Showing papers on "Toad published in 2014"
TL;DR: The toad skins that originated from the Jiangsu province, have comparatively greater advantages over samples from other regions as far as active constituent content and potential anti-lung cancer activity is concerned, suggesting that it can be a promising chemotherapeutic agent in lung cancer therapy, in further studies.
Abstract: Background: The skin of Bufo bufo gargarizans Cantor, rich in bufadienolides, peptides, and alkaloids, has approved pharmacological activity for preliminary anti-liver and lung tumor treatment. However, few studies have systematically focused on the influence of the producing regions on the content and antitumor activity of the active constituents in toad skins. Objective: This study aims to compare toad skins obtained from six different regions in China (Jiangsu, Anhui, Henan, Hebei, Jiangxi, and Shandong province) for their bufadienolide and alkaloid content, and their cytotoxic activity on two lung carcinoma cell lines (SPC-A-1 cells and A549 cells). Materials and Methods: High performance liquid chromatography (HPLC) was used to quantificationally determine four bufadienolides, which included bufotalin, bufalin, cinobufagin, and resibufogenin in toad skins, from six different regions, respectively. In addition, an ultraviolet (UV) spectrophotometer was also employed to identify the content of the total alkaloids using 5-hydroxytryptamine (5-HT) as the reference substance. An MTT assay was performed to compare the antiproliferative effects of the toad skins' ethanolic extracts from the different regions against SPC-A-1 and A549 cells. Results: In this study, the toad skins from Jiangsu province had the highest amount of bufadienolides (472.6 μg/g crude drug) and alkaloids (1.51 mg/g crude drug). Meanwhile, according to the extract, it exhibited the strongest cytotoxic effect against the lung carcinoma cell line (SPC-A-1 cells and A549 cells) with IC 50 values of 24.82 ± 0.76 and 23.77 ± 0.63 μg crude drug/mL, respectively. Conclusion: The toad skins that originated from the Jiangsu province, have comparatively greater advantages over samples from other regions as far as active constituent content and potential anti-lung cancer activity is concerned, suggesting that it can be a promising chemotherapeutic agent in lung cancer therapy, in further studies.
15 citations
TL;DR: The findings suggest that the green toad was sensitive to the cadmium treatment, with LC50 values lower than those reported by other studies, and this species could be considered a reliable indicator species of environmental stress in aquatic ecosystem.
Abstract: The environmental impact of cadmium use and its accumulation in nature have increased to alarming levels. This study aimed to morphologically and histologically investigate the acute toxic effects of cadmium on green toad, Pseudepidalea variabilis (Pallas, 1769) larvae. Embryos were obtained from specimens collected in amplexus from nature and kept under laboratory conditions until stage 26, when they were exposed to cadmium (0, 1, 5, 10, 25, and 50 µg L -1 ) for 96 h. The LC 10, LC 50 ,and LC 90 values of cadmium were calculated to be 26.98, 35.35, and 46.31 µg L -1 , respectively. Our results showed that cadmium had a negative effect on the body size of P. variabilis larvae (over 1 µg L -1 ). Histological examination detected a fusion of gill lamellae, liver haemorrhage, oedema in the abdominal cavity, and deformations of pronephric tubules (over 10 µg L -1 ). Our findings suggest that the green toad was sensitive to the cadmium treatment, with LC 50 values lower than those reported by other studies. Thus, this species could be considered a reliable indicator species of environmental stress in aquatic ecosystem.
11 citations
TL;DR: The findings indicate that toad melanophores contain phylogenetically significant information of anatomical similarity with lower as well as higher vertebrates which can help to better understand the inter relationships between vertebrate pigment cells and their role in skin dysfunctions.
9 citations
TL;DR: The results suggest that calmodulin, colocalized with the calcium pump at the SC cytoplasmic level, would be involved in the active transport of the cation inside the secretory granules, maintaining adequate levels of intracellular Ca(2+) .
