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In this study, we investigated the potential in vitro anti-HSV-1 activities of the Cassiopea andromeda jellyfish tentacle extract (TE) and its fractions, as well as computational work on the thymidine kinase (TK) inhibitory activity of the identified secondary metabolites. The LD50, secondary metabolite identification, preparative and analytical chromatography, and in silico TK assessment were performed using the Spearman-Karber, GC-MS, silica gel column chromatography, RP-HPLC, LC-MS, and docking methods, respectively. The antiviral activity of TE and the two purified compounds Ca2 and Ca7 against HSV-1 in Vero cells was evaluated by MTT and RT-PCR assays. The LD50 (IV, mouse) values of TE, Ca2, and Ca7 were 104.0 ± 4, 5120 ± 14, and 197.0 ± 7 (µg/kg), respectively. They exhibited extremely effective antiviral activity against HSV-1. The CC50 and MNTD of TE, Ca2, and Ca7 were (125, 62.5), (25, 12.5), and (50, 3.125) µg/ml, respectively. GC-MS analysis of the tentacle extract revealed seven structurally distinct chemical compositions. Four of the seven compounds had a steroid structure. According to the docking results, all compounds showed binding affinity to the active sites of both thymidine kinase chains. Among them, the steroid compound Pregn-5-ene-3,11-dione, 17,20:20,21 bis [methylenebis(oxy)]-, cyclic 3-(1,2-ethane diyl acetal) (Ca2) exhibited the highest affinity for both enzyme chains, surpassing that of standard acyclovir. In silico data confirmed the experimental results. We conclude that the oxosteroid Ca2 may act as a potent agent against HSV-1.
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Venenos de Cnidários , Herpesvirus Humano 1 , Chlorocebus aethiops , Animais , Camundongos , Antivirais/farmacologia , Antivirais/química , Células Vero , Timidina Quinase/genética , Timidina Quinase/química , Venenos de Cnidários/farmacologia , Esteroides/farmacologiaRESUMO
This study was aimed to survey toxicity of waterpipe wastes leachates on Peronia peronii in aquatic and sediment environments as two exposure media. For this, leachates of four tobacco types including burnt traditional tobacco (BTT), fresh traditional tobacco (FTT), burnt fruit-flavored tobacco (BFT) and fresh fruit-flavored tobacco (FFT)) were prepared and used to assess their toxic effects on P. peronei in two aquatic and sediment media. The in-vivo toxic effects of five different concentrations of waterpipe tobacco waste leachates on P. peronii were evaluated. The LC50 values of BTTs leachates to P. peronii were 17.50, 16.05, 11.31 and 9.38 g/L at exposure times of 24, 48, 72 and 96 h, respectively in aquatic media. These values for BFTs leachates were 14.86, 12.38, 9.53 and 7.46 g/L at exposure times of 24, 48, 72 and 96 h, respectively. In the case of sediment media, the LC50 values of BTTs leachates were 15.33, 13.70, 9.09 and 6.70 g/L at exposure times of 24, 48, 72 and 96 h, respectively while these values for BFTs leachates were 12.00, 10.32, 8.20 and 5.65 g/L. Fruit-flavored tobacco leachates had significantly higher toxicity than traditional tobacco leachates for P. peronii. The findings also showed significant differences between the LC50 values of different leachates in different media of water and sediment. The results demonstrated that even small amount of tobacco waste (~ 5 to 6 g/L) can lead to P. peronii mortality and may also pose a hazard to other aquatic and benthic organisms. The results obtained from the present study can be used as a baseline data to assess local effects causing from unsafe disposal of post-consumption tobacco waste in beach areas. In addition, these findings can lead to encouraging decision-makers to focus more on the types of tobacco waste in the municipal solid waste management system and to implement a source separation process for these wastes.
