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Purpose: Cancer stem cells (CSCs) reported in various tumors play a crucial role in tumorigenesis and metastasis of retinoblastoma (Rb). Following the efforts to reduce, replace, and refine the use of mammalian models, we aimed to establish a short?term xenograft for Rb to evaluate the CSC properties of CD133? Rb Y79 cells, using the well?established chick embryo chorioallantoic membrane (CE?CAM) assay. Methods: Y79 cells were cultured, labeled with two different dyes (CM?Dil Y79 and enhanced green fluorescent protein (eGFP)) and sorted for CD133? and CD133 + subsets. Two million cells from each of the labeled groups were transplanted onto the abraded CAM on embryonic day 7 (E7). On E14, the tumor nodule formation on CAM and spontaneous metastasis to the embryos were evaluated by confocal microscopy, in vivo imaging, and histology. Results: Y79 cells formed pink–white raised perivascular nodules with feeder vessels on the CAM with both the types of labeled CD133? cells. CD133? cells, when compared to CD133 + cells, demonstrated significantly larger tumor volume (40.45 ± 7.744 mm3 vs 3.478 ± 0.69 mm3, P = 0.0014) and higher fluorescence intensity (CM?Dil: AUF = 6.37 × 107 ± 7.7 × 106 vs 1.08 × 107 ± 1.6 × 106; P < 0.0001; eGFP: AUF = 13.94 × 104 ± 2.54 × 104 vs AUF = 1.39 × 104 ± 0.4 × 104; P = 0.0003). The metastatic potential of CD133? cells was also observed to be higher as noted by in vivo imaging and histopathology. Conclusion: This study highlights that CE?CAM is a feasible alternative nonmammalian model for evaluating tumorigenicity and metastatic potential of Y79 CSCs. Increased tumorigenicity and metastatic potential of CD133? subset of tumor cells substantiate their CSC properties
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The Hancornia speciosa latex has shown angiogenic activity. Angiogenesis plays a major role in wound healing, and materials that stimulate this process could be used to develop drugs. This study aimed to explain the role of proteins in the H. speciosa serum fraction latex in angiogenesis. Hence, this material was treated with proteinase K and the proteins were inactivated. After protein inactivation, angiogenic activity was assessed with the chick chorioallantoic membrane assay. The result showed that the proteins in the serum fraction are responsible for angiogenic activity. Then, the total protein content in the serum fraction and its enzymatic activity were investigated. The low protein content observed in the H. speciosa serum fraction latex suggests that this biomaterial could be used to develop new drugs with a hypoallergenic response. Despite the low protein content, there was a significant enzymatic activity of at least three enzymes in the serum fraction latex: ß-1,3 glucanase, ß-glucosidase, and proteases. These enzymes seem to influence the healing process, assisting debridement, extracellular matrix remodeling, and collagen deposition, and decreasing the chances of contamination by microorganisms. In conclusion, the enzymes in the H. speciosa serum latex are associated with the angiogenic activity of this biomaterial and may be used to assist the wound healing process.
Subject(s)
Wound Healing , Endopeptidase K , Apocynaceae , Enzymes , LatexABSTRACT
[Abstract] Objective To investigate the effects of the downregulation of draxin expression on the projection characteristics of 23C10-positive neural fibers in the chick embryonic hindbrain. Methods The vitro incubation of HH stages 21-22 chick embryonic hindbrain biopsy with alkaline phosphatase (ALP) protein was used as control group. The incubation of HH stages 21-22 chick embryonic hindbrain biopsy with draxin-ALP fusion protein was used as experimental group. The number of embryonic hindbrain for each group was 10. To detect whether 23C10-positive neural fibers could directly bind to draxin protein or not;In ovo electroporation using empty vector in the chick embryonic hindbrain was used as control group. In ovo electroporation with small interfering RNA(siRNA) expressing vector for reducing draxin expression in the chick embryonic hindbrain was used as experimental group. The number of embryonic hindbrain for each group was 18. The effect of the down-regulation of draxin expression and the change of projection characteristics of 23C10-positive neural fibers were observed to check whether the down-regulation of draxin expression would affect the distribution of 23C10-positive fibers. Results Most portion of draxin protein could overlap with 23C10-positive neural fibers in HH stages 21-22 chick embryonic hindbrain biopsies; After expression of the siRNA plasmid against draxin by electroporation, the expression level of draxin protein was significantly reduced, and the distribution of 23C10-positive fibers was scattered in the dorsal hindbrain on the electroporated side at HH stages 25-26 of chick embryos (P < 0. 05) . Conclusion Draxin protein may directly bind to 23C10-positive fibers in hindbrain, and it plays an important regulatory role in the fasciculation of 23C10-positive fibers during chick embryonic development.
