Fertilized Chicken Eggs Research Articles

Knock-down of oncohistone H3F3A-G34W counteracts the neoplastic phenotype of giant cell tumor of bone derived stromal cells

Giant cell tumors of bone (GCTB) are semi-malignant tumors associated with extensive osteolytic defects and massive bone destructions. They display a locally aggressive behavior and a very high recurrence rate. Recently, a single mutation has been identified in GCTB affecting the H3F3A gene coding for the histone variant H3.3 (H3.3-G34W). The aim of this study was to investigate whether H3.3-G34W is sufficient to drive tumorigenesis in GCTB. Initially, we confirmed the high frequency of this mutation in 94% of 84 analyzed tissue samples. Using a siRNA based approach we could selectively knockdown H3.3-G34W in primary neoplastic stromal cells isolated from tumor tissue (GCTSC). H3.3-G34W knockdown caused a significant inhibition of cell proliferation, migration and colony formation capacity in vitro. Xenotransplantation of GCTSCs onto the chorioallantoic membrane of fertilized chicken eggs further demonstrated a significant impact of H3.3-G34W knockdown on tumor engraftment and growth in vivo. Our data indicate that H3.3-G34W is sufficient to drive tumorigenesis in GCTB. Apart from the application of H3.3-G34W screening as diagnostic tool, our data suggest that H3.3-G4W represents a promising target for the development of new GCTB therapies.

Antiangiogenic potential of Jatropha curcas latex in the chick chorioallantoic membrane model

AIMS: To perform a physicochemical and phytochemical characterization of Jatropha curcas latex and to investigate its antiangiogenic potential. METHODS: We performed an initial physicochemical characterization of J. curcas latex using thermal gravimetric analyses and Fourier Transform Infrared spectroscopy. After that, phenols, tannins and flavonoids were quantified. Finally, the potential of J. curcas latex to inhibit angiogenesis was evaluated using the chick chorioallantoic membrane model. Five groups of 20 fertilized chicken eggs each had the chorioallantoic membrane exposed to the following solutions: (1) water, negative control; (2) dexamethasone, angiogenesis inhibitor; (3) Regederm®, positive control; (4) 25% J. curcas latex diluted in water; (5) 50% J. curcas latex diluted in water; and (6) J. curcas crude latex. Analysis of the newly-formed vascular net was made through captured images and quantification of the number of pixels. Histological analyses were performed to evaluate the inflammation, neovascularization, and hyperemia parameters. The results were statically analyzed with a significance level set at p ˂0.05.RESULTS: Physicochemical characterization showed that J. curcas latex presented a low amount of cis-1.4-polyisoprene, which reduced its elasticity and thermal stability. Phytochemical analyses of J. curcas latex identified a substantial amount of phenols, tannins, and flavonoids (51.9%, 11.8%, and 0.07% respectively). Using a chick chorioallantoic membrane assay, we demonstrated the antiangiogenic potential of J. curcas latex. The latex induced a decrease in the vascularization of the membranes when compared with neutral and positive controls (water and Regederm®). However, when compared with the negative control (dexamethasone), higher J. curcas latex concentrations showed no significant differences.CONCLUSIONS: J. curcas latex showed low thermal stability, and consisted of phenols, tannins, and flavonoids, but little or no rubber. Moreover, this latex demonstrated a significant antiangiogenic activity on a chick chorioallantoic membrane model. The combination of antimutagenic, cytotoxic, antioxidant and antiangiogenic properties makes J. curcas latex a potential target for the development of new drugs.

Hen's Egg Test for Micronucleus Induction (HET-MN).