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Sedimentos Geológicos , Tabaco para Cachimbos de Água , Poluentes Químicos da Água , Poluentes Químicos da Água/toxicidade , Poluentes Químicos da Água/análise , Sedimentos Geológicos/química , Tabaco para Cachimbos de Água/toxicidade , Animais , Dose Letal Mediana , Monitoramento Ambiental/métodos , Alismatales/químicaRESUMO
INTRODUCTION: Melanoma is known as an aggressive and highly lethal cancer. The poor prognosis and resistance to treatment are characteristics of melanoma. In melanoma cells, apoptosis signaling which relies heavily on the acute activity of mitochondria and reactive oxygen species (ROS) formation is suppressed. Studies have shown that compounds isolated from marine herbs and animals, have been shown to have cytotoxic consequences on cancerous cells in prior research. This study was designed to evaluate the apoptotic effect of methanolic extract of Persian Gulf shell-less marine mollusc (Peronia peronii) on skin mitochondria isolated from animal model of melanoma. PURPOSE: Melanoma mitochondria obtained from skin of melanoma animal model are studied in this research to see whether extracts from Persian Gulf shell-less marine mollusc (Peronia peronii), has a cytotoxic impact on them. MATERIAL AND METHOD: In this study, the mitochondria were isolated from melanoma cells via differential centrifugation were treated with various concentrations (650, 1300 and 2600 µg/ml) of methanolic extract of Peronia peronii. Then MTT(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) viability assay, Reactive oxygen species (ROS) determination, Mitochondrial Membrane Potential (MMP) decline assay, mitochondrial swelling and cytochrome c release determination were performed. Flow cytometry assay of % apoptotic vs necrotic phenotypes was also performed on extract treated melanoma cells. RESULTS: The results of MTT assay showed that different concentrations of Peronia peronii extract significantly (P < 0.05) decreased the SDH activity in cancerous skin mitochondria with the IC50(1300 µg/ml). The ROS results also showed that all concentrations of Peronia peronii extracts significantly increased ROS production, MMP decline and the release of cytochrome c in cancer groups mitochondria. The swelling of mitochondria was significantly increased compared to the control group. In addition, the results of apoptosis assay showed that addition of root extract of Peronia peronii on melanoma cells increased apoptosis, while it had no effect on control non tumour cells. DISCUSSION AND CONCLUSION: Based on these results, the presence of potentially bioactive compounds in Peronia peronii make this Persian Gulf coastal herb a strong candidate for further molecular studies and clinical research in the field of melanoma cancer therapy.
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Antineoplásicos , Melanoma , Animais , Espécies Reativas de Oxigênio , Oceano Índico , Citocromos c , Melanoma/tratamento farmacológico , Melanoma/patologia , Mitocôndrias , Antineoplásicos/farmacologia , MoluscosRESUMO
INTRODUCTION: As a major public health issue, skin cancer is a leading reason of death and has resulted in significant financial and human losses globally. Numerous environmental and internal variables may both drive and exacerbate the pathophysiology of skin cancer. Marine herbs and animals, including marine sponges, cucumbers, and squirts, have been shown to have cytotoxic consequences on cancerous cells in prior research. PURPOSE: melanoma mitochondria obtained from the skin of melanoma animal models are studied in this research to see whether extracts from Cistanche tubulosa, a plant endemic to the northern coasts of the Persian Gulf, have a cytotoxic impact on them. MATERIAL AND METHOD: In this study, the mitochondria were isolated from melanoma cells via differential centrifugation and treated with various concentrations (1250, 2500 and 5000 µg/ml) of methanolic extract of C. tubulosa. Then MTT, ROS, MMP decline, mitochondrial swelling, cytochrome c release and flow cytometry assays were performed on them. RESULTS: The results of the MTT assay showed that the IC50 of C. tubulosa extract is 2500 µg/ml and C. tubulosa extract induced a selectively significant (P < 0.05) concentration-dependent decrease in the SDH activity in cancerous skin mitochondria. The ROS results also showed that all concentrations of C. tubulosa extracts significantly increased ROS production, MMP decline and the release of cytochrome c in cancer group mitochondria. The swelling of mitochondria isolated from the cancer group was significantly increased compared to the control group. In addition, the results of the apoptosis assay showed that the addition of root extract of C. tubulosa on melanoma cells increased apoptosis, while it had no effect on control non-tumour cells. DISCUSSION AND CONCLUSION: Based on these results, the presence of potentially bioactive compounds in C. tubulosa makes this Persian Gulf coastal herb a strong candidate for further molecular studies and clinical research in the field of melanoma cancer therapy.