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Abstract Introduction: Rhinella schneideri is a toad widely distributed in South America and its poison is characterized by inducing cardiotoxicity and neurotoxicity. Objective: In this work, we investigated pharmacological strategies to attenuate the peripheral neurotoxicity induced by R. schneideri poison in avian neuromuscular preparation. Methods: The experiments were carried out using isolated chick biventer cervicis preparation subjected to field stimulation for muscle twitches recordings or exposed to acetylcholine and potassium chloride for contracture responses. Results: Poison (10 μg/ml) produced complete neuromuscular blockade in chick biventer cervicis preparation within approximately 70 min incubation (times for 50 and 90 % blockade: 15 ± 3 min and 40 ± 2 min, respectively; P < 0.05, N= 5); contracture responses to exogenous acetylcholine and KCl were unaffected by poison indicating no specificity with postsynaptic receptors or myotoxicity, respectively. Poison (10 μg/ml)-induced neuromuscular blockade was not prevented by heparin (5 and 150 IU/ml) under pre- or post-treatment conditions. Incubation at low temperature (23-25 °C) abolished the neuromuscular blockade; after raising the temperature to 37 °C, the complete neuromuscular blockade was slightly slower than that seen in preparations directly incubated at 37 °C (times for 50 and 90 % blockade: 23 ± 2 min and 60 ± 2.5 min, respectively; P < 0.05, N= 4). Neostigmine (3.3 μM) did not reverse the neuromuscular blockade in BC preparation whereas 3,4-diaminopyridine (91.6 μM) produced a partial and sustained reversal of the twitch responses (29 ± 7.8 % of maximal reversal reached in approximately 40 min incubation; P < 0.05, N= 4). Conclusions: R. schneideri poison induces potent peripheral neurotoxicity in vitro which can be partially reversible by 3,4-diaminopyridine.
Resumen Introducción: Rhinella schneideri está ampliamente distribuida en Suramérica y su veneno es caracterizado por inducir cardiotoxicidad y neurotoxicidad. Objetivo: En este trabajo, investigamos estrategias farmacológicas para atenuar la neurotoxicidad periférica inducida por el veneno de R. schneideri en preparaciones neuromusculares de aves. Métodos: Los experimentos fueron realizados usando preparaciones de biventer cervicis de pollos sometidas a estimulación de campo para el registro de las contracciones musculares o expuestas a la acetilcolina y al cloruro de potasio para la respuesta contractural. Resultados: El veneno (10 µg/ml) provocó un bloqueo neuromuscular completo en las preparaciones después de aproximadamente 70 min de incubación (tiempos para 50 y 90 % de bloqueo: 15 ± 3 min y 40 ± 2 min, respectivamente; P < 0.05, N = 5); las contracturas en respuesta a la acetilcolina y el KCl exógenos no fueron afectadas por el veneno, indicando que no hay una interacción especifica con receptores postsinápticos o miotoxicidad respectivamente. El bloqueo neuromuscular causado por el veneno (10 µg/ml) no fue prevenido por la heparina (5 y 150 UI/ml) bajo condiciones pre y post-tratamiento. La incubación a bajas temperaturas (23-25 ºC) abolió el bloqueo neuromuscular; después de aumentar la temperatura a 37 ºC, el bloqueo neuromuscular total fue levemente más lento que el visto en preparaciones directamente incubadas a 37 ºC (tiempos para 50 y 90 % de bloqueo: 23 ± 2 min y 60 ± 2.5 min, respectivamente; P < 0.05, N= 4). Neostigmina (3.3 µM) no revirtió el bloqueo neuromuscular, mientras que 3.4-diaminopiridina (91.6 µM) produjo una reversión parcial y sostenida de las respuestas neuromusculares (29 ± 7.8 % de la reversión máxima alcanzada en aproximadamente 40 min de incubación; P < 0.05, N = 4). Conclusiones: El veneno de R. schneideri indujo neurotoxicidad periférica potente in vitro, el cual puede ser revertido por 3.4-diaminopiridina.
