved gut health during the post-hatch period of broilers.

Sevofluran Maruziyetinin Civciv Embriyo Gelişimi Üzerindeki Ex-Ovo Değerlendirilmesi: Anjiyogenez Etkilerinin Araştırılması

Objective: Angiogenesis, defined as the formation of new blood vessels from pre-existing ones, plays a vital role in both physiological and pathological conditions. Understanding agents that can influence angiogenesis is crucial in managing diseases where angiogenesis is dysregulated. This study focuses on sevoflurane, a commonly used inhalation anesthetic, whose effects on angiogenesis and embryonic development are not well understood. Our objective was to assess the impact of sevoflurane exposure on angiogenesis using the ex-ovo chick chorioallantoic membrane model. Methods: In this model, fertilized chicken eggs were exposed to sevoflurane at concentrations of 2% and 4%, for varying durations of 1, 2, and 4 hours. The embryos were then divided into control and experimental groups for quantitative angiogenesis assessment using Image J software, followed by statistical analysis with one-way analysis of variance. Results: The results indicate that sevoflurane exposure has a dose-dependent positive effect on angiogenesis, with significant increases in vascular density observed in embryos exposed to both concentrations compared to the control group. Additionally, the length of exposure was found to further enhance these angiogenic effects. Conclusion: Despite the dose and duration-dependent impact of sevoflurane on angiogenesis, the existing literature presents mixed findings, highlighting the need for additional research to elucidate sevoflurane’s role in angiogenesis. This is particularly important for understanding its implications in various medical conditions, such as cancer, wound healing, and fetal development. Future investigations into sevoflurane’s effects on placental angiogenesis could also provide valuable insights into its potential consequences on intrauterine growth. Keywords: Angiogenesis, sevoflurane, chicken embryo, model organisms, ex-ovo

Assessment of Antiangiogenic and Cytotoxic Effects of Moringa oleifera Silver Nanoparticles Using Cell Lines

Angiogenesis is the physiological process through which new blood vessels are developed from pre-existing vessels. It includes steps like migration, differentiation, and growth of endothelial cells. Furthermore, it plays a critical role in cancer formation and metastasis. Recently, Moringa oleifera (MO) has gained interest due to its properties in various fields, particularly in nanoparticle (NP) technology. This study aims to demonstrate the cytotoxic and antiangiogenic effects of MO silver nanoparticles (MO-AgNPs). Briefly, the cytotoxic effects of MO-AgNPs on Ea. hy 926 and HT 29 cell lines were observed in the Colony Formation Assay. The antiproliferative effects of MO-AgNPs were then further confirmed using MTT assay, in addition to morphological characterization, migration and closure percent calculation. To evaluate the antiangiogenic effect of MO, the chorioallantoic membrane (CAM) assay was done. Fertilized chicken eggs were divided into three groups: MO-AgNPs, MO ethanol extract and negative control. The results showed antiangiogenic effects were observed at 6-12 µg/mL concentration of MO-AgNPs. Whereas the plate efficiency was 45.7 % ± 1%. The surviving fraction at 12, 6, 3, and 1.5 μg/mL was 27.3± 0.5%, 30.1 ± 0.5 %, 41.8 ± 1.5%, and 69.8 ± 2.5 %, respectively. MO-AgNPs showed higher cytotoxic activity than MO ethanol extract on the Ea. hy926 cell line, where the IC50 of MO-AgNPs were 74, 35, and 12 µg/mL for 24, 48 and 72 hours, respectively. On the other hand, the IC50 of MO ethanol extract was above 200, 135 and 11 µg/mL for 24, 48 and 72 hours, respectively. Additionally, significant morphological changes were observed in Ea. hy cells. Whereas the closure percent average of Ea. hy cells at 1.5, 3, 6, and 12 µg/mL were 91.603, 88.507, 84.599, and 63.144%, respectively. Based on the results showed in this study, MO extract shows potential to be an effective anti-angiogenic agent and is a promising therapeutic option in cancer treatment.

Histomorphometrical and histochemical study of small intestine development in local chicken (Gallus gallus domesticus) and duck (Anas platyrhynchos domesticus) embryos

This study demonstrated for the first time the histomorphometry of the small intestine (SI) and distribution of goblet and Paneth cells, collagen, and smooth muscle fibers in (SI) of the local chicken and duck during embryological development using the combined PAS and Alcian blue (pH2.5), Geimsa, and Masson's trichrome stains. One hundred and four fertilized chicken eggs and one hundred and forty fertilized duck eggs were collected from Mosul city. The sample placed in an incubator with automatic movement, ventilation, and humidity was (60%) and the temperature was (37.7 °C) for chicken and (37.5 °C) for duck. The histomorphometry showed significant differences between chicken and duck embryos at the 19th and 21st days of incubation of the villi's length, width, and apparent surface area. As for the epithelium height and the intestinal crypts, there were significant differences between chicken and duck embryos at the 18th and 20th days of incubation in the ileum and jejunum. Furthermore, the mucosa was thicker in chicken embryos in the day of hatching, the submucosa was thicker in ducks during the 18th day of the incubation in the duodenum. During day 15 of the incubation in chicken embryos in the jejunum and ileum, the muscularis layer was thicker in the duodenum and jejunum and less thick in the ileum, and the collagen fibers were less too in the duck embryos at hatching d. The serosa was thicker in the duck embryos. The percentage of the goblet cells was higher with a larger density in duck embryos than chicken at hatching day. Paneth cells were characterized by acidic granular cytoplasm. The achieved outcomes of histomorphometry and the density variances of the glycoprotein secretion are probably linked with numerous definite roles of (SI) parts in the progressions of nutrient absorption.