The classical in vitro genotoxicity test battery is known to be sensitive for indicating genotoxicity. However, a high rate of "misleading" positives was reported when three assays were combined as required by several legislations. Despite the recent optimizations of the standard in vitro tests, two gaps could merely be addressed with assays based on monolayer cell cultures, that is, the route of exposure and a relevant intrinsic metabolic capacity to transform chemicals into reactive metabolites. Following these considerations, fertilized chicken eggs have been introduced into genotoxicity testing and were combined with a classical readout parameter, i.e., the analysis of micronucleus frequency in erythrocytes, to develop the hen's egg test for micronucleus induction, the HET-MN. As a major advantage the test mirrors the systemic availability of compounds after oral exposure reflecting certain steps of ADME without being considered as an animal experiment. After a successful validation exercise the detailed protocol is given here.

Cardiac Enlargement in the Chick Embryo Induced by Hypothermic Incubation Is Due to a Combination of Hyperplasia and Hypertrophy of Cardiomyocytes

Hypothermic incubation of chicken eggs leads to smaller embryos with enlarged hearts, originally described as hypertrophic. Over the years, however, accumulated evidence suggested that hyperplasia, rather than hypertrophy, is the predominant mechanism of cardiac growth during the prenatal period. We have thus set to re-evaluate the hypothermia model to precise the exact cellular mechanism behind cardiac enlargement. Fertilized chicken eggs were incubated at either 37.5 °C (normothermia) or 33.5 °C from embryonic day (ED) 13 onward (hypothermia). Sampling was performed at ED17, at which point wet embryo and heart weight were recorded, and the hearts were submitted to histological examination. In agreement with previous results, the hypothermic embryos were 29% smaller and had hearts 18% larger, translating into a 67% increase in the heart to body weight ratio (P < 0.05 for all parameters). The cell size was essentially the same between control and hypothermic hearts in all regions analysed. Likewise, there was no significant relationship between the cell size and heart weight; however, in the hypothermic hearts, there was a trend showing positive correlation between cell sizes in different cardiac regions and heart weight. Proliferation rate, determined on the basis of anti-phosphohistone H3 immunofluorescence, showed an overall increase in the hypothermic group, reaching statistical significance (P = 0.02, t-test) in the right ventricle. The proliferation rate was similar among different regions of the same heart. However, the correlation between the proliferation rate and heart weight was only small (r2 = 0.007 and r2 = 0.234 for the normothermic and hypothermic group, respectively). We thus conclude that hyperplasia is the predominant response mechanism in this volume-overload model; mechanistically, decreased heart rate at lower temperature increases the end-diastolic and stroke volume, minimizing the drop in cardiac output through the Frank- Starling mechanism.

Nanoparticles characterization using the CAM assay.

The current chapter highlights the use of chorioallantoic membrane (CAM) of fertilized chicken egg for the characterization of nanoparticles applied in cancer nanomedicine. The CAM assay represents a promising alternative to mouse models in term of costs, ease of use, rapidity and ethical issues in particular for the screening of nanoformulations. Hence, the features of nanoparticles including blood retention, biocompatibility, active targeting or tumor accumulation, angiogenic activity, drug delivery and tumor elimination might be simply evaluated via the CAM model. In particular, in this model, embryo organs and morphology, CAM vasculature and blood cells, transplanted tumors on CAM were typically monitored and used for the evaluation of the nanomaterials. With the above advantages, the CAM assay, as highly valuable in vivo model, could be used regularly in pharmaceutical industries.

New naphthopyran analogues of LY290181 as potential tumor vascular-disrupting agents

A series of 19 analogues of the antiproliferative naphthopyran LY290181 were prepared for structure–activity relationship studies. We found the best activities for test compounds bearing small substituents at the meta position of the phenyl ring. The mode of action of LY290181 and eight new analogues was studied in detail. The compounds were highly anti-proliferative with IC50 values in the sub-nanomolar to triple-digit nanomolar range. The new analogues led to G2/M arrest due to interruption of the microtubule dynamics. In 518A2 melanoma cells they caused a mitotic catastrophe which eventually led to apoptosis. The naphthopyrans also induced a disruption of the vasculature in the chorioallantoic membrane (CAM) of fertilized chicken eggs as well as in xenograft tumors in mice. In a preliminary therapy trial, the difluoro derivative 2b retarded the growth of resistant xenograft tumors in mice.