Subject(s)
Animals , Bufo marinus , Neuromuscular Blockade , Birds , BrazilABSTRACT
Mitochondrial shape rapidly changes by dynamic balance of fusion and fission to adjust to constantly changing energy demands of cancer cells. Mitochondrial dynamics balance is exactly regulated by molecular motor consisted of myosin and actin cytoskeleton proteins. Thus, targeting myosin-actin molecular motor is considered as a promising strategy for anti-cancer. In this study, we performed a proof-of-concept study with a natural-derived small-molecule J13 to test the feasibility of anti-cancer therapeutics
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Background & objectives: Cataract is one of the leading causes of blindness in the world. The aim of the present study was to investigate anticataractogenic effect of betaine in chick embryo hydrocortisone (HC)-induced cataract model. Methods: The study included 60 fertilized eggs divided into six groups each having 10 eggs: one group treated with only HC (HC group); three treated with both HC and different doses of betaine (HC/B 1.00, HC/B 0.50 and HC/B 0.25 groups) and two non-HC groups treated with only phosphate-buffered saline (PBS group) or betaine (B group). After the injections, lenses of the embryos were removed and classified into five stages according to the lens opacification. The amounts of reduced glutathione (GSH) in the removed lenses were measured. Results: All the lenses in non-HC-treated groups were clear, whereas in the HC-treated group, 90 per cent of the lenses had cataract (stages 4 and 5). The mean score of lens opacity was significantly lower in all HC/B groups compared to HC group (2.4-3.5 vs. 4.4, P<0.05). Among HC/B groups, the HC/B 0.25 group had significantly lower mean score of lens opacity compared to remaining HC/B groups treated with higher doses of betaine. In addition, the mean reduced GSH level was significantly higher in HC/B 0.25 group compared to HC, HC/B 1.00 and HC/B 0.50 groups (P<0.001). Interpretation & conclusions: The present results show beneficial anti-cataract and anti-oxidant effects of 0.25 ?mol/egg betaine on HC-induced cataract in the chick embryo.
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Introduction: Evaluating the patients who presents with feverand rash can be challenging because the differential diagnosisis extensive and includes minor and life threatening illnesses.Febrile rashes are classified into maculopapular rash,generalized diffuse erythema, and vesicular. Currently, viralexanthems are by far the most common cause of fever withrash in children. Study aimed to identify the patients presentingwith febrile rash and to confirm cases by laboratory diagnosticmethods to ensure fast, economical and speedy confirmationof the cause.Material and methods: A survey was carried out fromMay 2017 to Sep 2018 by the Department of Microbiologywith technical support from OP and IP staff. Blood sampleswere collected after informed consent. Serum samples werescreened for immunoglobulin M by ELISA, test was processedaccording to the manufacturer’s instructions.Results: This study covered 215 subjects out of which (55.9%)were males and (43.6%) were females, cases were segregatedwith different age groups 0 – 5 years, 6 – 12years, 13-18 yearsand adults > 18 years who developed febrile rash with fever.All the samples were screened for virus specific IgM.Conclusion: There is a need for routine surveillance programat every teaching Hospital setting to ensure the incidenceof the febrile rash with illness in children among differentage groups. Diagnosis can be established serologically byemphasizing the need for Clinico-serological correlationof rashes. As antibiotics will be required to treat bacterialdiseases, knowing the viral etiology will help in avoidingunnecessary administration of antibiotics.
