In-vitro and in-silico anti-HSV-1 activity of a marine steroid from the jellyfish Cassiopea andromeda venom.

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.

The Effect of the Persian Gulf Jellyfish (Cassiopea andromeda) Venom on the Expression of P15, P21, P53, DNMT1, and Bcl-2 in Acute Lymphoblastic Leukemia Jurkat Cells.

Background: One of the acute hematologic malignancies is acute lymphoblastic leukemia (ALL), which is formed in B or T lymphocyte stem cells. Regarding the increasing tendency to herbal and marine studies and unclear characteristics of Cassiopea andromeda Venom, this study was performed to determine its effects on Jurkat cells as a model for T-ALL. Materials and Methods: In this experimental study, the cells were treated with a variety of concentrations of Cassiopea andromeda venom at different periods and times. Growth inhibition and toxic effects of Cassiopea andromeda Venom were evaluated by methyl thiazole tetrazolium salt reduction (MTT test). The flow cytometry analysis was carried out using 7-aminoactinomycin D (7AAD) and Annexin V stains to evaluate the venom's effect on apoptotic pathways. Besides, Real-Time PCR was performed to evaluate the relative gene expression. Results: Cassiopea andromeda venom inhibited the growth of Jurkat cells in a concentration and time manner. Jurkat cell growth was inhibited by 48.9% after 72 hours of treatment with 250µg/mL Cassiopea andromeda venom. The venom increased the apoptotic process through the upregulation of p15INK4b and P53 proteins and downregulation of Bcl-2, p21 WAF1/CIP1, and DNMT1 in the Jurkat cell line. Conclusion: Considering the growth inhibitory property of Cassiopea andromeda Venom, we recommend it as a part of combinational medication for treating ALL in animal trials and for other leukemias in vitro studies.

Tissue Extract from Brittle Star Undergoing Arm Regeneration Promotes Wound Healing in Rat.

This study set out to evaluate the wound healing properties of brittle star extracts in vitro and in vivo. Due to the great arm regeneration potential of the brittle star, Ophiocoma cynthiae, the present study aimed to evaluate the wound healing effect of hydroalcoholic extracts of brittle star undergoing arm regeneration in wound healing models. The brittle star samples were collected from Nayband Bay, Bushehr, Iran. After wound induction in the arm of brittle stars, hydroalcoholic extracts relating to different times of arm regeneration were prepared. The GC-MS analysis, in vitro MTT cell viability and cell migration, Western blot, and computational analysis tests were performed. Based on the in vitro findings, two BSEs were chosen for in vivo testing. Macroscopic, histopathological and biochemical evaluations were performed after treatments. The results showed positive proliferative effects of BSEs. Specifically, forty-two compounds were detected in all groups of BSEs using GC-MS analysis, and their biological activities were assessed. The MTT assay showed that the 14 d BSE had a higher proliferative effect on HFF cells than 7 d BSE. The cell migration assay showed that the wound area in 7 d and 14 d BSEs was significantly lower than in the control group. Western blot analysis demonstrated an increase in the expression of proliferation-related proteins. Upon the computational analysis, a strong affinity of some compounds with proteins was observed. The in vivo analysis showed that the evaluation of wound changes and the percentage of wound healing in cell migration assay in the 7 d BSE group was better than in the other groups. Histopathological scores of the 7 d BSE and 14 d BSE groups were significantly higher than in the other groups. In conclusion, the hydroalcoholic extract of O. cynthiae undergoing arm regeneration after 7 and 14 days promoted the wound healing process in the cell and rat skin wound healing model due to their proliferative and migratory biological activity.

Proliferative Effect of Aqueous Extract of Sea Cucumber (Holothuria parva) Body Wall on Human Umbilical Cord Mesenchymal Stromal/Stem Cells.