Effect of pesticides mixture of dimethoate and methidathion on acetylcholinestrase during embryo development using chick embryo model

Pesticides mixture (dimethoate 30% and methidathion 40%) prepared by the farmers in Yemen and engaged in the cultivation of Qat, which is chewed by people every day including pregnant women. Therefore, the developing embryos in the society are more vulnerable than adults to the chronic cholinergic intoxication. This study aimed to examine the chronic effect of pesticides mixture on the AChE of developmental embryo in an avian model, which does not share the maternal potential confounds. For this, fresh fertile chicken eggs (Gallus gallus domesticus) were used. The lethal concentration of pesticides mixture for 50% killing (LD50) values was computed on the basis of probit analysis and was found to be 40 ppm. 1/5 th LD50 and 1/10 th LD50 (8 and 4 ppm) were chosen to be the tested doses. Eggs weighing 54±1 gm were separated in to 3 batches of 10 eggs each batch. One batch of embryos was injected with normal saline and the other batches of embryos injected with pesticides mixture of 4 and 8 ppm each alternative day starting from incubation day 7 for 2 weeks. On day 21 after 12 hours of the last dose an amount of 200 µl blood was collected from the blood vessels surrounding the embryonic membranes and the heamolyzate was used for the assessment of the AChE activity calorimetrically. Result of this study indicated that 1/10 th of the LD50 had only marginal effect on the AChE activity (40.6%). Whereas 1/5 th of the LD50 of pesticides mixture caused significant inhibition of AChE activity (69%) which could not be reversible. So neuro-developmental consequences such as behavioral changes and memory impairment may prolong throughout the life span of the embryo.

Vascular apoptosis associated with meglumine antimoniate: In vivo investigation of a chick embryo model

The vasculo-toxic effect of meglumine antimoniate (MA) was confirmed in our previous investigation. The current study investigates the association of this effect with altered VEGF-A and VEGF-R2 expression. Additional mechanisms by which MA causes vascular toxicity are not clearly understood. We hypothesized that MA may alter normal expression of apoptotic genes and cause vascular toxicity. The current investigation was designed to address this issue using a chick embryo model. Fertile chicken eggs were treated with MA and the extra-embryonic membrane (EEM) vasculature was evaluated by morphometric, molecular and immunohistochemistry assays. The results showed that MA not only altered apoptotic gene expression, but that this alteration may disturb the normal development of the vascular network and cause embryo malformation. The relative expression level of the CASP3, CASP7, CASP9, APAF1, AIF1 and TP53 genes increased in drug-exposed EEMs. In addition, IHC assay confirmed the low expression BCL2 and increased expression of Bax, which are associated with a high rate of apoptosis. We suggest that induction of an apoptotic signaling pathway can lead to vascular defects during embryo development and the consecutive cascade of events can lead to the embryo malformation.

Thermal manipulation during broiler chicken embryogenesis increases basal mRNA levels and alters production dynamics of heat shock proteins 70 and 60 and heat shock factors 3 and 4 during thermal stress.

This study investigated the effects of thermal manipulation (TM) during embryonic days 12 to 18 on the cloacal temperature (Tc) and the kinetics of muscle mRNA levels of heat shock proteins 70 and 60 (Hsp70 and Hsp60) and heat shock factors 3 and 4 (HSF3 and HSF4) during the first week of life and during thermal stress (TS). One thousand five hundred fertile chicken eggs were randomly divided into 5 groups: control group (37.8°C), TM1 (38.5°C for 18 h), TM2 (39°C for 18 h), TM3 (39.5°C for 18 h), and TM4 (40°C for 18 h). On post-hatch days 14 and 28 of age, 30 randomly selected chicks from each group were thermally stressed at 41.0°C for 6 h, while another 30 randomly selected chicks from each group were kept under thermo-neutral conditions. The Tc of TM chicks was only numerically lower than that of the control during the study period. However, during TS at days 14 and 28 of age, the Tc of TM chicks was significantly lower than that of the controls. On post-hatch days 14 and 28, the basal mRNA levels of Hsps and HSFs were significantly higher than those of the control. Furthermore, during TS, rapid increases in the mRNA levels of Hsps and HSFs in the TM groups were observed. These results indicate that, as well as altering their basal mRNA levels during the first week post-hatch, TM also altered the dynamics of the mRNA expression of HSPs and HSFs, which was associated with improved acquisition of thermotolerance during TS.

