THE SURVIVAL RATE OF CHICK EMBRYO INFLUENCED BY NICOTINE AND ANTIOXIDANT GREEN TEA (CAMELLIA SINENSIS)

Objectives: To study the effects of Nicotine on the survival rate of chick embryo and to evaluate the preventive role by the antioxidant green tea camellia sinensis. Study Design: Laboratory-based experimental study. Place and Duration of Study: Army Medical College, Rawalpindi, from Nov 2011 to Nov 2012. Methodology: A total of 75 Fayoumi fresh fertilized eggs were kept at Poultry Research Institute, Rawalpindi. Experimental solutions were injected to the eggs of four groups at forty-eight hours of incubation and the effect on the growth was recorded and compared with the control group. The control and experimental groups were observed to assess the effects of Nicotine and the role of antioxidant green tea Camellia sinensis on the survival rate and growth of chick embryo’s femur. Results: The control group (group-1) showed 100% survival rate of embryos. The experimental group-2 showed survival rate of 100% embryos and the group-3 and group-4 showed survival rate 50% and 70% of embryos, respectively. Conclusion: It was concluded that green tea decreases the oxidative stress caused by the Nicotine.

Morphological, Histological Changes and Acetyl Cholinesterase Activity in Chick Embryos After Exposure to Abamectin Insecticide

Abamectin is a bio-insecticide, derived from the soil bacteria Streptomyces avermitilis. This insecticide is used in public health and agriculture to protect crops. Major adverse impacts of Abamectin are neurological symptoms acting on the peripheral nervous system. The aim of the current study is to reveal the toxic effects of Abamectin on chick embryo Ross 308 including morphological and histological changes and acetyl cholinesterase activity. 120 fresh fertilized eggs were divided into 6 groups; two of them were used as control. After 2 days of incubation, the eggs were injected with 100 µL of Abamectin solution (diluted at concentrations 360, 540, 900 and 1800 ppm) into the yolk sac. The Results showed that the mortality increased significantly in chicks treated with Abamectin, but had a lower weight in comparison to the control groups. Treated chicks started hatching at day 22 but were physically weak with drooping limbs, paralysis and then died after 24 hours of hatching. Some chicks did not normally hatch and needed assistance. They characterized by limb defects, failure retraction of yolk sac with bleeding. Histological examination of the liver showed hepatic cell degeneration, congestion in the central vein, infiltration of inflammatory cells and hepatocytes necrosis. Furthermore, the Acetyl cholinesterase enzyme analysis showed a significant decrease in the enzyme activity which leads to inhibition the activity of the body systems. It is concluded that low and high concentration of Abamectin has adverse impacts on chick embryo by changing some of morphological, histological characteristics and acetyl cholinesterase activity.

Chick Embryo Experimental Platform for Micrometastases Research in a 3D Tissue Engineering Model: Cancer Biology, Drug Development, and Nanotechnology Applications

Colonization of distant organs by tumor cells is a critical step of cancer progression. The initial avascular stage of this process (micrometastasis) remains almost inaccessible to study due to the lack of relevant experimental approaches. Herein, we introduce an in vitro/in vivo model of organ-specific micrometastases of triple-negative breast cancer (TNBC) that is fully implemented in a cost-efficient chick embryo (CE) experimental platform. The model was built as three-dimensional (3D) tissue engineering constructs (TECs) combining human MDA-MB-231 cells and decellularized CE organ-specific scaffolds. TNBC cells colonized CE organ-specific scaffolds in 2–3 weeks, forming tissue-like structures. The feasibility of this methodology for basic cancer research, drug development, and nanomedicine was demonstrated on a model of hepatic micrometastasis of TNBC. We revealed that MDA-MB-231 differentially colonize parenchymal and stromal compartments of the liver-specific extracellular matrix (LS-ECM) and become more resistant to the treatment with molecular doxorubicin (Dox) and Dox-loaded mesoporous silica nanoparticles than in monolayer cultures. When grafted on CE chorioallantoic membrane, LS-ECM-based TECs induced angiogenic switch. These findings may have important implications for the diagnosis and treatment of TNBC. The methodology established here is scalable and adaptable for pharmacological testing and cancer biology research of various metastatic and primary tumors.