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Background: Swertia chirata has been an important herb known for centuries for its various medicinal uses and bitter taste. The stem of the plant is used as a traditional medicine in an array of diseases including the treatment of vomiting. Therefore, the study was undertaken to explore the possible antiemetic property of methanolic extract of its stems by using chick emesis model.Methods: 25 male chicks of four days old weighing 25 to 35 grams were fed with copper sulfate anhydride at 50mg/kg body weight to induce emesis. The chicks were grouped into 5 with each group bearing 5 chicks (n=5). Group I (control) received 10ml/kg normal saline; group II (standard) received 150mg/kg chlorpromazine; group III (experimental-1), group IV (experimental-2) and group V (experimental-3) received 50, 100 and 150mg/kg respectively of the extract. All doses are given intraperitoneally. Assessment of antiemetic activity was done by calculating the percentage of inhibition of the number of retches in the chicks.Results: All the three doses of the extract showed antiemetic activity. The dose of 50 mg/kg showed activity comparable to chlorpromazine, while dose of 100 mg/kg and 150mg/kg showed greater activity than chlorpromazine. Highest antiemetic activity (79.26% inhibition) was shown by a dose of 150mg/kg and lowest (42.22% inhibition) by 50mg/kg.Conclusions: Methanolic extract from the stems of Swertia chirata has excellent anti-emetic property which can be further investigated for development of potential antiemetic medicines.
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Purpose: The aim of this study was to present an experimental optical coherence tomography (OCT)–guided anterior segment (AS) imaging chick embryo model. Through this model, we aimed to reveal similarities and differences between human cornea, AS tissues, and chick embryo tissues by quantitative image analysis. Methods: Ex vivo, the chick embryos' globes were determined by detailed AS camera of spectral-domain (SD)-OCT in 10 fertilized specific pathogen-free eggs on the 20th day. Quantitative image analysis of anterior chamber tissues was performed with SD-OCT in detail. After imaging, cross sections of the chick embryo globes containing cornea with anterior chamber were histologically examined and compared with human tissues. The similarities of our model with data in the human cornea and AS studies in the literature were compared. Results: SD-OCT imaging was able to successfully delineate the AS tissues of chick embryos such as the cornea, iris, lens, pupil, conjunctiva, ciliary body, anterior chamber, and lens. Quantitative semi-automated measurements showed the following: mean central corneal thickness: 213.4 ± 7.05 ?m (197–223 ?m), mean anterior chamber depth: 878.9 ± 41.74 (804–919 ?m), mean anterior chamber area: 2.43 ± 0.16 mm2 (2.17–2.73 mm2), mean corneoscleral junction (limbal) thickness: 322.8 ± 20.05 ?m (289–360 ?m), and mean iris thickness: 230.4 ± 13.27 ?m (203–245 ?m). In addition, detailed histological comparisons of the AS tissues with human tissues were evaluated to be very similar. Conclusion: In conclusion, this chick embryo model mimics human tissues and it can be considered as a platform for the study of teratogen-induced malformations and AS dysgenesis during gestation of AS tissues. In addition, this study demonstrates the feasibility of SD-OCT in the quantitative assessment of AS structures in chick embryo model.
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Background: Angiogenesis is important for the typical physiological activities such as cure from injury, menstrual cycle and embryo growth. It is also plays a crucial role in several pathological conditions in cancer. Antiangiogenesis, e.g., inhibition of blood vessel growth, is being investigated as a way to prevent the growth of tumors and other angiogenesis-dependent diseases. The chick embryo chorioallantoic membrane (CAM) is commonly used as an experimental in vivo assay to study both angiogenesis and antiangiogenesis in response to tissues, cells or soluble factors. Given the high occurrence of cancer worldwide and the major source of the discovery of new lead molecules are medicinal plants. The objective of the present research was to study the antiangiogenic property of “aqueous extract of Nigella sativa seeds” using chick chorioallantoic membrane (CAM) assayMethods: The chick chorioallantoic membrane (CAM) assay for screening the effect of Nigella sativa on anti-angiogenesis was performed according to the method given by Ribatti and co-workers.Results: The results of present study significantly increased the antiangiogenic effect on CAM by decreasing the proliferation of capillary networks in a dose (50 to 300 µg/egg) dependent manner which is probably related to the inhibition of neovascularization.Conclusions: It is concluded that aqueous extract of N. sativa seeds possesses significant antiangiogenic activity, and this is a possible rationale for its folkloric use as an anticancer agent.