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Cistanche , Melanoma , Neoplasias Cutâneas , Animais , Citocromos c , Modelos Animais de Doenças , Humanos , Melanoma/tratamento farmacológico , Mitocôndrias , Extratos Vegetais/farmacologia , Extratos Vegetais/uso terapêutico , Espécies Reativas de Oxigênio , Neoplasias Cutâneas/tratamento farmacológicoRESUMO
Stonustoxin (SNTX) is a lethal protein found in stonefish venom, responsible for many of the symptoms associated with stonefish envenomation. To counter stonefish venom challenges, antivenom is a well-established and effective solution. In this study, we aimed to produce the recombinant alpha subunit protein of Stonustoxin from Synanceia horrida and prepare antibodies against it The SNTXα gene sequence was optimized for E. coli BL21 (DE3) expression and cloned into the pET17b vector. Following purification, the recombinant protein was subcutaneously injected into rabbits, and antibodies were extracted from rabbit´s serum using a G protein column As a result of codon optimization, the codon adaptation index for the SNTXα cassette increased to 0.94. SDS-PAGE analysis validated the expression of SNTXα, with a band observed at 73.5 kDa with a yield of 60 mg/l. ELISA results demonstrated rabbits antibody titers were detectable up to a 1:256,000 dilution. The isolated antibody from rabbit´s serum exhibited a concentration of 1.5 mg/ml, and its sensitivity allowed the detection of a minimum protein concentration of 9.7 ng. In the neutralization assay the purified antibody against SNTXα protected mice challenged with 2 LD50. In conclusion, our study successfully expressed the alpha subunit of Stonustoxin in a prokaryotic host, enabling the production of antibodies for potential use in developing stonefish antivenom.
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Proteínas Recombinantes , Animais , Coelhos , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/genética , Proteínas Recombinantes/biossíntese , Camundongos , Antivenenos/imunologia , Antivenenos/biossíntese , Antivenenos/genética , Venenos de Peixe/imunologia , Venenos de Peixe/genética , Venenos de Peixe/química , Escherichia coli/genética , Escherichia coli/metabolismo , Expressão Gênica , Soros Imunes/imunologiaRESUMO
Introduction: The Persian Gulf is home to a diverse range of marine life, including various species of fish, crustaceans, mollusks, and echinoderms. This study investigates the potential therapeutic properties of venoms from echinoderms in the Persian Gulf, specifically their ability to inhibit cholinesterases (Acetylcholinesterase and butyrylcholinesterase) and act as antioxidants. Methods: Four venoms from two echinoderm species, including the spine, gonad, and coelomic fluids of sea urchins, as well as brittle star venoms, were analyzed using various methods, including LD50 determination, protein analysis, antioxidant assays, GC-MS for secondary metabolite identification, and molecular docking simulations. Results and discussion: The study's results revealed the LD50 of the samples as follows: 2.231 ± 0.09, 1.03 ± 0.05, 1.12 ± 0.13, and 6.04 ± 0.13 mg/mL, respectively. Additionally, the protein levels were 44.037 ± 0.002, 74.223 ± 0.025, 469.97 ± 0.02, and 104.407 ± 0.025 µg/mL, respectively. SDS-PAGE and total protein studies indicated that at least part of the venom was proteinaceous. Furthermore, the study found that the brittle star samples exhibited significantly higher antioxidant activity compared to other samples, including the standard ascorbic acid, at all tested concentrations. GC-MS analysis identified 12, 23, 21, and 25 compounds in the samples, respectively. These compounds had distinct chemical and bioactive structures, including alkaloids, terpenes, and steroids. Conclusion: These venoms displayed strong cholinesterase inhibitory and antioxidant activities, likely attributed to their protein content and the presence of alkaloids, terpenes, and steroids. Notably, the alkaloid compound C 7 was identified as a promising candidate for further research in Alzheimer's disease therapy. In conclusion, echinoderms in the Persian Gulf may hold significant potential for discovering novel therapeutic agents.
