Anticancer Effects of New Ceramides Isolated from the Red Sea Red Algae Hypnea musciformis in a Model of Ehrlich Ascites Carcinoma: LC-HRMS Analysis Profile and Molecular Modeling

Different classes of phytochemicals were previously isolated from the Red Sea algae Hypnea musciformis as sterols, ketosteroids, fatty acids, and terpenoids. Herein, we report the isolation of three fatty acids—docosanoic acid 4, hexadecenoic acid 5, and alpha hydroxy octadecanoic acid 6—as well as three ceramides—A (1), B (2), and C (3)—with 9-methyl-sphinga-4,8-dienes and phytosphingosine bases. Additionally, different phytochemicals were determined using the liquid chromatography coupled with electrospray ionization high-resolution mass spectrometry (LC-ESI-HRMS) technique. Ceramides A (1) and B (2) exhibited promising in vitro cytotoxic activity against the human breast adenocarcinoma (MCF-7) cell line when compared with doxorubicin as a positive control. Further in vivo study and biochemical estimation in a mouse model of Ehrlich ascites carcinoma (EAC) revealed that both ceramides A (1) and B (2) at doses of 1 and 2 mg/kg, respectively, significantly decreased the tumor size in mice inoculated with EAC cells. The higher dose (2 mg/kg) of ceramide B (2) particularly expressed the most pronounced decrease in serum levels of vascular endothelial growth factor -B (VEGF-B) and tumor necrosis factor-α (TNF-α) markers, as well as the expression levels of the growth factor midkine in tumor tissue relative to the EAC control group. The highest expression of apoptotic factors, p53, Bax, and caspase 3 was observed in the same group that received 2 mg/kg of ceramide B (2). Molecular docking simulations suggested that ceramides A (1) and B (2) could bind in the deep grove between the H2 helix and the Ser240-P250 loop of p53, preventing its interaction with MDM2 and leading to its accumulation. In conclusion, this study reports the cytotoxic, apoptotic, and antiangiogenic effects of ceramides isolated from the Red Sea algae Hypnea musciformis in an experimental model of EAC.

Culture of Three Seaweed Species in Cox’s Bazar Coast, Bangladesh

The experimental culture of 3 seaweed species, i.e. Hypnea musciformis, Enteromorpha intestinalis and Padina tetrastromatica in three locations of Cox’s Bazar coast, Bangladesh using 4×4 m horizontal coir rope net method were conducted during 2016 to 2018. Results showed that the biomass yield was highest from H. musciformis (30.23±0.40 kg fw m−1) and significantly differ from E. intestinalis (24.50±0.08 kg fw m−1) and P. tetrastromatica (10.18±0.45 kg fw m−1). The Daily Growth Rate (DGR) of H. musciformis (8.88% day−1) was also higher than the DGR of E. intestinalis (6.55% day−1) and P. tetrastromatica (6.74% day−1). Saint Martin’s Island showed higher biomass yield of seaweeds than Bakkhali and Inani due to favourable water quality. In context to physico-chemical parameters of seaweed farming, Cox's Bazar coast is suitable for seaweed cultivation, although more suitable culture areas and longer culture period yet to be identified. The findings of the present study suggest that the cultivation of H. musciformis is viable and coastal people can take up such activities as seasonal income generating activity in coastal water. Bangladesh J. Zool. 49(1): 47-56, 2021

Inclusion of Red seaweed (Hypnea musciformis) improves nutritional quality of vegetable salad

Nutritional properties of vegetable salad incorporated with red seaweed (Hypnea musciformis) wasevaluated. Two lots of vegetable salad were prepared viz., control salad without adding seaweed and seaweedsalad with inclusion of 20% seaweed along with cucumber, tomato, carrot, cloves, garlic, lemon and salt.Micronutrient composition indicated that addition of seaweed increased the Ca, Na, Fe and Zn content ofvegetable salad. Calcium and iron content was substantially higher in seaweed mixed salad. The resultsindicate that incorporation of seaweed in vegetable salad improves the nutritional content and hence can beused to enrich nutritional quality of food salad.

Enzymes associated with anti-inflammatory potentialities of purified terpenoid extracts from the selected sea weeds

Introduction: Macrophages are phagocytic WBCs involved in the immune defense and will be activated during inflammatory disorders. The synthesis of cytokines and mediators, particularly nitric oxide (NO) is triggered by the macrophage activator. NO induces many biological events. Therefore, NO regulation is proven to be potential for exploring anti-inflammatory drugs. Aim: The anti-inflammatory action of the purified terpenoid extracts from red algae such as Hypnea musciformis, Gracilaria dura and Kappaphycus alvarezii on LPS induced RAW 264.7 macrophages on lipoxygenase, cyclooxygenase, hyaluronidase, xanthine oxidase and myeloperoxidase inhibitory effects were evaluated. Methods: The methanolic extract of the sea weeds were subjected to silica gel column chromatography and the fraction was further subjected to GC-MS analysis. Then, the potentiality of the purified terpenoid extracts to inhibit various inflammation causing enzymes such as COX, LOX, hyaluronidase, xanthineoxidase and myeloperoxidase were carried out. Findings: The terpenoid extracts reduced the enzyme activities in a dose dependent manner as compared to control group. The extracts inhibited xanthine oxidase activity effectively at 250 µg/ml i.e., a maximum inhibitory activity of 62.1% as compared to the standard drug, allopurinol. The extract significantly inhibited lipoxygenase activity, with highest inhibitory activity at 100 µg/ml. The nitric acid synthesis was reduced to 8.5 µM by Hypnea musciformis. Conclusion: The present study revealed that the purified terpenoid extracts from H. musciformis exhibited potent anti-inflammatory activities followed by G. dura and K. alvarezii via regulating the anti-inflammatory enzymes. These findings provide justification for the traditional use of the red algae in inflammatory conditions.

