Evaluation of disease resistance and tolerance to elevated temperature stress of the selected tissue-cultured Kappaphycus alvarezii Doty 1985 under optimized laboratory conditions

Information on the abiotic stress tolerance and ice–ice disease resistance properties of tissue-cultured Kappaphycus alvarezii is scarce and can pose a big hurdle to a wider use of tissue-cultured seaweed in the industry. Here, we reported on a study of seaweed-associated bacteria diversity in farmed and tissue-cultured K. alvarezii, and ice–ice disease resistance and elevated growth temperature tolerance of tissue-cultured K. alvarezii in laboratory conditions. A total of 40 endophytic seaweed-associated bacteria strains were isolated from 4 types of K. alvarezii samples based on their colony morphologies, Gram staining properties and 16S rRNA gene sequences. Bacteria strains isolated were found to belong to Alteromonas sp., Aestuariibacter sp., Idiomarina sp., Jejuia sp., Halomonas sp., Primorskyibacter sp., Pseudoalteromonas sp., Ruegeria sp., Terasakiella sp., Thalassospira sp. and Vibrio sp. Vibrio alginolyticus strain ABI-TU15 isolated in this study showed agar-degrading property when analyzed using agar depression assay. Disease resistance assay was performed by infecting healthy K. alvarezii with 105 cells/mL Vibrio sp. ABI-TU15. Severe ice–ice disease symptoms were detected in farmed seaweeds compared to the tissue-cultured K. alvarezii. Besides disease resistance, tissue-cultured K. alvarezii showed better tolerance to the elevated growth temperatures of 30 and 35 °C. In conclusion, our overall data suggests that tissue-cultured K. alvarezii exhibited better growth performance than farmed seaweeds when exposed to elevated growth temperature and ice–ice disease-causing agent

Interleukin-27 disrupts the crosstalk of apoptotic activities between 4T1 breast cancer cells and M2 macrophages

Introduction: Cytokine immunotherapy such as Interleukin-27 (IL-27) has been foreseen as a promising alternative anti-cancer treatment. Thus, this study aimed to investigate whether IL-27 gene therapy regulates crosstalk between breast cancer cells and macrophages in the sense of pro-apoptotic activities. Methods: This study has led to the development of recombinant pcDNA3.4-IL27. The recombinant pcDNA3.4-IL27 was transfected into 4T1 murine mammary carcinoma cells alone and co-culture of 4T1 with M2 macrophages. The successful expression of IL-27 in the cells were determine through the immunofluorescence staining and detection of CD206, M2 macrophages marker. Apoptotic effects of pcDNA3.4-IL27 were assessed through MTT assay, Annexin V flow cytometer analysis, and AO/PI dual staining. Results: Our findings shows that pcDNA3.4-IL27 has the ability to induce apoptosis in both of the cell group and performs better in the co-culture of 4T1 with M2 macrophages compared to 4T1 cells alone. PcDNA3.4-IL27 induced apoptosis through the altered cell morphology and reduction in the number of viable cells. Conclusion: These data demonstrate that pcDNA3.4-IL27 has the ability to induce apoptosis in both 4T1 cell alone and co-cultured 4T1 with M2 macrophages. Thus, could serve as a potential anti cancer candidate against breast cancer

Molecular investigation of carrageenan production in Kappaphycus alvarezii in different culture conditions: a proteomic approach

Carrageenan is a polysaccharide extracted from different species of red seaweed for use in pharmaceuticals and cosmetics industries. In Malaysia, κ-carrageenan is obtained through extraction from the tropical red seaweed Kappaphycus alvarezii. The use of tissue culture techniques in the propagation of K. alvarezii has proven to be effective in solving cultivation problems and produced high-quality seedlings and providing a sustainable source of better quality carrageenan. In order to understand the molecular mechanisms behind that, a proteomic investigation was conducted comparing the changes in protein expression in tissue-cultured and liquid-cultured K. alvarezii after 60 days of cultivation. Proteomic analysis was used to study the changes in the protein expression level between liquid-cultured and tissue-cultured of the red seaweed K. alvarezii. A total of 45 protein spots were found to be significantly different in their densities and three proteins, namely β-amylase, NAD-dependent sugar epimerase and B-phycoerythrin, showed a consistent pattern of upregulation in ELISA analyses, hence validating the 2-DE profiles. Changes in the proteins expression level were noticed in proteins related to energy production, metabolism and cellular maintenance. The protein changes in tissue-cultured seaweed possibly play an essential role in the production of carrageenan