A novel role for the extraembryonic area opaca in positioning the primitive streak of the early chick embryo

Classical studies have established that the marginal zone, a ring of extraembryonic epiblast immediately surrounding the embryonic epiblast (area pellucida) of the chick embryo is important in setting embryonic polarity by positioning the primitive streak, the site of gastrulation. The more external extraembryonic region (area opaca) was only thought to have nutritive and support functions. Using experimental embryology approaches, this study reveals three separable functions for this outer region: first, juxtaposition of the area opaca directly onto the area pellucida induces a new marginal zone from the latter; this induced domain is entirely posterior in character. Second, ablation and grafting experiments using an isolated anterior half of the blastoderm and pieces of area opaca suggest that the area opaca can influence the polarity of the adjacent marginal zone. Finally, we show that the loss of the ability of such isolated anterior half-embryos to regulate (re-establish polarity spontaneously) at the early primitive streak stage can be rescued by replacing the area opaca by one from a younger stage. These results uncover new roles of chick extraembryonic tissues in early development.

Supplemental L-Arginine Improves the Embryonic Intestine Development and Microbial Succession in a Chick Embryo Model

Early colonization of intestinal microbiota plays an important role in intestinal development. However, the microbial succession at an embryonic stage and its assembly patterns induced by prenatal nutrition are unknown. In the present study, we used a chick embryo model to investigate the effects of in ovo feeding (IOF) of L-arginine (Arg) on the intestinal development and microbial succession of embryos. A total of 216 fertile eggs were randomly distributed into 2 groups including the non-injected control group and IOF of Arg group with 7 mg/egg. The results showed that IOF Arg increased the intestinal index, absolute weight of jejunum, and improved jejunal morphology in terms of villus width and surface area (p < 0.05). The relative mRNA expressions of mTOR and 4E-BP1 were up-regulated and accompanied by higher contents of Mucin-2 in the Arg group (p < 0.05). There was a significant elevation in contents of serum glucose and high-density lipoprotein cholesterol, whereas there was a decreased low-density lipoprotein cholesterol in the Arg group (p < 0.05). Additionally, Proteobacteria and Firmicutes were major intestinal bacteria species at the embryonic stage. However, Arg supplementation targeted to shape assembly patterns of microbial succession and then changed microbial composition (p = 0.05). Meanwhile, several short-chain fatty acids (SCFAs)-producing bacteria, such as Roseburia, Blautia, and Ruminococcus were identified as biomarkers in the Arg group (LDA > 3, p < 0.05). Accordingly, significant elevated concentrations of SCFAs, including lactic acid and formic acid, were observed in the Arg group (p < 0.05), accompanied by the higher concentration of butyric acid (0.05 < p < 0.10). In conclusion, prenatal Arg supplementation improved embryonic intestine development by regulating glucose and lipid homeostasis to supply more energy for chick embryos. The possible mechanism could be the roles of Arg in shaping the microbial assembly pattern and succession of the embryonic intestine, particularly the enrichment of potential probiotics. These findings may contribute to exploring nutritional strategies to establish health-promoting microbiota by manipulating prenatal host-microbe interactions for the healthy development of neonates.

Reconstruction of distinct vertebrate gastrulation modes via modulation of key cell behaviours in the chick embryo

The morphology of gastrulation driving the internalisation of the mesoderm and endoderm differs dramatically among vertebrate species. It ranges from involution of epithelial sheets of cells through a circular blastopore in amphibians to ingression of mesenchymal cells through a primitive streak in amniotes. By targeting signalling pathways controlling critical cell behaviours in the chick embryo, we generated crescent- and ring-shaped mesendoderm territories in which cells can or cannot ingress. These alterations subvert the formation of the chick primitive streak into the gastrulation modes seen in amphibians, reptiles and teleost fish. Our experimental manipulations are supported by a theoretical framework linking cellular behaviors to self-organized multi-cellular flows in the accompanying paper. All together, this suggests that the evolution of gastrulation movements are largely determined by the shape of and cell behaviours in the mesendoderm territory across different species, and controlled by a relatively small number of signalling pathways.