Sea cucumber extracts and their bioactive compounds have the potential for stem cell proliferation induction and for their beneficial therapeutic properties. In this study, human umbilical cord mesenchymal stromal/stem cells (hUC-MSCs) were exposed to an aqueous extract of Holothuria parva body walls. Proliferative molecules were detected using gas chromatography-mass spectrometry (GC-MS) analysis in an aqueous extract of H. parva. The aqueous extract concentrations of 5, 10, 20, 40, and 80 µg/mL and 10 and 20 ng/mL of human epidermal growth factor (EGF) as positive controls were treated on hUC-MSCs. MTT, cell count, viability, and cell cycle assays were performed. Using Western blot analysis, the effects of extracts of H. parva and EGF on cell proliferation markers were detected. Computational modeling was done to detect effective proliferative compounds in the aqueous extract of H. parva. A MTT assay showed that the 10, 20, and 40 µg/mL aqueous extract of H. parva had a proliferative effect on hUC-MSCs. The cell count, which was treated with a 20 µg/mL concentration, increased faster and higher than the control group (p < 0.05). This concentration of the extract did not have a significant effect on hUC-MSCs' viability. The cell cycle assay of hUC-MSCs showed that the percentage of cells in the G2 stage of the extract was biologically higher than the control group. Expression of cyclin D1, cyclin D3, cyclin E, HIF-1α, and TERT was increased compared with the control group. Moreover, expression of p21 and PCNA decreased after treating hUC-MSCs with the extract. However, CDC-2/cdk-1 and ERK1/2 had almost the same expression as the control group. The expression of CDK-4 and CDK-6 decreased after treatment. Between the detected compounds, 1-methyl-4-(1-methyl phenyl)-benzene showed better affinity to CDK-4 and p21 than tetradecanoic acid. The H. parva aqueous extract showed proliferative potential on hUC-MSCs.

Chemical Compositions and Experimental and Computational Modeling of the Anticancer Effects of Cnidocyte Venoms of Jellyfish Cassiopea andromeda and Catostylus mosaicus on Human Adenocarcinoma A549 Cells.

Nowadays, major attention is being paid to curing different types of cancers and is focused on natural resources, including oceans and marine environments. Jellyfish are marine animals with the ability to utilize their venom in order to both feed and defend. Prior studies have displayed the anticancer capabilities of various jellyfish. Hence, we examined the anticancer features of the venom of Cassiopea andromeda and Catostylus mosaicus in an in vitro situation against the human pulmonary adenocarcinoma (A549) cancer cell line. The MTT assay demonstrated that both mentioned venoms have anti-tumoral ability in a dose-dependent manner. Western blot analysis proved that both venoms can increase some pro-apoptotic factors and reduce some anti-apoptotic molecules that lead to the inducing of apoptosis in A549 cells. GC/MS analysis demonstrated some compounds with biological effects, including anti-inflammatory, antioxidant and anti-cancer activities. Molecular docking and molecular dynamic showed the best position of each biologically active component on the different death receptors, which are involved in the process of apoptosis in A549 cells. Ultimately, this study has proven that both venoms of C. andromeda and C. mosaicus have the capability to suppress A549 cells in an in vitro condition and they might be utilized in order to design and develop brand new anticancer agents in the near future.

MRI Tracking of Marine Proliferating Cells In Vivo Using Anti-Oct4 Antibody-Conjugated Iron Nanoparticles for Precision in Regenerative Medicine.

Marine invertebrates are multicellular organisms consisting of a wide range of marine environmental species. Unlike vertebrates, including humans, one of the challenges in identifying and tracking invertebrate stem cells is the lack of a specific marker. Labeling stem cells with magnetic particles provides a non-invasive, in vivo tracking method using MRI. This study suggests antibody-conjugated iron nanoparticles (NPs), which are detectable with MRI for in vivo tracking, to detect stem cell proliferation using the Oct4 receptor as a marker of stem cells. In the first phase, iron NPs were fabricated, and their successful synthesis was confirmed using FTIR spectroscopy. Next, the Alexa Fluor anti-Oct4 antibody was conjugated with as-synthesized NPs. Their affinity to the cell surface marker in fresh and saltwater conditions was confirmed using two types of cells, murine mesenchymal stromal/stem cell culture and sea anemone stem cells. For this purpose, 106 cells of each type were exposed to NP-conjugated antibodies and their affinity to antibodies was confirmed by an epi-fluorescent microscope. The presence of iron-NPs imaged with the light microscope was confirmed by iron staining using Prussian blue stain. Next, anti-Oct4 antibodies conjugated with iron NPs were injected into a brittle star, and proliferating cells were tracked by MRI. To sum up, anti-Oct4 antibodies conjugated with iron NPs not only have the potential for identifying proliferating stem cells in different cell culture conditions of sea anemone and mouse cell cultures but also has the potential to be used for in vivo MRI tracking of marine proliferating cells.

