The paper by Yong Chang Seo et al., report the effect of Spirulina maxima extracts on proliferation of various cell lines and cytokine production. The authors aim to demonstrate that growing S. maxima in deep sea water rather than sea water will enhance the effects of the extract. Besides the fact that the collection of deep sea water is complicated (and inaccessible to most) and that its composition may vary depending on the season, location, etc, the results presented do not seem meaningful. In addition, this paper suffers from several defects and some of the conclusions are not really supported by the presented results. Major general concerns The results presented are disparate and lack a common thread. Different results were obtained in different cell lines and it would have been much more appropriate to study all the parameters in all the appropriate cell lines. How could the extracts act as proliferation enhancer of Raji and Jurkat cell lines and have antiproliferative effect on HEK293, AGS, MCF-7 and Hep3B cells within the same range of concentration? And what about A549 cells? A clear presentation of the proliferation effect of the extract on all cell lines used in the same Figure should be provide instead of the disparate Figures and Tables. The Bcl2 expression should be investigated in more than one cell line (especially when it was not used in proliferation assays). See also specific concern below. But this downregulation of Bcl2 should not be over-interpreted and is not sufficient to cause carcinogenesis. In addition, results should be confirmed by using pure molecules supposed to be relevant among those present in the extracts. Most of them are available including ascorbic acid, carotenes, PUFAs… This may lead to the identification of bioactive molecules or highlight the synergy between them in the S. maxima extract. Major specific concerns L 28 In the abstract: “indicating that the extracts may play more important roles in inhibiting the initiation step of carcinogenesis”, the authors are not studying carcinogenesis and Bcl2 downregulation per se is not sufficient to talk about carcinogenesis. L 63 The “pathogen-free” status of deep sea water remains to be proven. The best no known pathogen has been found but what about unknown? And this could change… I noted that the authors were a little more cautious at the end of the manuscript (L 327). L 90 Table 2 titles “several cancer cell lines” but only 2 were used! Fig 3 The quality of this photograph should be enhanced and a wider view of the western blot result must be provided. A loading control must be added consisting of either housekeeping protein immunodetection or (at least) coomassie gel staining. Fig 5 The quality of this figure should be greatly enhanced. In its present form it was not possible to evaluate it accurately. Minor concerns L 26/27 Please use Bcl2 as the name of the gene/protein instead of “B cell lymphoma-2” or at least add the term “gene” or “protein” behind. L 43 “Many studies…” and you cite only 3 !!! L 57 S. maxima L 74 Please define here what you mean by “selectivity”. This come in the methods section but far later. L 102 W-SW instead of W-DSW? L 246 Table 4 (instead of Table 1) L 249 What are solution A5 and solution B6? It should be indicated. L 250 I do not understand what mean “two filtered basal seawater like a conventional SW or DSW. I think some words are missing. Please explain. L 278 3.4 title: “Measurement of Bcl2 protein level” L 290 3.5 title: “Measurement of human T and B…” L 327 Should be cautious with the “relatively pathogen free status” of deep sea water (see above)
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