Evaluation of the toxicopathological lesions of Calotropis procera using a chick embryonic model

Toxicopathological effects of herbs have always been a major concern. There is scant information available about the toxicopathological effects of Calotropis procera (C. procera) in the fetus. Since the chick embryo is a suitable preclinical model to evaluate the toxicopathological effects of chemicals, the objective of this study is to evaluate the lesions of various dosages of C. procera using a chick embryonic model. Fertile chicken eggs were divided into three equal treatment groups; phosphate buffered saline-injected group and C. procera-injected groups whose individuals were treated with C. procera extract at dosages of 50 or 100 mg/kg egg-weight. Embryos were re-incubated post-treatment and allowed to develop until day 18, after which they were examined for macroscopic and microscopic lesions. Although the embryos which were treated with 50 mg/kg egg-weight of C. procera extract macroscopically were normal as well as the controls, those treated with 100 mg/kg egg-weight were stunted and under developed. Microscopic evaluations showed that in embryos treated with 100 mg/kg egg-weight of C. procera, the brain was congested, and severe dilation of central veins and sinusoids as well as hepatocellular degeneration was occurred in liver. Moreover, the kidney and lung were under-developed, but the structure of the heart was normal. Based on our findings, C. procera at dosage of 100 mg/kg is toxic to the chick embryo or/maybe to human fetus during growing period. Further studies are needed to clarify the toxic effects of this plant on the development of fetus.

Expression of Genes Encoding for Xenobiotic Metabolism After Exposure to Dialkylnitrosamines in the Chicken Egg Genotoxicity Alternative Model.

The Chicken Egg Genotoxicity Assay (CEGA) demonstrated responsiveness to various DNA-reactive chemicals requiring metabolic activation, which implies broad bioactivation capability. To assess potential metabolic competence, expression profiles of metabolic genes in the embryo-chicken fetal liver were determined using microarray technology. Fertilized chicken eggs were injected under the CEGA protocol with vehicle (deionized water [DW]), the activation-dependent carcinogens, diethylnitrosamine (DEN), and N-nitrosodiethanolamine (NDELA) at doses producing no effect on survival. Previously in CEGA, DEN produced DNA damage, whereas NDELA did not. Expressions of 463 genes known to encode for phase I and II of endo- and xenobiotic metabolism were detected on the array. DW did not affect the expression of the selected genes, deregulating less than 1% of them. In contrast, DEN at 2 mg/egg and NDELA at 4 mg/egg produced significant transcriptomic alterations, up-regulating up to 41% and down-regulating over 31% of studied genes. Both nitrosamines modulated the majority of the genes in a similar manner, sharing 64 up-regulated and 93 down-regulated genes with respect to control group, indicating similarity in the regulation of their metabolism by avian liver. Differences in gene expression between DEN and NDELA were documented for several phase I CYP 450 genes that are responsible for nitrosamine biotransformation, as well as for phase II genes that regulate detoxication reactions. These findings could underlie the difference in genotoxicity of DEN and NDELA in CEGA. In conclusion, the analysis of gene expression profiles in embryo-chicken fetal liver dosed with dialkylnitrosamines demonstrated that avian species possess a complex array of inducible genes coding for biotransformation.