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In order to compare Wnt/ beta-catenin expression in mouse and chick facial primordia during development, we performed an immunohistochemical analysis of both protein expressions in E12 to E17 mouse embryos and in E3 and E10 chick embryos. During odontogenesis, from bud to bell stage, both proteins exhibit similar fixation patterns, with epithelial and mesenchymal immunoreactivity, consistant with literature data. Double labelling demonstrates that the same cells express both antigens, even in undifferentiated mesenchyme. The enamel knot, and the ameloblastic and odontoblastic layers are stained at the same manner. In the chick, Wnt and beta-catenin are diffusely present on craniofacial mesenchyme. In both species, premuscular blastemata express Wnt and b-catenin, but Wnt is specifically expressed on the perichondrium and ossification centers, suggesting a role independent from beta-catenin pathway.
Subject(s)
Animals , Chick Embryo , Mice , Tooth/embryology , Embryo, Nonmammalian/metabolism , Wnt Proteins/metabolism , beta Catenin/metabolism , Immunohistochemistry , Fluorescent Antibody TechniqueABSTRACT
ABSTRACT The phytochemical study of Galium tunetanum Lam., Rubiaceae, leaves led to the isolation of 13 compounds from the chloroform-methanol and the methanol extracts, including six iridoid glycosides, one non-glycoside iridoid, two p-coumaroyl iridoid glycosides, two phenolic acids, and two flavonoid glycosides. The structural determination of the isolated compounds was performed by mono- and bidimensional NMR spectroscopic data, as well as ESI-MS experiments. All compounds were isolated from this species for the first time. The anti-angiogenic effects of the isolated iridoids were also reported on new blood vessels formation using the chick embryo chorioallantoic membrane as in vivo model. Results showed that among the isolated iridoids tested at the dose of 2 µg/egg, asperuloside (1), geniposidic acid (2), and iridoid V1 (3) reduced microvessel formation of the chorioallantoic membrane on morphological observations using a stereomicroscope. The anti-angiogenic effects of the active compounds, expressed as percentages of inhibition versus control, were 67% (1), 59% (2), and 54% (3), respectively. In addition, the active compounds were able to inhibit angiogenesis in the chorioallantoic membrane assay, in a dose-dependent manner (0.5-2 µg/egg) as compared to the standard retinoic acid.
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Este estudo objetivou avaliar os efeitos da inoculação de proteína isolada de soja (P.I.S.) em ovos embrionados de matrizes semipesadas com diferentes idades. Foram utilizados 320 ovos embrionados de matrizes semipesadas da linhagem Rhode Island Red com 35 e 70 semanas de idade. Ao 17º dia de incubação, os ovos foram inoculados com 0,5mL de solução diretamente na cavidade alantoide. O delineamento experimental utilizado foi o inteiramente ao acaso, em esquema fatorial (2 x 4), constituído por dois fatores: idades das matrizes (matrizes jovens e matrizes velhas) e soluções inoculadas (ovo íntegro; 0,5% de solução salina; 1% de P.I.S. + 0,5% de solução salina; e 2% de P.I.S. + 0,5% de solução salina), totalizando oito tratamentos contendo 40 ovos cada. Os dados coletados foram analisados pelo teste de Tukey a 5% de significância. A inoculação de proteína isolada de soja apresentou efeito positivo sobre a relação pinto-ovo, porém sem proporcionar melhora nos rendimentos de incubação, no desenvolvimento dos órgãos e no desempenho de pintos na fase pré-inicial. A idade da matriz influenciou diretamente as variáveis avaliadas, em que ovos oriundos de aves velhas apresentaram maior peso, maior mortalidade embrionária na fase tardia, pintos mais pesados e pior conversão alimentar.(AU)
This study aimed to evaluate the influence of inoculation of isolated soy protein (P.I.S.) in embryonated eggs of semi-heavy with different ages. 320 embryonated eggs were used from semi-heavy breeders of Rhode Island Red lineage with 35 and 70 weeks of age. In the 17th day of incubation, the eggs were inoculated with 0.5 ml of solution in the allantoic cavity. The experimental method was completely randomized in factorial scheme (2 x 4), consisting of two factors: matrices ages (young and old matrices) and inoculation solutions (non-inoculated eggs; 0.5% of saline solution; 1% of P.I.S. + 0.5% of saline solution; and 2% of P.I.S. + 0.5% of saline solution, totalling eight treatments containing 40 eggs each. The inoculation of isolated soy protein showed positive effect on the chick-egg relation, albeit without improving incubation yields, organ development and chicks's performance in the pre-initial phase. The age of the breeder directly influenced the variables evaluated, with eggs from older breeder showing better weight, higher embryonic mortality in late phase, heavier chicks and worse feed conversion.(AU)