Comparison of Anti-inflammatory, Analgesic, Antipyretic Properties of Selected Red algae - Hypnea musciformis (Wulfen) J V Lamouroux., Gracilaria dura (C Agardh) J Agardh. and Kappaphycus alvarezii (Doty) Doty ex Silva

Marine natural products from seaweeds have been the focus of research for novel products of pharmacological interest. Bioactivities of the phytochemicals derived from marine algae are important ingredients in many products, such as cosmetics and drugs for treating cancer and other lifestyle disorders. In this context, the aim of the present study is to compare the anti-inflammatory, analgesic, antipyretic properties of the purified terpenoids from the selected red algae Hypnea musciformis, Gracilaria dura and Kappaphycus alvarezii. Initially, the acute toxicity of the purified terpenoid extract was analyzed and was greater than 4000 mg/kg bw. In vitro BSA denaturation assay revealed significant % inhibition as compared to control. In the carrageenan induced paw edema, the purified terpenoids extract of H. musciformis exhibited remarkable soothing activity with 68.8% percentage of inhibition, which was comparable with that of indomethacin. Meanwhile G. dura and K. alvarezii showed 53 and 50% of inhibition respectively. Analgesic activity was determined by hot plate and acetic acid induced writhing test. In the hot plate test, terpenoids extract of H. musciformis and G. dura significantly increased the hot-plate latency as compared to normal saline, which reflects their analgesic efficacy. In the writhing test, terpenoid extracts from H. musciformis, G. dura and K. alvarezii inhibited the writhing response induced by acetic acid in a concentration dependent manner that suggests its varied mode of inhibition of stretching episodes. Lastly, the antipyretic activity was analyzed. Terpenoid extracts from H. musciformis, G. dura and K. alvarezii revealed a significant (P < 0.01) antipyretic activity up to 180 min. In summary, the study demonstrates the anti-inflammatory, analgesic and antipyretic effects of terpenoids extracts from H. musciformis, G. dura and K. alvarezii on experimental models, suggesting its therapeutic potential in the treatment of peripheral painful and inflammatory pathologies.

Screening of in vitro antiviral activity of purified terpenoid extracts of selected sea weeds against chikungunya virus

Currently, the search of novel phytochemicals with unique biological potentialities is a pre-requisite for the designing ideal drugs for the human kind. Sea weeds are bioresources with a broad spectrum of medicinal properties with minimal side effects. Kerala, the Southern state of India reported high incidence of Chikungunya virus (CHIKV) infections in the last several tears. No specific virucidal therapy or effective vaccines are available. This emphasizes the need of searching for phytochemicals as drugs with less cost and more effective. Therefore, an attempt was made in screening purified terpenoid extracts of selected sea weeds as anti-CHIKV potential. In this study the terpenoids composition from the red algae Hypnea musciformis, Kappaphycus alvarezii and Gracillaria dura were identified and analyzed by thin layer chromatography and Gas chromatography- Mass spectrum. The methanolic extract of seaweeds was purified by column chromatography and each fraction was eluted by using petroleum ether and ethyl acetate as solvent combination. The analysis of the purified fraction of H. musciformis and K. alvarezii revealed the presence of 8 terpenoid fractions, and G. dura showed only 4 major components respectively. Vero cell lines were employed in the antiviral assays, infected to CHIKV, and treated with varied doses of purified terpenoid extracts. In the antiviral activity, terpenoid extracts of G. dura showed remarkable and promising EC50 inhibitory effect at 1.25 μg/ml. Further, the terpenoid extracts displayed efficient virucidal activity against CHIKV (inhibit around 90%) with 5 μg/ml dosage. As the last phase, terpenoid extracts added at time intervals of 0, 1, 2, 3 post-infection periods still maintained a significant inhibitory potential against CHIKV viral replication. Thus, the overall study suggests that the terpenoid extracts of G. dura may be effectively used in the prevention and treatment of CHIKV infections. Clinical studies may be warranted for designing a promising new anti-CHIKV drug.

Chemical Biopreservative Effects of Red Seaweed on the Shelf Life of Black Tiger Shrimp (Penaeus monodon)

Hypnea musciformis (HM) and Acanthophora muscoides (AM) red seaweeds were evaluated for their antioxidant properties and efficacy to extend the chemical shelf life of black tiger shrimp (Penaeus monodon) during 14-daystorage. Treated shrimp were soaked in five percent ethanolic solution with 500 µg/mL of AM or HM powder for 30 min. HM had more phenols and flavonoids, increased radical scavenging activity, and greater H2O2 reducing power than AM in vitro. Biochemical quality indicators were significantly higher in the control group, followed by HM- and AM-treated samples during storage. On day 14 of storage, controls contained significantly higher amounts of biogenic amines than HM- or AM-treated samples. The shelf life of chilled stored shrimp increased due to the presence of compounds of butylated hydroxytoluene, sulfurous acid, heptadecane, mono (2-ethylhexyl), and 1,2-propanediol found in AM extract and sulfurous acid and 1,2-propanediol found in HM extract. A control group was soaked in the same ethanolic solution as treated samples without algae powder for 30 min. Each group was kept ice-cold during the soaking period. The results obtained demonstrate the usefulness of two seaweed extracts, Hypnea musciformis and Acanthophora muscoides, combined with ice by decreasing the formation of toxic biogenic amines in shrimp, enhancing its shelf life during ice storage.