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Cigarette butts (CBs) are one of the most common, long-lasting, and toxic forms of marine and coastal area debris. Although the significance of CBs and the toxic contents of this waste items are well recognized, but there is still a lack of information about the effects of this waste on the aquatic organisms. Therefore in this study, the in-vivo toxic effects of various CBs leachates (smoked cigarette butts with tobacco [SCBs], smoked CBs without tobacco [SFs], and unsmoked filters [USFs]) on cellular and chemical hematologic markers in fish (Periophthalmus waltoni) were evaluated. In three acute, sub-acute, and sub-chronic exposure measurements, P. waltoni exposed to different CBs leachates showed a significant increase in white blood cells, creatine kinase, lactate dehydrogenase, alkaline phosphatase, alanine transaminase, and aspartate transaminase, as well as a decrease in hemoglobin (Hb) levels. The mean ± SD values of Hb in P. waltoni species exposed to different CBs leachates (control, SCBs, SFs and USFs) in acute (1 day) phase were 7.15 ± 0.34, 6.02 ± 0.29, 6.25 ± 0.25 and 6.89 ± 0.1 g/dl respectively. These values in subacute (28 days) phase were 6.70 ± 0.15, 5.19 ± 0.24, 5.67 ± 0.30 and 6.10 ± 0.24 g/dl and in sub chronic exposure (42 days) phase were 7.20 ± 0.40, 5.16 ± 0.30, 5.88 ± 0.34 and 6.60 ± 0.33 g/dl respectively. Our results showed that CBs leachates act as a stressor, leading to changes in some hematologic markers in P. waltoni species. Because of the continued deposition of CBs waste into global aquatic environments, policies to mitigate this waste in coastal areas are needed to prevent potentially negative effects on fish and other aquatic organisms.
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Perciformes , Produtos do Tabaco , Animais , Oceano Índico , Nicotiana , Peixes , Fumaça , Organismos AquáticosRESUMO
Cigarette butts (CBs) are one of the most commonly found types of litter contaminating the aquatic environment. However, the environmental risks posed by CBs need further investigation. In this study, the in-vivo toxic effects of various concentrations of CB leachates on juvenile (5.45 ± 1.36 gr and 7.08 ± 1.12 cm) fish (Periophthalmus waltoni) were evaluated. The LC50 values of CB leachate from smoked cigarette butts with tobacco (SCB) were 3.75, 3.0, 1.94, and 1.37 CBs/L in 24, 48, 72, and 96 h exposure times, respectively. The LC50 values for leachate of smoked CBs without tobacco (SF) were 7.58, 6.22, 4.73, and 2.9 CBs/L at 24, 48, 72, and 96 h exposure times, respectively. In the case of leachate from unsmoked filters (USF), LC50 values were 14.68, 12.44, 10.19, and 7.46 CBs/L in 24, 48, 72, and 96 h exposure time, respectively. The mean concentrations of heavy metals and polycyclic aromatic hydrocarbons in SCBs leachates were higher than in SF and USF leachates. Our findings report that even low concentrations of CBs leachates can led to lethality of P. waltoni and may pose a threat to their population density.