Subject(s)
Animals , Chick Embryo/growth & development , Chickens , Eggs/analysis , Embryonic Development , Proteins/analysis , Glycine maxABSTRACT
The aim of this study was to observe morphological changes of the cultured otocysts isolated from various stages of the chick embryo. Isolated otocysts were dissected from embryonic day, E2.5-4.5 of incubation (HH stage 16-26) according to stages of developing inner ear. Morphology of the chick otocyst exhibited an ovoid shape. The width and height of the otocyst were 0.2 mm and 0.3 mm, respectively. Elongation of a tube-like structure, the endolymphatic duct, was found at the dorsal aspect of the otocyst. The cultured otocyst is lined by the otic epithelium and surrounding periotic mesenchymal cells started to migrate outwards the lateral aspect of such epithelium. Notably, the acoustic-vestibular ganglion (AVG) was observed at the ventrolateral aspect of the otocyst. Appearance of AVG in vitro can be applied for studying chemical-induced ototoxicity and sensorineural hearing loss. It was concluded that the organ-cultured otocyst of the chick embryo could be used as a model to study sensory organ development of avian inner ear.
El objetivo de este estudio fue observar los cambios morfológicos de otocistos cultivados aislados en las diversas etapas del desarrollo del embrión de pollo. Otocistos aislados fueron obtenidos de embriones día, E2.5-4.5 de incubación (HH etapa 16-26) de acuerdo a las etapas de desarrollo del oído interno. El otocisto de pollo presentó una morfología ovoide. El ancho y la altura del otocisto fue de 0,2 mm y 0,3 mm, respectivamente. En la cara dorsal del otocisto se visualizó el alargamiento de una estructura similar a un tubo, el conducto endolinfático. El otocisto cultivado está revestido por epitelio ótico y células mesenquimatosas perióticas que comienzan a migrar hacia el exterior de la cara lateral en búsqueda del epitelio. En particular, el ganglio acústico-vestibular (GAV) fue observado en la parte ventrolateral del otocisto. La aparición de GAV in vitro puede ser aplicado para el estudio de la ototoxicidad inducida por productos químicos y la pérdida de audición neurosensorial. Se concluyó que el otocisto cultivado de embrión de pollo podría ser utilizado como un modelo para estudiar el desarrollo de órganos sensoriales del oído interno aviar.
Subject(s)
Animals , Chick Embryo/anatomy & histology , Ear, Inner/embryology , MorphogenesisABSTRACT
Objective Angiogenesis is the development of new blood vessels. The ion channels on endothelium play a vital action in cell proliferation and so in the related angiogenesis. We aimed to investigate the anti-angiogenic effects of Mefloquine (Cl− channel blocker) and 4-Aminopyridine (K+ channel blocker). Methods The anti-angiogenic activities of Mefloquine and 4-Aminopyridine (4-AP) were investigated by in-vivo (sponge implantation method), in-vitro (aortic ring assay) and in-ovo (CAM, Chick Chorioallantoic membrane) methods. The standard antiangiogenic drug used was Bevacizumab. Results In the CAM assay, both the ion channel blockers exhibited noticeable antiangiogenic activity at the concentrations of 10
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Objective To compare the growth of three different cancer cell lines on chick chorioallantoic membrane (CAM), to select the best transplanted cancer cell line for establishing a transplanted tumor model and to observe the biological characteristics.Methods The human lung cancer cell line A549, human tongue cancer cell line TCA8113 and human liver cancer cell line QGY7703 were respectively inoculated into CAM at the 7th day of age.The chick embryo survival rate, tumor survival rate, tumor formation rate and induced angiogenesis were detected and the growth characteristics of the transplanted tumor model were observed.Results Compared with the groups inoculated with A549 cells and QGY7703 cells, the tumor formation rate of TCA8113 cells was the highest (P < 0.05), to be the best cancer cell line for transplanted tumor.The optimal inoculated number of cells was 8.0×106/chick embryo, the optimal growth period of the tumor was 4~8 d, and the best experiment time was 7 d after inoculation.Conclusion The TCA-CAM transplanted tumor model of tongue squamous cell cancer is successfully established for further study of the biological characteristics and mechanisms of tumor growth, angiogenesis, invasion and metastasis, and provide a good experimental animal model for anti-tumor drug screening.