Evaluation the Toxicopathological Lesions of Berberis Vulgaris Using a Chicken Embryonic Model

Toxicopathological effects of herbs have always been a major concern. There is scant information available about the toxicopathological effects of barberry in the fetus. Since the embryogenesis in chicken is similar to human beings, the objective of this study is to evaluate the lesions of the various dosages of Berberis vulgaris using a chicken embryonic model. Fertile chicken eggs were divided into four equal treatment groups; phosphate buffered saline-injected group and barberry-injected groups whose individuals were injected with Berberis vulgaris fruit-extract at dosages of 10, 50 and 100 mg per Kg egg-weight, respectively. Embryos were re-incubated post-treatment and allowed to develop until day 18, after which they were examined for macroscopic and microscopic lesions. Results showed that embryos were stunted in the barberry-injected groups. Defect in feather growth and general congestion was accompanied by pathological changes in brain, liver, kidney, heart and lung. Histopathological lesions include congestion, hemorrhage, edema and micro-thrombosis in the affected organs. Based on findings, it is concluded that Berberis vulgaris at the above-mentioned concentrations is toxic to the chicken embryo in a dose dependent manner. Further studies are needed to clarify the toxic effects of this herb on the development of human fetus.

ESTIMATION OF CADMIUM FROM DRINKING WATER SOURCES AND ITS EFFECT ON CHICK EMBRYO DEVELOPMENT

The aim of the study was to screen Cd contamination from the River Mutha and to investigate its effects on chick embryo development. The study was correlated with the toxicity of fetal development in humans, as chick embryo provides an excellent model system for studying the development of higher vertebrates including humans. The estimated average concentration of Cd in water column of River Mutha was 0.0986 mg/l, exceeded the safe limit of drinking water (0.0003 mg/l). To study the effects of Cd on chick embryo development, analogous concentration observed in River water was introduced in fertilized chicken eggs. Aqueous solutions of Cd ranging from 0.05 to 1.00 mg/l were injected into fertilized chicken eggs at 72 hours of incubation and metal injected eggs were further incubated for 10 days to see the toxic effects if any. Control eggs were treated in the similar way with sterile distilled water. Results showed abnormal development of blood vessels and decrease in the embryo survival rate in the Cd treated eggs.

Quantitative Analysis of the Area of the Apical Ectodermal Ridge in Chick Appendages Using Image-J.

To determine the effect of sodium phenytoin on the apical ectodermal ridges (AER) of chick wing buds by using the software program Image-J. An experimental study. Department of Anatomy, Regional Center, College of Physicians and Surgeons Pakistan (CPSP), Islamabad, from January 2014 to January 2015. Sixty fertilised chicken eggs of 'Egyptian fayoumi' breed were selected and separated into experimental (B) and control (A) groups, each having 30 eggs. A single dose of 3.5 mg sodium phenytoin was injected into each egg of the experimental group. The controls were injected with the same volume of normal saline. Developing embryos were extracted 96 hours (day 4) after incubation and histological sections were cut at 5 μm thickness. These sections were stained with Feulgen Nuclear and Light Green. The area of apical ectodermal ridges of chick wing buds was calculated by employing Image-J and subjected to statistical analysis. The difference between the mean values of the area of apical ectodermal ridges of experimental and control groups, as calculated by Image-J, was found to be statistically insignificant. Change in the area of the apical ectodermal ridges in experimental chicks, following phenytoin exposure, was insignificant as proven on the basis of quantification by Image-J.

New (arene)ruthenium(II) complexes of 4‑aryl‑4H‑naphthopyrans with anticancer and anti-vascular activities