Toxic effect of cigarette butts leachates on blood markers of Periophthalmus waltoni species from the Persian Gulf region.

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.

Chemical contents and toxicity of cigarette butts leachates in aquatic environment: A case study from the Persian Gulf region.

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.

Antioxidant and anticholinesterase properties of Echinometra mathaei and Ophiocoma erinaceus venoms from the Persian Gulf.

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.

Metal uptake in Psettodes erumei and Hysterothylacium spp. larvae in the Persian Gulf: Evaluation of larvae as bio-indicator.

This study aimed to compare the concentrations of heavy metals in Psettodes erumei as host fish and larvae of Hysterothylacium spp. as its parasite. Moreover, to evaluate the larvae as bio-indicators the uptake of heavy metals, the infected and non-infected fish were also compared. Fresh P. erumei species (n = 19) were randomly sampled during four months from Bushehr County, Iran. The digestive tract of each fish was examined for nematode parasites using a stereomicroscopy. The isolated nematodes were identified, and content of Fe, Cu, Zn, Co, Ni, Cr, As, Cd, Hg, and Pb were measured using ICP-OES. The metal concentrations were simultaneously analyzed for the muscles of infected fish and their parasites, as well as non-infected ones. Of the 19 P. erumei examined, 13 (68.4%) P. erumei were infected with Hysterothylacium spp. larvae. The parasites had significantly higher level of Fe, Cu, Zn, and Ni (with mean value of 7.59, 0.572, 1.223, and 4.623 mg/kg, respectively) than the muscles of the host fishes (with mean value of 3.29, 0.0010, 0.586, and 0.277 mg/kg, respectively) (p < 0.05). Infected hosts showed significantly lower amounts of As element in their muscles (0.050 mg/kg) than non-infected hosts (0.113 mg/kg) (p < 0.05). The findings emphasize the potential role of Hysterothylacium spp. larvae as bio-indicators for monitoring heavy metals pollution in marine ecosystems.

Studies on the cholinesterases inhibiting compounds from the Cassiopea andromeda venom.

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.

Synthesis, Structural Studies, and α-Glucosidase Inhibitory, Antidiabetic, and Antioxidant Activities of 2,3-Dihydroquinazolin-4(1H)-ones Derived from Pyrazol-4-carbaldehyde and Anilines.

A series of new quinazoline derivatives were designed and synthesized via a one-pot condensation reaction between isatoic anhydride and aromatic aldehydes with anilines using aluminum sulfate as a catalyst in refluxing ethanol. Their structures were confirmed by their physical, IR, 1H NMR, 13C NMR, and mass spectroscopy data and evaluated for some biological effects, including the antioxidant and α-glucosidase inhibitory activities as well as some in vivo hematological parameters. The ability of synthesized compounds in the inhibition of α-glucosidase was also investigated through the in silico study. The significant and important changes in some hematological tests were perceived. Notably, compound 4h showed more reducing effects on cholesterol and triglyceride levels. This molecule certainly has the potential to be developed as the antihyperlipemic compound. The tested compounds, in particular, compounds 4j and 4l, were found to be uniquely reducing blood sugar levels. The entire synthesized compounds showed the potent α-glucosidase inhibitory activity compared with acarbose as a standard material. Amongst, the compounds 4h and 4i showed the strongest enzyme inhibitory potentials than the standard drug acarbose. There was a good correlation between in vitro and in silico studies for ligands 4i and 4l. The majority of compounds presented a good radical scavenging activity, though the compound 4j exhibited the strongest activity, even to the standard of ascorbic acid. Further studies are required to determine whether these main compounds could be a potential treatment for diabetes and hyperlipidemia diseases.