Abstract: The presence of a calcium pump, calbindin D-28KD, and calmodulin in the secretory cells (SC) of the oviductal pars convoluta (PC) of Rhinella arenarum was established for the first time in amphibians using immunohistochemical techniques Marked variations were observed in the localization and degree of expression of these proteins according to the duct segment and the period of the sexual cycle analyzed During the preovulatory and ovulatory periods the calcium pump colocalized with calbindin D-28KD can be seen mainly in the apical border of the SC, which are located in the first zones of PC and synthesize and secrete the components of the inner jelly coat layers These envelopes, which surround the oocytes, contain the molecules indispensable for fertilization, probably inducing the sperm acrosome reaction (AR) Our results suggest that calmodulin, colocalized with the calcium pump at the SC cytoplasmic level, would be involved in the active transport of the cation inside the secretory granules, maintaining adequate levels of intracellular Ca2+ During the postreproductive period, a calcium pump colocalized with calbindin D-28KD appears for the first time in the cycle in the basal zones of the SC This system may be related to the replenishing of intracellular Ca2+ stores In contrast, in R arenarum the Ca2+ present in the jelly coats that surround the oocytes participates in the AR during fertilization, suggesting that this secretion system of the cation provided by the oviductal mucosa is functionally more active during the reproductive period of this species J Exp Zool 321A: 432–441, 2014 © 2014 Wiley Periodicals, Inc
4 citations
TL;DR: The recorded remodeling of the small intestine represents a model for clarifying the mechanism whereby cell death and proliferation are controlled and is investigated using twofold approaches, namely, ultrastructural and molecular.
Abstract: The ontogenetic development of the small intestine of the toad Bufo regularis was investigated using twofold approaches, namely, ultrastructural and molecular. The former has been done using transmission electron microscope and utilizing the developmental stages 42, 50, 55, 60, 63, and 66. The most prominent ultrastructural changes were recorded at stage 60 and were more evident at stage 63. These included the appearance of apoptotic bodies/nuclei within the larval epithelium, the presence of macrophages, swollen mitochondria, distorted rough endoplasmic reticulum, chromatin condensation, and irregular nuclear envelop, and the presence of large vacuoles and lysosomes. The molecular investigation involved examining DNA content and fragmentation. The results showed that the DNA content decreased significantly during the metamorphic stages 60 and 63 compared with both larval (50 and 55) and postmetamorphic (66) stages. The metamorphic stages (60 and 63) displayed extensive DNA laddering compared with stages 50, 55, and 66. The percentage of DNA damage was 0.00%, 12.91%, 57.26%, 45.48%, and 4.43% for the developmental stages 50, 55, 60, 63, and 66, respectively. In conclusion, the recorded remodeling of the small intestine represents a model for clarifying the mechanism whereby cell death and proliferation are controlled.
4 citations
TL;DR: The influence of temperature on mortality and development rate of common toad Bufo bufo was investigated and it was shown that the rate of embryogenesis at +19°C increased almost 1.4 times as compared with the development of a temperature of +15°C.
Abstract: The influence of temperature on mortality and development rate of common toad Bufo bufo was investigated in the laboratory conditions. It was shown that the rate of embryogenesis at +19°C increased almost 1.4 times as compared with the development of a temperature of +15°C. The temperature affects the resulting embryo mortality at the time of hatching primarily in clutches with low survival.
3 citations
TL;DR: Investigation of the distribution and density of six types of gastrointestinal (GI) hormone immunoreactive (IR) cells in the skin and parotoids of Bufo gargarizans found 5-HT-IR cells were distributed in all layers of epidermis and glands, with higher density in the glands.
Abstract: The skin and skin secretion of Chinese toad Bufo gargarizans have long been used in traditional Chinese medicine. However, the exact types and location of bioactive substances in Bufo gargarizans skin still have not been fully elucidated. The aim of the study was to investigate the distribution and density of six types of gastrointestinal (GI) hormone immunoreactive (IR) cells in the skin and parotoids of Bufo gargarizans . Immunohistochemistry was used for qualitative and semiquantitative analysis of GI hormone presence in the dorsal and ventral skin, and parotoids of eight adult Chinese toads. Six types of IR cells were found: serotonin (5-HT), glucagon (GLU), gastrin (GAS), somatostatin (SS), pancreatic polypeptide (PP) and neuropeptide Y(NPY) IR cells. They were mainly present in the epidermis and skin glands. 5-HT-IR cells were distributed in all layers of epidermis and glands, with higher density in the glands. Glucagon was prominently expressed in the epidermis and the bottle-shaped glands of parotoids; however, it was not present in the granular glands of skin and parotoids. The distributions of GAS and SS-IR cells were similar since they were present mainly in mucous, granular and bottle-shaped glands, while these cell types were absent in the differentiated glands of parotoids. PP-IR cells were predominant in the granular glands and the bottle-shaped glands. The expression of NPY was high in epidermal stratum granulosum and mucous glands of the dorsal skin, the bottle-shaped glands and differentiated glands of parotoids, while NPY-IR was rarely seen in the granular glands of ventral skin, and not present in the granular glands of dorsal skin and parotoids. The expression of several types of GI hormones in the skin and parotoids of Bufo gargarizans varies depending on tissue and type of glands.