Abstract 6900: Patient-derived xenografts from circulating cancer stem cells and their potential utility in personalized medicine of pancreatic cancer

Abstract Background: Pancreatic cancer carries a poor prognosis in the majority of patients, with an overall 5-year survival rate of less than 10%. Therefore, the development of new therapies and preclinical models is crucial and urgently needed. Circulating cancer stem cells (cCSCs) are a subset of circulating cancer cells that have stem cell-like properties and play a significant role in tumor progression, metastasis, and treatment resistance. Patient-derived xenografts (PDX) reflect the genotypic and phenotypic landscape of patients' cancers and are useful for drug screening, biomarker development, and preclinical evaluation of personalized medicine strategies. In this study, we describe a fast, simple, and cost effective method to generate PDX on Chorioallantoic Membrane (CAM) from circulating cancer stem cells. Methods: In this study, primary cultures from circulating cancer stem cells were established using sphere-forming assays from 30 patients with newly diagnosed pancreatic cancer. Subsequently, tumorspheres were transplanted onto the CAM membrane of fertilized chicken eggs to form patient- derived xenografts. Results: For the identification of cCSCs from peripheral blood of pancreatic cancer patients, an innovative in vitro sphere-forming assay was established. The number of tumorspheres was correlated with clinicopathological parameters like tumor size, lymph node involvement and grading status of the primary tumor. Implantation of tumorspheres onto the CAM was successful in ~90 % of the applied samples. The histological analysis suggests that the PDX tumors were morphologically and pathologically identical to primary tumor tissue. Conclusions: The number of tumorspheres was associated with clinicopathological parameters. Further follow-up is needed to evaluate the prognostic significance of tumorsphere detection in pancreatic cancer. Generation of PDX from cCSCs on CAM has a high success rate and may become an integral part of the drug development arena, including drug screening and biomarker development. Citation Format: Monika Pizon, Dorothea Schott, Andreas Ettner-Sittner, Benedikt J. Wagner, Katharina Pachmann, Thiha Aung, Christina Hackl, Silke Haerteis. Patient-derived xenografts from circulating cancer stem cells and their potential utility in personalized medicine of pancreatic cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6900.

Determination of Lethal Dose of Disodium 5’ Ribonucleotide (E635) on Embryonic Development of &lt;i&gt;Gallus gallus&lt;/i&gt;

Disodium 5’ Ribonucleotide (E635), a food additive, has FDA safety, but concerns about its potential toxicity in developmental biology have been raised due to limited research on its lethal effects. The present research aimed to investigate the lethal dose of E635 when induced into a vertebrate chick embryo model. Pilot doses of E635 ranging from 1 μg- 400 μg per egg were inoculated. Consequently, fertilized chick eggs were randomly assigned to 6 groups, including 1 control and 5 differently intoxicated groups of E635 doses per egg. E635 was induced in eggs by in vivo administration and incubated for up to 15 days. LD50 was determined by using probit analysis in SPSS. The study showed morphological, physiological and survival alterations. A dose-dependent mortality was observed by E635 induction in the present study. The results were interpreted for functional and biological changes in the developing embryo of Gallus gallus to confirm the induced toxicity. LD50 of E635 on chick embryo was found to be 0.054 μg per egg.

An Adaptive Partial Least-Squares Regression Approach for Classifying Chicken Egg Fertility by Hyperspectral Imaging.

Partial least-squares (PLS) regression is a well known chemometric method used for predictive modelling, especially in the presence of many variables. Although PLS was not initially developed as a technique for classification tasks, scientists have reportedly used this approach successfully for discrimination purposes. Whereas some non-supervised learning approaches, including, but not limited to, PCA and k-means clustering, do well in identifying/understanding grouping and clustering patterns in multidimensional data, they are limited when the end target is discrimination, making PLS a preferable alternative. Hyperspectral imaging data on a total of 672 fertilized chicken eggs, consisting of 336 white eggs and 336 brown eggs, were used in this study. Hyperspectral images in the NIR region of the 900-1700 nm wavelength range were captured prior to incubation on day 0 and on days 1-4 after incubation. Eggs were candled on incubation day 5 and broken out on day 10 to confirm fertility. While a total number of 312 and 314 eggs were found to be fertile in the brown and white egg batches, respectively, the total number of non-fertile eggs in the same set of batches was 23 and 21, respectively. Spectral information was extracted from a segmented region of interest (ROI) of each hyperspectral image and spectral transmission characteristics were obtained by averaging the spectral information. A moving-thresholding technique was implemented for discrimination based on PLS regression results on the calibration set. With true positive rates (TPRs) of up to 100% obtained at selected threshold values of between 0.50 and 0.85 and on different days of incubation, the results indicate that the proposed PLS technique can accurately discriminate between fertile and non-fertile eggs. The adaptive PLS approach was, thereby, presented as suitable for handling hyperspectral imaging-based chicken egg fertility data.