A series of four 2‑amino‑3‑cyano‑4‑(3/4‑pyridyl)‑4H‑benzo[h]chromenes 2a–d and their dichlorido(p‑cymene)ruthenium(II) complexes 3a–d were tested for antiproliferative, vascular-disruptive, anti-angiogenic and DNA-binding activity. The coordination of the 4‑pyridyl‑4H‑naphthopyrans 2 to ruthenium led to complexes with pleiotropic effects. Unlike the free ligands 2a–d, their ruthenium complexes 3a–d showed a significant affinity for DNA as demonstrated by electrophoretic mobility shift assays (EMSA) and ethidium bromide assays. Binding of 3a–d to calf thymus DNA proceeded about 10-times faster compared with cisplatin. Treatment of HT-29 colon carcinoma, 518A2 melanoma and MCF-7Topo breast cancer cells with 3a and 3b caused an accumulation of cells in the G2/M phase and an increase of the fraction of mitotic cells in the case of HT-29, due to alterations of the microtubule cytoskeleton as shown by immunofluorescence staining. Complexes 3b–c showed a dual effect on the vascular system. They suppressed angiogenesis in zebrafish embryos and they destroyed the vasculature of the chorioallantoic membrane (CAM) in fertilized chicken eggs. They also inhibited the vasculogenic mimicry, typical of U-87 glioblastoma cells in tube formation assays.

Thermal Manipulation During Brolier Chicken Embryogenesis Increases the Basal mRNA levels of antioxidant factors and Alters Their Production Dynamics during Thermal Stress

This study investigated the effects of thermal manipulation during embryonic days 12–18 on the kinetics of the liver mRNA levels of antioxidant factors (Catalase, Glutathione (GSH), Superoxide Dismutases (SOD)) during thermal stress. One thousand fertile chicken eggs were randomly divided into four groups: Hubbard Control group (37.8°C), Hubbard TM (39°C for 18 h), Cobb Control group (37.8°C), Cobb TM (39°C for 18 h). On post‐hatch days 14 and 28 of age, 30 randomly selected chicks from each group were thermally stressed at 41.0°C for 6 h while another 30 randomly selected chicks from each group were kept under thermo‐neutral conditions. The Tc of TM chicks was only numerically lower than that of the control during the study period. However, during TS at days 14 and 28 of age, the Tc of TM chicks was significantly lower than that of the control. On post‐hatch days 14 and 28, the basal mRNA levels of antioxidant factors were significantly higher than those of the control. Furthermore, during TS, rapid increases in the mRNA levels of antioxidant factors in the TM groups were observed. These results indicate that, as well as altering the basal mRNA levels of antioxidant factors during the first week post‐hatch, TM also altered the dynamics of the mRNA expression of antioxidant factors, which was associated with improved acquisition of thermotolerance during thermal stress.Support or Funding InformationDeanship of research/Jordan University of Science and Technology (Project#: 125/2017)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

The Chick Chorioallantoic Membrane (CAM) as a Multi‐Purpose Preclinical Model in Oncology

The chorioallantoic membrane (CAM) of fertilized chick eggs is a preclinical in vivo model and an alternative or pretest for animal studies in rodents. Although the CAM xenotransplantation model is a reproducible, reliable and efficient model conforming to the 3R principle to reduce mammalian animal experiments, and the United States Food and Drug Administration recommended the CAM for in vivo studies on angiogenesis, the method is not very common so far. Undoubtedly, the CAM assay has its limitations: The experimental window of about 10 days is a limiting factor hampering the establishment of the model in oncology. Here we show the implementation of the CAM model in oncological research on modern diagnostics and therapeutics. Tumor cells were xenotransplanted onto the chorioallantoic membrane, which serves as nutritious natural substrate for their growth and allows the three‐dimensional formation of solid tumors in an in vivo microenvironment. We successfully transplanted cancer cells stably expressing firefly luciferase, e.g. breast and lung cancer cells, and established methods for direct in ovo monitoring after the topical or intravenous administration of diagnostics or therapeutics. Upon administration of doxorubicin, a commonly used chemotherapeutic, the growth of cancer xenografts on the CAM was monitored by bioluminescence imaging using an IVIS in vivo Imaging System after the application of D‐luciferin for up to 3 days after treatment. The obtained results were quantified and validated by immunohistochemical analysis of proliferation and apoptosis (Ki‐67, Tdt) upon tumor tissue extraction. After labelling of compounds with gadolinium (Gd) as a contrast agent, high‐resolution magnetic resonance (MR) imaging at several time‐points enabled the evaluation of the compound's bio‐distribution and accumulation in tumor tissue. To confirm the MR findings, organs were extracted and analyzed for the Gd contents by inductively coupled plasma optical emission spectrometry (ICP‐OES).These methods facilitate the straightforward and non‐invasive assessment of a compound's therapeutic efficacy and biodistribution and therefore possess implications for the development of new pharmacological substances.The studies were conducted in conformance with the FASEB Statement of Principles for the use of Animals in Research and Education.Support or Funding InformationThis work was supported by the Volkswagen Foundation.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Antiangiogenic evaluation of ZnWO4 nanoparticles synthesised through microwave-assisted hydrothermal method