3 citations
Patent•
29 Jan 2014
TL;DR: In this article, a cloning method of toad skin calmodulin binding protein EDF-1 Endothelial Differentiation-related Factor-1) coding gene for inhibiting endothelial cell differentiation, belonging to the technical field of biological medicines.
Abstract: The invention discloses a cloning method of toad skin calmodulin binding protein EDF-1 Endothelial Differentiation-related Factor-1) coding gene for inhibiting endothelial cell differentiation, belonging to the technical field of biological medicines. PCR (Polymerase Chain Reaction) is performed by taking first chain cDNA (complementary Deoxyribonucleic acid) of skin of the toad; the obtained PCR product is converted into E.coli competent cell; the toad skin calmodulin binding protein EDF-1 coding gene capable of being used for inhibiting the endothelial cell differentiation is obtained through screening of positive colonies, plasmid recovery and enzyme digestion. The cloning method has a simple and convenient process and is sufficient in raw materials, and the obtained coding gene has stable performance.
2 citations
Patent•
26 Feb 2014
TL;DR: In this paper, a method for preparing a medical toad cortex preparation is described, which includes spraying edible vinegar on the surface of a toad body, leaving the toad in a dry activity room after 3-5 days, and immediately taking out the cortex which is naturally sloughed off by the toads; removing impurities of the collected toads cortex, drying the cortex in a drying tank, irradiating the cortex with ultraviolet ray for sterilization until the moisture content is less than or equal to 5 percent.
Abstract: The invention discloses a method for preparing a medical toad cortex preparation. The method comprises the following steps: spraying edible vinegar on the surface of a toad body, leaving the toad in a dry activity room after 3-5 days, and immediately taking out the toad cortex which is naturally sloughed off by the toad; removing impurities of the collected toad cortex, drying the toad cortex in a drying tank, irradiating the toad cortex with ultraviolet ray for sterilization until the moisture content is less than or equal to 5 percent; crushing the dried and sterilized toad cortex, and screening through a screen with the granularity of 50-100 meshes to obtain toad cortex coarse powder; crushing once again to obtain toad cortex fine powder with the granularity of 800-1,000 meshes; mixing and stirring the toad cortex fine powder with the granularity of 800-1,000 meshes and tolypocladium sinensis mycelia with the granularity of 600-800 meshes, and continuously drying the mixture in the drying tank until the moisture content is less than 2 percent. According to the method, the toad body naturally sloughs off the toad cortex, and the content of effective components is high; chemicals are not used in a toad cortex sloughing process, so that the pollution is small, and a higher effect can be maintained; the toad cortex and the tolypocladium sinensis mycelia are combined with each other, so that the absorption is accelerated, and the anti-cancer effect can be strengthened.
1 citations
TL;DR: Tissue distribution analysis showed thatSCP-2 expressed in all organs tested, though in the liver and spleen it manifested lower expression than in other organs, which might indicate SCP-2 being one of the active ingredients in toad skin.
Abstract: In this study, to clarify the bioactive polypeptides included in the skins and secretions of Bufo, we screened the Japanese toad (Bufo japonicus formosus) skin cDNA library by colony polymerase chain reaction (PCR), and obtained a transcript of 1 075 bp consisting of 1 37 bp 5' untranslated region (UTR), 515 bp 3' UTR and a 423 bp open reading frame (ORF) encoding a polypeptide of 140 amino acid residues (GenBank accession number: KF359945). Homolog analysis showed a 70%-96% homology with sterol carrier protein-2 (SCP-2) present in other animals, which is implicated in lipid metabolism of other organisms. The gene SCP-2 of Chinese toad (B. gargarizans) was cloned from a first strand cDNA of Bufo skin (GenBank accession number: KF381341) via PCR, whose encoding polypeptide has only one amino acid difference from that of Japanese toad. Tissue distribution analysis showed that SCP-2 expressed in all organs tested, though in the liver and spleen it manifested lower expression than in other organs. These findings might indicate SCP-2 being one of the active ingredients in toad skin. These findings may in turn have implications for further drug development from traditional Chinese medicine sources.
1 citations
TL;DR: It is found that vitellogenic cycle is continuous in toad, but discontinuous in frog, and that higher sexual activities occurred in spring for toad but in moist summer for frog, while eggs of the frog are controlled by both temperature and humidity in summer season.