Antibacterial studies of hydroxyspiro[indoline-3,9-xanthene]trione against spiro[indoline3,9-xanthene]trione and their use as acetyl and butyrylcholinesterase inhibitors.

Xanthene derivatives are well known for their effective biological activities. In search of effective antibacterial agents, the spiro[indoline3,9-xanthene]-trione (A) and hydroxy-spiro[indoline-3,9-xanthene]-trione (B), were synthesized and tested for in vitro antibacterial activity against Staphylococcus aureus and Escherichia coli. Furthermore, the synthesized compounds were tested in vitro and in silico for their anticholinesterase activities. The anticholinesterase activities for six substitutes of the hydroxy derivative (B1-B6) were also studied through the molecular docking. All concentrations of compounds presented a dose-dependent antibacterial activity. The docking results showed that all compounds are more constant than the galantamine. Amongst, compound B1 exhibited the minimum binding energy in both AChE and BChE enzymes. Results indicate the importance of xanthene derivatives as potential antibacterial and anticholinesterases agents.

Healing effects of Pergularia tomentosa L., a native medicinal plant in Bushehr province, Iran on burn, in animal model.

Burn injury is one of the most destructive events in the world. The Pergularia tomentosa L. is a medicinal plant that traditionally, applies for treatment of burning, in Bushehr province, Iran. Various bioactive compounds such as steroid glycosides, tannins, various vitamins, saponins, cardenolides and anthraquinones were identified into extract of the plant, which can be effective in burn wound healing. Twenty-one rats weighting every one 200±5 grams were divided equally into three groups. The second-degree burning induced on all groups. One of groups did not receive any treatment (The control group) and was treated locally with saline and eucerin. The Second group received the P. tomentosa L. as a topical ointment, and the third group received locally, a thin layer of silver sulfadiazine ointment 3% after washing the wound with saline. Afterward treatment period, the microscopic slides from histological sections were prepared. At that point, amounts of the fibroblast cells, blood vessels, wound area, necrotic tissues, and diameter of epidermis rate of wound healing were determined. Also the exterior status of wound in different days was considered. Results obtained from current study have revealed that the extract of P. tomentosa L. can significantly, cause qualitative and quantitative acceleration in healing of second degree burn wounds, due to their bioactive and vasoactive properties. In conclusion the P. tomentosa L. can is used as an overborne medicine with lower cost and side effect than the similar chemical medicines. Although, the further studies are needed on these plants, due to their some toxic effects.

Acetylcholinesterase inhibitory activity of a neurosteroidal alkaloid from the upside-down jellyfish Cassiopea andromeda venom

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.

Green synthesis of novel spiro-indenoquinoxaline derivatives and their cholinesterases inhibition activity.

A convenient synthesis of substituted spiroindenoquinoxalines at mild and green conditions was developed. Multicomponent reaction of substituted phenylene diamines, ninhydrin, malononitrile and N,N'-substituted-2-nitroethene-1,1-diamines produced the target compounds. Twelve new spiroindenoquinoxalines were obtained, and their ability in inhibition of acetyl and butyrylcholinesterases were investigated both in vitro and in silico. All compounds showed moderate level activity against both acetyl and butyrylcholinesterases.

Hematological parameters on the effect of the jellyfish venom Cassiopea andromeda in animal models.

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.

Application of PEG-400 as a green biodegradable polymeric medium for the catalyst-free synthesis of spiro-dihydropyridines and their use as acetyl and butyrylcholinesterase inhibitors.

A simple, efficient and green approach for the synthesis of spiro-dihydropyridines derivatives by one-pot multi-component reaction of isatin or acenaphthoquinone derivatives (1 equiv) with malononitrile (1 equiv) and N,N'-substituted-2-nitroethene-1,1-diamines (1 equiv) in PEG-400 under catalyst-free conditions is described. This method provides several advantages such as environmental friendliness, short reaction time, and simple workup procedure for the synthesis of biologically important compounds. The ability of synthesized compounds in inhibition of acetyl and butyrylcholinesterase were investigated both in vitro and in silico. All compounds showed moderate to high level activity against both acetyl and butyrylcholinesterase. There was a good correlation between in vitro and in silico studies.