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OBJECTIVE: Compounds isolated from marine animals have different pharmacological effects. In this study, we investigated the effects of sea nettle (Chrysaora quinquecirrha) crude venom on human colon cancer mitochondria. METHODS: First, mitochondria were isolated from healthy colon tissue and cancerous colon tissue, and then mitochondrial function (SDH activity), reactive oxygen species (ROS) level, mitochondrial membrane potential (MMP) collapse, mitochondrial swelling, and cytochrome c release were measured. RESULTS: The results showed that crude venom of Chrysaora quinquecirrha (180, 360 and 720 µg/ml) can significantly impair mitochondrial function (**P<0.01 and ***P<0.001) and consequently increase the level of ROS (*P<0.05 and ****P<0.0001), collapse in MMP (*P<0.05 and ****P<0.0001), mitochondrial swelling (**** P<0.0001) and release of cytochrome c (* P<0.05 and *** P<0.001) only in mitochondria isolated from human colon cancer tissue. CONCLUSION: The results concluded that crude venom of Chrysaora quinquecirrha (180, 360 and 720 µg/ml) has no side effects on normal mitochondria and only selectively affects cancerous mitochondria. It seems that after further research, Chrysaora quinquecirrha can be considered as a drug candidate for the treatment of patients with colon cancer.
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Antineoplásicos/farmacologia , Neoplasias Colorretais/tratamento farmacológico , Mitocôndrias/efeitos dos fármacos , Urtiga-do-Mar da Costa Leste/química , Peçonhas/farmacologia , Animais , Colo/metabolismo , Citocromos c/metabolismo , Humanos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Reto/metabolismoRESUMO
BACKGROUND: Research on animal toxins have shown toxicity potential on cancerous cell and tissues in the cultures. Conotoxins obtained from marine cone snails show the highest toxicity potential, so that several human deaths have been attributed to this species of snail. These toxins have proven to be valuable agents to inhibit enzymes, channels and proteins, in the nervous systems of humans. METHODS: We have studied the effects of Conus textile crude venom on U87MG human glioma cells and their mitochondria as main inducers of apoptosis and human embryonic kidney 293 cells (HEK293) as non-cancerous normal control cells. Cellular toxicity assessments including MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay and measurement of caspase-3 activation as well as mitochondrial toxicity assays including measurement of the activity of succinate dehydrogenase (SDH) enzyme, mitochondrial swelling, reactive oxygen species (ROS) production, collapse of mitochondrial membrane potential (MMP) and cytochrome c release were performed in U87MG human glioma cells and HEK293 cells (as non-cancerous normal cells). RESULTS: The results illustrated the significant cytotoxic effect of Conus textile crude venom on U87MG human glioma cells, that inhibits 50% (IC50=10µg/mL) of the cell growth after 12 h of exposure. Viability measurement showed which the Conus textile crude venom is selectively cytotoxic to U87MG human glioma cells, and induced activation of caspase-3 and induction of cell apoptosis via through mitochondrial signaling. Conus textile crude venom also selectively increased mitochondria swelling, ROS formation, cytochrome c release and MMP decrease in cancerous mitochondria but not normal mitochondria. Conclusion; Based on the obtained results from this investigation, it is concluded that the Conus textile crude venom contains promising natural compounds to fight U87MG human glioma cells through activation of apoptosis intrinsic pathways.