Angiogenesis, the complex process of formation of new blood vessels from pre-existing blood vessels, which involves the participation of several pro- and anti-angiogenic factors, is implicated in many physiological and pathological conditions. Nanoparticle-based anti-angiogenic activity at the tumour tissue, harnessed by the Enhanced Permeability and Retention Effect (EPR effect), could potentially become a breakthrough therapy to halt tumour progression. Herein, we evaluate the anti-angiogenic effect of ZnWO4 nanoparticles (NPs). The nanoparticles were obtained by microwave-assisted hydrothermal synthesis (MAHS) at 120 °C for 60 min and were structurally characterised by X-ray diffraction (XRD) and micro-Raman (MR) spectroscopy. The mean size and polydispersity index were estimated by Zeta potential analysis. The XRD analysis revealed structural organisation at a long-range order, with an average crystallite size of around 3.67 nm, while MR revealed short-range order for ZnWO4. The anti-angiogenic potential of zinc tungstate nanoparticles was investigated through the chorioallantoic membrane assay (CAM) using fertilised chicken eggs. We demonstrate, in an unprecedented way, that nanocrystalline ZnWO4 NPs obtained by MAHS, at low reaction temperatures, showed excellent anti-angiogenic properties even at low concentrations. The ZnWO4 NPs were further evaluated for its cytotoxicity in vitro.

Non-thermal microwave effects on chicken foetuses during organogenesis

The purpose of this work is το investigate possible effects of pulse-modulated (PW) and non-modulated (CW) low power density microwave radiation on chicken foetuses during organogenesis. A total of 496 fertilised chicken eggs were used in this study; 380 of diem as experimental material and 116 as controls. The experimental eggs were exposed to a non-thermal power density of 8.8 uW/cm2 (SAR= 1.16 mW/Kg) radiation at 9.152 GHz, from the 3rd to the 10th day of incubation, continuously. 172 eggs were irradiated by CW whereas 208 eggs by PW. The controls were sham-exposed. The foetuses of the experimental eggs and of the controls were macro- and microscopically examined at the end of their incubation. In 62.78% of CW-irradiated, 47.12% of PWirradiated and only in 3.44% of the sham-exposed fetuses abnormalities (development retardation and severe malformations) and embryonic and foetal deaths (embryonic and foetal) were observed. The differences in the effects caused by CW and PW irradiation were statistically evaluated. These results support the aspect that very low power density microwaves applied to the chicken foetuses, in ovo during organogenesis, may cause abnormal development.

Presentation of a variation of the chorioallantoic membrane set up as a potential model for individual therapy for squamous cell carcinoma of the oropharynx.

The chorioallantoic membrane of fertilized chicken eggs in an early phase of breeding presents an approved test situation for the growth and treatment of human cancer cells.These models work due to the inoculation of cells into the membrane that stays within the egg shell during the time of investigation. In this study a modification of this model is presented. Samples of native tumors, rather than cell lines, are transplanted into the membrane and the body of the egg is taken out of the shell and placed in a plastic bowl. These modifications lead to an enhanced accessibility to the chorioallantoic membrane and the surrounding vessels thus facilitating intra venous access and application of pharmaceuticals and a focused radiotherapy. With the current modifications the embryo was kept alive and additionally, the vascularized tumor environment was preserved.