Optical luminescent properties of eggs

Relevance. Optical photoluminescent properties of poultry eggs can serve as indicators of their quality both for incubation processes and for food technology processes. At the initial stage, it is important to evaluate the spectral characteristics of excitation and luminescence, as well as the parameters of table and incubation eggs of different quality and with different shell color.Methods. Chicken eggs of Chinese silk and Hysex brown crosses, as well as samples of duck, goose and quail eggs were used for research. The measurements were carried out on a diffraction spectrofluorimeter “Fluorat02-Panorama” with built-in software “Panorama Pro”. The obtained spectral characteristics were used to calculate the energy (integral absorption capacity and luminescence flux, spectrum energy) and statistical parameters (expectation, variance, asymmetry, kurtosis), as well as the Stokes shift.Results. Photoluminescent properties of poultry eggs are manifested in the ultraviolet region of the spectrum. Qualitatively, the spectra of different species of domestic birds are similar with a slight difference in the Stokes shift. For fertilized chicken eggs, the photoluminescence flux is 2.7 times greater than that of unfertilized eggs, while the spread of values for unfertilized eggs is 5.6 times less. Spectrum parameters such as asymmetry and kurtosis differ significantly for the luminescence of dark and light-shell chicken eggs. For table eggs, the integral energy parameters are 2.8–3.1 times greater than those of incubation eggs.

Determination of the embryotoxic effects of propofol injected into eggs on the cerebellum and spinal cord using histologic methods: an animal study.

This study aims to determine the possible embryotoxic effects of propofol on the cerebellum and spinal cord using fertile chicken eggs. A total of 430 fertile eggs were divided into 5 groups: control, saline, 2.5 mg.kg-1, 12.5 mg.kg-1, and 37.5 mg.kg-1 propofol. Injections were made immediately before incubation via the air chamber. On the 15th, 18th, and 21st day of incubation, 6 embryos from each group were evaluated. Serial paraffin sections taken from the cerebellum and spinal cord were stained with hematoxylin-eosin, Kluver-Barrera, toluidine blue, and periodic acid-Schiff's reaction. The outer granular layer and total cortex thickness were measured, and the linear density of the Purkinje cells was determined. The ratios of the substantia grisea surface area to the total surface area of the spinal cord were calculated. The transverse and longitudinal diameters of the canalis centralis were also assessed. No structural malformation was observed in any embryos examined macroscopically. No significant difference was observed between the groups in terms of development and histologic organization of the cerebellum and spinal cord. However, on the 15th, 18th, and 21st day, the outer granular layer (p < 0.001 for all days) and the total cortex thickness (p < 0.01, p < 0.001, and p < 0.001, respectively) decreased significantly in different propofol dose groups in varying degrees in the cerebellum. Similarly, in the spinal cord, there were significant changes in the ratios of the substantia grisea surface area to the total surface area (p < 0.01 and p < 0.001, respectively). It was concluded that the in-ovo-administered propofol given immediately before incubation has adverse effects on the developing cerebellum and spinal cord. Therefore, it is important for anesthesiologists always to remain vigilant when treating female patients of childbearing age.

Determinación del efecto embriotóxico del Metronidazol mediante un modelo in ovo

Metronidazole is an imidazole group bactericidal antibiotic used against anaerobic bacteria and some protozoa. There is no detailed information about the embryotoxic effect of Metronidazole. This study aims to determine the embryotoxic activity of Metronidazole using an in ovo method. A total of 210 fertile chicken eggs were placed in an incubator, divided into seven equal groups of 30. The first group was considered as the control group. On the seventh day of the study, Metronidazole was administered to the other six groups within 50 µL saline solution at doses of 250, 125, 62.5, 31.25, and 15.62 µg·egg-1 (5; 2.5; 1.25; 0.625; 0.312 mg·kg-1). At the end of the incubation period, the eggs hatched, and the number of live and dead embryos was determined. There were no significant differences in deaths between the groups (P&gt;0.05 in all cases). No anomaly was detected in the macroscopic morphology of the embryos. As a result, it can be stated that Metronidazole may be safe for use during pregnancy. However, it is necessary to conduct molecular and histopathological studies to investigate the effects of this drug on organogenesis, especially in mammalian embryos.