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Antineoplásicos/farmacologia , Apoptose , Glioblastoma/tratamento farmacológico , Mitocôndrias/efeitos dos fármacos , Venenos de Moluscos/farmacologia , Animais , Proliferação de Células , Glioblastoma/metabolismo , Glioblastoma/patologia , Humanos , Mitocôndrias/metabolismo , Mitocôndrias/patologia , Espécies Reativas de Oxigênio/metabolismo , Células Tumorais CultivadasRESUMO
BACKGROUND: Marine animals have been considered by many researchers due to their various pharmacological effects. One group of marine animals that have been studied is cone snails. The conotoxin obtained from these marine animals has various therapeutic effects. METHODS: This study was designed to investigate the apoptotic effects of crude venom of Conus textile and its fractions (A and B) on chronic lymphocytic leukemia (CLL) cells. Accordingly, parameters such as cell viability, reactive oxygen species (ROS) level, collapse in mitochondrial membrane potential (MMP), lysosomal membrane damage and caspase-3 activation were evaluated. RESULTS: The results showed that the crude venom (50, 100 and 200 µg/ml) from Conus textile and its fraction B (50, 100 and 200 µg/ml) significantly reduced viability in CLL B-lymphocyte. In addition, exposure of CLL B-lymphocyte to fraction B (50, 100 and 200 µg/ml) was associated with an increase in the level of ROS, the collapse of the MMP, damage to the lysosomal membrane, and activation of caspase-3. CONCLUSION: According to results, it was concluded that fraction B from crude venom of Conus textile causes selective toxicity on CLL B-lymphocyte with almost no effect on a normal lymphocyte. Furthermore, this venom fraction could be a promising candidate for induction of apoptosis in patients with CLL through the mitochondrial pathway.
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Caspase 3/metabolismo , Caramujo Conus/química , Leucemia Linfocítica Crônica de Células B/patologia , Linfócitos/patologia , Mitocôndrias/efeitos dos fármacos , Peçonhas/farmacologia , Animais , Apoptose , Sobrevivência Celular , Células Cultivadas , Humanos , Leucemia Linfocítica Crônica de Células B/tratamento farmacológico , Leucemia Linfocítica Crônica de Células B/metabolismo , Linfócitos/efeitos dos fármacos , Potencial da Membrana Mitocondrial , Espécies Reativas de Oxigênio/metabolismoRESUMO
Cholinesterase inhibitors find application in the combat and care of several diseases, especially AD. Jellyfish venoms are the most promising sources of potent cholinesterase inhibitors. Therefore, it is of interest to study cholinesterases inhibiting compounds from the Cassiopea andromeda venom. We report bioactive compounds using the GC-MC method followed by molecular modeling and docking data analysis. The GC-MS analysis of the crude venom led to the identification of seven bioactive compounds (C1-C7), comprising the steroidal alkaloids, phenolic and carotenoid derivatives. The venom exhibited inhibitory activities against the cholinesterase enzymes. The compound C2, a Dioxolane steroid, displayed the strongest inhibition on both AChE and BChE activities for further consideration.
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Despite recent improvements in treatment, ovarian cancer is still the leading cause of death from gynaecological malignancies. Today, marine mollusks are considered as natural source of new biologically and pharmacologically active compounds by scientists and the pharmaceutical industries. The aim of this study is to investigate the selective apoptotic effects of Turbo coronatus crude extract fractions on human epithelial ovarian cancer (EOC) cells and mitochondria. The cells and mitochondria were isolated from cancerous and non-cancerous ovarian tissues and exposed to IC50 concentration of F1 fraction for evaluation of mitochondrial and cellular parameters. Our results showed that F1 fraction of T. coronatus crude extract significantly induced toxic effects only in the cancerous ovarian mitochondria, including increased reactive oxygen species (ROS) formation, mitochondrial membrane depolarization, mitochondrial swelling, and cytochrome c release.Flow-cytometry analysis demonstrated that F1 fraction of T. coronatus progressively induced apoptosis and necrosis only on EOC but not non-cancerous cells. We eventuallyconcluded that F1 fraction of T. coronatus crude extract selectively induces apoptosis in EOC through a ROS- mediated pathway.