Abstract: I examined the reproductive strategies of leopard toad and mascarene frog by studying their annual vitellogenic cycle, monthly changes of masses of ovary, liver and fat bodies as well as egg size and number in two study areas, Abo Roash and El Mansuriya, and in the years 2001, 2005, and 2008–2009, particularly during the final two years of that period. Based on the presence of the mature ova, I found that vitellogenic cycle is continuous in toad, but discontinuous in frog. Further, leopard body reserves allocated more energy to vitellogenesis than did mascarene frog. Hence, fecundity in toad was higher than that in frog, as associated with higher egg number and size. During oviposition, female mascarene retained a small portion of a clutch, whereas toad shed all egg mass at once. Over the study period, both body and reproductive conditions reacted positively in toad, but negatively in frog. Warm temperature and long photoperiod elucidated ovarian development under high relative humidity in frog. In contrast, in toad, low relative humidity may be an environmental cue for the increase in ovarian mass. Thus, higher sexual activities occurred in spring for toad (dry environment), but in moist summer for frog. Ovarian mass and egg number were temperature-dependent in frog, but independent in toad. Relative humidity correlated significantly and negatively to egg size in both populations. It also related inversely to egg number in toad, but not in frog. Hence, eggs of the frog are controlled by both temperature and humidity in summer season. Rainfall had no effect on sexual parameters in both species.
TL;DR: It is reported that the parotoid gland secretion increased the force of contraction, heart beat and cardiac output in perfused frog’s heart, whereas, there was no change on hypodynamic heart, indicating that there may be existence of two components, one with β–receptor stimulating activity and other acting directly on the heart (independent of β1-adrenoreceptors).
Abstract: The present investigation has been undertaken to study the cardiotonic activity of parotoid gland secretion of common Indian Toad Bufo melanostictus on isolated perfused frog heart (Rana tigrina). Cardiotonic activity of parotoid gland secretion of B. melanostictus in normal as well as when induced with paraoxon (an organophosphate) was studied. For the evaluation of cardiotonic activity, Syme’s technique is being employed, digoxin and propranolol were used as standard drug and β-blocker respectively to characterize the effects on the receptors. The isolated perfused frog heart and hypodynamic frog heart showed dose dependent positive ionotrophic effects. The parotoid gland secretion exihibited cardiac stimulant activities. The propranolol was not able to block the effects of paraoxon induction on toad secretion. Thus, the present investigation reports that the parotoid gland secretion increased the force of contraction, heart beat and cardiac output in perfused frog’s heart, whereas, there was no change on hypodynamic heart, indicating that there may be existence of two components, one with β–receptor stimulating activity and other acting directly on the heart (independent of β1-adrenoreceptors).
TL;DR: The blowfly Lucilia bufonivora shows high host specificity for toads despite the host’s toxic skin secretion, which consists mainly of bufadienolides, and alternative mechanisms of resistance such as efficient excretion of the compounds may enable the flies to use this poisonous food source.
Abstract: The blowfly Lucilia bufonivora shows high host specificity for toads despite the host’s toxic skin secretion, which consists mainly of bufadienolides. These toxins are effective blockers of the Na+, K+-ATPase, an enzyme that is essential for many physiological processes in animals. Whereas common toad (Bufo bufo) toxins were identified in the larvae of the fly, few toxins were found in the pupae and empty puparia as trace amounts, while adult flies were entirely free of these toxic compounds. Similar results were obtained when larvae of generalist necrophagous blowflies (L. sericata, Calliphora vicina) fed on toad carcasses. Analysis of the Na+, K+-ATPase gene revealed no amino acid substitution at positions known to mediate resistance to bufadienolides in other systems. Alternative mechanisms of resistance such as efficient excretion of the compounds may enable the flies to use this poisonous food source.
01 Jan 2014
TL;DR: Meshed ventricular tissue of toad tadpoles was employed as explant and inoculated in culture medium supplemented with vitamin A, showed good regeneration ability with rhythmic beating and Vitamin A was found good model to accelerate the percentage of cardiac tissue regeneration.
Abstract: Present findings provide valuable basic information on cardiac tissue regeneration in culture medium. Meshed ventricular tissue of toad tadpoles was employed as explant and inoculated in culture medium supplemented with vitamin A , showed good regeneration ability with rhythmic beating. Vitamin A was found good model to accelerate the percentage of cardiac tissue regeneration. The culture system appears to be a suitable for investigating the changes occurred during regeneration and differentiation of cardiac muscles.