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Objective: Angiogenesis is the development of new blood vessels. The ion channels on endothelium play a vital action in cell proliferation and so in the related angiogenesis. We aimed to investigate the anti-angiogenic effects of Mefloquine (Cl?channel blocker) and 4-Aminopyridine (K+channel blocker). Methods: The anti-angiogenic activities of Mefloquine and 4-Aminopyridine (4-AP) were investigated by in-vivo (sponge implantation method), in-vitro (aortic ring assay) and in-ovo (CAM, Chick Chorioallantoic membrane) methods. The standard anti-angiogenic drug used was Bevacizumab. Results: In the CAM assay, both the ion channel blockers exhibited noticeable anti-angiogenic activity at the concentrations of 10?5 M and 10?4 M where they significantly exhibited ant proliferative activity by inhibiting the new blood vessel formation. For the further confirmation anti-angiogenic activity was evaluated in vitro and in vivo. In Rat aortic ring assay reduction in the area of sprouts were observed with 40μM of 4-AP and 7 μM of Mefloquine. A significant reduction in weight of sponges, number of blood vessels formed and hemoglobin content were observed at 4.2 mg/kg of 4-AP and 20 mg/kg and 30 mg/kg of Mefloquine. Conclusions: These scientific findings indicate the use of Mefloquine and 4-Aminopyridine in pathological situations involving excessive angiogenesis. Negative regulation of cell volume, cell migration and proliferation of blood vessels may be the underlying molecular mechanisms.
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Abstract Angiogenesis plays a key role in tumor growth, invasion and metastasis of cancer diseases and therefore, the inhibition of angiogenesis can provide an important therapeutic approach in cancer diseases. This study was designed to compare the anti-angiogenic activities of the ethanolic extract of Artemisia sieberi Besser, Asteraceae, and its active substance, artemisinin in both in vitro and in vivo models. To compare cytotoxicity level of ethanolic extract of A. sieberi with artemisinin, different concentrations (1–100 µg/ml) were tested using MTT assay on human umbilical vein endothelial cells. The anti-angiogenic properties of serial concentrations of ethanolic extract of A. sieberi and artemisinin were examined on human umbilical vein endothelial cells using a three-dimensional angiogenesis assay (in vitro model) and in the chick chorioallantoic membrane assay as in vivo model. The effects of ethanolic extract of A. sieberi and artemisinin were also tested on the expression of VEGFR-1, VEGFR-2 and CD34 genes using real-time PCR. Ethanolic extract of A. sieberi and artemisinin significantly (p < 0.001) inhibited the angiogenesis in the human umbilical vein endothelial cells culture whilst the ethanolic extract of A. sieberi showed higher effect in a concentration-dependent fashion (p < 0.001). The chick chorioallantoic membrane angiogenesis was also completely inhibited by ethanolic extract of A. sieberi at concentration of 33 ng/100 µl/egg. The gene expression analysis showed that the ethanolic extract of A. sieberi and artemisinin reduced the transcription of VEGFR-1, VEGFR-2 and CD34 genes in a concentration-dependent manner. This study demonstrated that the ethanolic extract of A. sieberi is strongly able to inhibit the angiogenesis in human umbilical vein endothelial cells and chick chorioallantoic membrane models compared to the artemisinin.