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Objective: A variety of approaches such as surgery, chemotherapy, radiotherapy, hormonal therapy and immunotherapy are used to treat melanomas, but unfortunately in most case, the response is very weak and often side effects are serious. This study concerns selective toxicity of an extract of Turbo coronatus on cells and mitochondria from a syngeneic mouse model of melanoma. Methods: Cells and mitochondria isolated from extra tumoral and melanoma tissues were exposed toa T. coronatus crude extract and fractions obtained by gel-filtration chromatography and assayed for mitochondrial and cellular parameters. Result: Crude extract (375, 750 and 1,500 µg/ml) and fraction 1; F1; (275, 550 and 1100 µg/ml) of T. coronatus extract induced a significant (p<0.05) increase of the reactive oxygen species (ROS) level, swelling of mitochondria, collapse of mitochondrial membrane potential (MMP), release of cytochrome c and caspase-3 activation only in the mitochondria and cells obtained from melanoma but not extra tumoral tissues. In addition, the F1 fraction decreased the percentage of viable cells and induced apoptosis in melanoma cells. Conclusion: For the first time we could demonstrate that the F1 fraction of a T. coronatus extract, selectively induces ROS mediated cytotoxicity by directly targeting mitochondria in melanoma tissues and it may be a suitable candidate for novel drug treatment of malignant melanomas.
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Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Melanoma/tratamento farmacológico , Moluscos/química , Espécies Reativas de Oxigênio/metabolismo , Animais , Caspase 3/metabolismo , Citocromos c , Feminino , Oceano Índico , Melanoma/metabolismo , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos C57BL , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Células Tumorais CultivadasRESUMO
For the first time, we previously recorded an enormous population of the Cassiopea andromeda jellyfish that had increased dramatically from Bushehr coasts of Iran. The sub-acute toxicity of the jellyfish venom in rat organs was correspondingly carried out. The data presented in this paper relate to the in vivo and in vitro hematological effects of this venomous species of jellyfish venom.
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Objective: This study was conducted to investigate whether fractions of jellyfish Cassiope andromeda venom, could selectively induce toxicity on mitochondria isolated from cancer tissue of patients with breast adenocarcinomas. Methods: Firstly, we extracted two fractions, (f1 and f2) from crude jellyfish venom by gel filtration on Sephadex G-200.Then different dilutions of these extracted fractions were applied to mitochondria isolated from human breast tumoral- and extra-tumoral tissues. Parameters of mitochondrial toxicity including generation of reactive oxygen species (ROS), mitochondrial membrane potential (MMP) collapse, swelling, cytochrome c release, activation of caspase3 and apoptosis were then assayed. Result: Our results demonstrate that fraction 2 of Cassiopea andromeda crude venom significantly (P<0.05) decreased mitochondrial succinate dehydrogenase activity, increased mitochondrial ROS production, induced mitochondrial swelling, MMP collapse and cytochrome c release, activated caspase3 and induced apoptosis only in tumoral mitochondria, and not in mitochondria obtained from extra-tumoral tissue (P<0.05). Conclusion: In conclusion this study suggested that fraction 2 of Cassiopea andromeda crude venom selectively induces ROS mediated cytotoxicity by directly targeting mitochondria isolated from human breast tumor tissue.
