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)
International Journal of Molecular Sciences
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The authors report about the preparation and characterization of an experimental dental restorative composite, which should be promising thanks to its radio opaque and antibacterial actions. Indeed a lot of valuable experimental work has been done and is presented. However, most of it seems not to be novel, as mentioned by the authors themselves. First, the properties of DMMAI itself are well known. Second, the authors themselves have already been deeply investigating the same or little different material. By the way, the authors are extensively self citing (Refs. 24, 32, 39, 47, 55, 65, 66), which is too much (7 occurrences) to be believed to be really necessary to present the background of the work. In fact, the total number of references (66) is also too high, probably made to justify the number of above self citations. Back to the work contents, the novelty of the present composite formulation would be the use of DDMAI (imparting both properties of interest) to BisGMA MMA instead of BisGMA TEGDMA (as in Ref.32), which did not allow to reach a DMMAI concentration higher than 5%, due to miscibility problems with TEGDMA, limiting the useful effects of DMMAI. In this work the authors replaced TEGDMA with MMA, and shifted the BisGMA comonomer relative concentration from 50:50 to 80:20. Thus, given the good mixing properties of the new formulation, they could reach up to 25% DDMAI to monomer. In my personal opinion, this is not sufficient in itself to justify a new research article, especially given the negative results. In fact, even if the DC is not affected by DDMAI, both the mechanical and aging properties (in water) clearly are, in negative way. Additionally to the decrease in these properties, absolute radio-opacity is also not sufficient. Only the antibacterial effect is really interesting. Therefore, probably this work could better fit a microbiological journal or one of those novel journals reporting about negative experimental results. The overall poor performance results are also evident in the actual weak form of section , Conclusion. Altogether, the work is not acceptable.
In the Results section, several considerations about the statistical significance of differences are not exact. In fact, even if the general flavor is correct, when speaking of statistical significance of their results the authors should be more precise. For example, they write that (page 4, lines 9-10) “control polymer had higher FS and FM than all polymers with DDMAI (p<0.05)”. Given the error bar plotted in Fig.3, this is probably false in the following cases: for FM, in both “before immersion”and “dried” condition, for control vs 15% DDMAI; for FS, in “dried” condition, for control vs 15%. Similarly, in lines 10 11: “FS and FM of polymers with DDMAI decreased with increase of DMMAI concentration (p<0.05)”, which should be false for: FS, before condition, for 15 vs 20%, and dried condition for 15 vs 20%; for FM, after condition, for 20 vs 25%. The trend is clear, but in these peculiar cases the difference should not be statistically effective.
Fig.4: an in-panel legend placing a distance scale by the side of the images themselves (line with arrow for thickness direction and values of levels, 0.5, 1, 1.5 etc. by the side) would surely help the reader to more easily interpret this figure.
Fig.5: all the right columns images should be removed, as this close up does not add any insight, and the larger view of the left column itself is sufficient to describe the presented effect. (BTW, this way the figure could be made a more compact 2x2 panels one, and no SEM would be justified but an optical microscope would suffice, always choose the easiest option).
Fig.6: because other peaks in the spectrum are changing even more than those two used to evaluate the DC (eg see around 1250 and 1050 cm^-1), either a zoomed region only is showed around the peaks of interest of an inset in top left could show a close up of that region.
Minor text edits follow (demonstrating thoroughful reading by the referee):
Page 2 line 40: “could not over”, change to “could not go over”.
Abstract, line 26: better use Degree of conversion before defining DC acronym, rather than Double bond conversion (which can make non expert reader think the D stands for double).
Page 8, line 2: “vitification”, change to “vitrification: wouldn’t be bad the paper authors always make their document go through a word processor chedckspeller, each and every time.
Page 8, line 31: it is not Fig.5 but is Fig.4.
Page 9, line 10: “into” change to “in”, no motion there.
Page 9 line 17 “days’” change to “days”.
Page 9 line 35:” used” change to “use”.
Page 10 line 24: “according” change to “according to”.
Page 11 line 7: “day” change to “ days”.
Line 9 (equation): “SL” to “WSL”.