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Se evaluó el efecto del incremento de la temperatura de incubación en huevos de gallina Araucana sobre la tasa de eclosión, la mortalidad embrionaria, el peso del polluelo, del saco vitelino y de los órganos internos. Los huevos (n= 426) fueron distribuidos al azar en: a) grupo Control (n=215), incubados a 37,5 °C por 21 días y b) grupo Experimental (n= 211), incubados a 39,5 °C, por 18 días, disminuyéndose a 37,5 °C los 3 días restantes. En ambos grupos la incubación se realizó con una humedad relativa constante del 55 %. La tasa de mortalidad embrionaria total fue significativamente menor (P 0,05) para el grupo Control (14,9 vs 44,1 %) y consecuentemente la tasa de eclosión fue significativamente mayor (P 0,05) en este mismo grupo (97,8 vs. 80,5 %). El peso promedio de los polluelos y del saco vitelino a las 36 h post eclosión fue significativamente diferente (P 0,05) entre ambos grupos (37,1 vs. 36,3 y 2,5 vs 6,2 g para el grupo Control y Experimental respectivamente). Por otra parte, el peso de todos los órganos internos fue menor (P 0,05) en polluelos del grupo Experimental. Se concluye que el uso de una temperatura de incubación de 39,5 C por 18 días en huevos de gallina araucana influye negativamente, aumentando la mortalidad embrionaria, disminuyendo la tasa de eclosión, el peso del polluelo y de los órganos internos, y aumentando el peso del saco vitelino.
The aim of this study was to evaluate the effect of increasing incubation temperature of Araucana hen eggs on embryo mortality, hatchability,chick, yolk sac and internal organs weight. Eggs (n= 426) were randomly distributed into: a) Control group (n= 215), which were incubated at 37.5 °C for 21 days or b) Experimental group (n= 211), which were incubated at 39.5 °C, for 18 days and then at 37.5 °C for the remaining 3 days. In both groups eggs were incubated with a relative humidity of 55 %. The embryo mortality rate was greater (P 0.05) for the experimental than the control group (44.1 vs. 14.9 % respectively). On the contrary, hatching rate was greater (P 0.05) for the control than the experimental group (97.8 vs. 80.5 % respectively). The average weight of chicks, at 36 h of age, was lower (P 0.05) for the experimental than control group (36.3 vs. 37.01 g respectively). Also, increasing incubation temperature reduced (P 0.05) internal organs weight of chicks in the Experimental compared to Control group. On the other hand, yolk sac weight was reduced (P <0.05; 2.5 vs. 6.2 g for Control and Experimental group respectively). We conclude that increasing the incubation temperature of Araucana hen eggs up to 39.5 °C for 18 days has a negative effect on embryo survival, hatchability, chick and internal organs weight and absorption of yolk.
Subject(s)
Animals , Chick Embryo , Chickens/growth & development , Temperature , Body Weight , Chickens/anatomy & histology , Eggs , Mortality , Organ SizeABSTRACT
ObjectiveTo investigate the effect of sorafenib and/or thalidomide on angiogenesis in chick chorioallantoic membrane (CAM). MethodsWhite eggs incubated for 7 days were used to establish a CAM model. The model eggs were randomly divided blank control group, sorafenib group, thalidomide group, and sorafenib/thalidomide group. After treatment, each egg was incubated for another 2 days. The CAM samples were collected and fixed to take their pictures under a microscope, and the vascular coverage of each sample was calculated. Comparison between these groups was made by analysis of variance, and comparison between each two groups was made by SNK-q test. ResultsThe thalidomide group or sorafenib group had significantly lower vascular coverage than the blank control group (30.2%±2.9% or 26.5%±2.1% vs 38.3%±2.7%, P＜0.05). The sorafenib/thalidomide group had significantly lower vascular coverage than the thalidomide group or sorafenib group (12.6%±1.5% vs 30.2%±2.9% or 26.5%±2.1%, P＜0.05). ConclusionBoth sorafenib and thalidomide have a good anti-angiogenic effect on CAM, but a combination of the two drugs shows better efficacy.