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BACKGROUND: The aim of present work was to assess the concentration levels as well as vertical distribution of indicator bacteria including total coliform, fecal coliform, Pseudomonas aeruginosa, and Heterotrophic Plate Count (HPC) in the marine environment (seawater and coastal sediments) and evaluate the correlation between indicator bacteria and some physicochemical parameters of surface sediments as well as seawaters. METHODS: A total number of 48 seawater and sediment samples were taken from 8 stations (each site 6 times with an interval time of 2 weeks) between June and September 2014. Seawater and sediment samples were collected from 30 cm under the surface samples and different sediment depths (0, 4, 7, 10, 15, and 20 cm) respectively, along the Persian Gulf in Bushehr coastal areas. RESULTS: Based on the results, the average numbers of bacterial indicators including total coliform, fecal coliform, and Pseudomonas aeruginosa as well as HPC in seawater samples were 1238.13, 150.87, 8.22 MPN/100 ml and 1742.91 CFU/ml, respectively, and in sediment samples at different depths (from 0-20 cm) varied between 25 × 103 to 51.67 × 103, 5.63 × 103 to 12.46 × 103, 17.33 to 65 MPN/100 ml, 36 × 103 to 147.5 × 103 CFU/ml, respectively. There were no statistically significant relationships between the indicator organism concentration levels with temperature as well as pH value of seawater. A reverse correlation was found between the level of indicator bacteria and salinity of seawater samples. Also results revealed that the sediment texture influenced abundance of indicators bacteria in sediments. As the concentration levels of indicators bacteria were higher in muddy sediments compare with sandy ones. CONCLUSION: Result conducted Bushehr coastal sediments constitute a reservoir of indicator bacteria, therefore, whole of the indicators determined were distinguished to be present in higher levels in sediments than in the overlying seawater. It was concluded that the concentration levels of microbial indicators decreased with depth in sediments. Except total coliform, the numbers of other bacteria including fecal coliform, Pseudomonas aeruginosa and HPC bacteria significantly declined in the depth between 10 and 15 cm.
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BACKGROUND: The present work was designed to detect heavy metal contents of Al, Zn, Fe, Mn, Ni, V, Co, Cr and Cu in sediments and shells of the Trachycardium lacunosum collected in polluted and unpolluted areas along the Persian Gulf. METHODS: The samples were taken from surface sediments (0-10 cm) and shells of Trachycardium lacunosum in two separated areas (polluted and unpolluted) in northern part of the Persian Gulf, Asaluyeh Bay, during summer 2013. The prepared samples were analyzed by inductively coupled plasma-optical emission spectrometry (ICP-OES). RESULTS: Based on the results, all measured metals including Al, Zn, Fe, Mn, Ni, V, Co, Cr and Cu were meaningfully higher in the sediment samples of polluted area compared to unpolluted area and the order of metal concentrations in the sediment samples were Cr > Co > V > Ni > Zn > Cu > Fe > Al > Mn in polluted area. In the case of shell samples of Trachycardium lacunosum, polluted area contained significantly higher contents of Al, Zn, Fe, Mn, Ni, Co, Cr and Cu compared to unpolluted area and the order of metal concentrations in the shell samples were Fe > Zn > Al > Mn > Cu > Cr > Ni > Co in the polluted area. CONCLUSION: It was concluded that shells of the Trachycardium lacunosum can be used as a suitable bioindicator for heavy metals in the aquatic environment. Results confirmed that due to the possible contaminations by oil and gas activities near the polluted area perennial monitoring and mitigation measures is extremely necessary.
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Abstract Natural compounds from marine organisms have been rarely studied for their acetylcholinesterase inhibitory activities. The aim of this study was to isolate novel compounds with antiAChE activity from the venom of upside-down jellyfish Cassiopea andromeda Forskål, 1775. The compounds of the fractionated venom on gel filtration chromatography were identified by analyzing gas chromatography-mass spectroscopy data. The structure of the isolated compound that showed the most potent antiAChE activity in a docking study was elucidated by different spectral data, including 1H NMR and 13C NMR. Three compounds, including a neurosteroidal alkaloid androtoxin B, were identified from two venom fractions. This neurosteroidal alkaloid showed strong acetylcholinesterase inhibitory activity (IC50 2.24 ± 0.1 µM) compared with the reference standard, galantamine. The results obtained by a docking study demonstrated that Androtoxin B had close contact with two of the three amino acid residues of the catalytic triad of acetylcholinesterase gorge and was accommodated within a peripheral hydrophobic pocket composed of numerous aromatic site chains. In conclusion, the isolated neurosteroidal alkaloid from Cassiopea andromeda was a potent antiAChE agent with strong binding to both the catalytic and peripheral sites of acetylcholinesterase that correlated well with the experimental data. Further studies are required to determine whether androtoxin B could be a potential treatment for Alzheimer's disease.