On 2014 Nov 20, Daniel J Simons commented:

This paper reports data from the same training study that was previously reported in PLoS One: http://www.plosone.org/article/info:doi/10.1371/journal.pone.0092269#pone-0092269-g002. It reports results from different outcome measures, but it also reports the results from the earlier paper (including the same statistics and the same data figure: Figure 3b is nearly identical to Figure 2b in the PLoS paper). The paper does not acknowledge that these data were previously published. The training data and results also are presented in both places without acknowledgment of the overlap. The paper does not explicitly state that these are results from the same intervention that was published previously.

I have posted this comment on the Frontiers website as well, and I have written about these issues in more depth at http://blog.dansimons.com/2014/11/hi-bar-more-benefits-of-lumosity.html. I also wrote about problems with the PLoS paper in an earlier post-publication review (which is why I noticed the overlap): http://blog.dansimons.com/2014/04/hi-bar-benefits-of-lumosity-training.html. The Frontiers paper suffers from the same problems as the earlier paper, given that it was the same intervention.

On 2014 Nov 24, Guillaume Filion commented:

This article is one of the "CISCOM meta-analyses", which are very similar papers written by different authors. For more information about the CISCOM meta-analyses, check the blog post "A flurry of copycast on PubMed" at the following link http://blog.thegrandlocus.com/2014/10/a-flurry-of-copycats-on-pubmed

On 2015 Jun 18, Bill Cayley commented:

A great example of "less is more" in medicine: https://lessismoreebm.wordpress.com/2015/06/18/antibiotics-for-prelabour-rupture-of-membranes-at-or-near-term/

On 2016 Aug 22, Anthony Jorm commented:

The Royal Australian and New Zealand College of Psychiatrists (RANZCP) has recently published clinical practice guidelines on eating disorders Hay P, 2014, mood disorders Malhi GS, 2015 and schizophrenia Galletly C, 2016. These guidelines contain a mixture of evidence-based recommendations where there were relevant intervention studies, and consensus-based recommendations where relevant studies did not exist. The consensus-based recommendations comprised a substantial proportion of the guidelines for mood disorders (59%) and schizophrenia (46%), but less so for eating disorders (10%), indicating that expert consensus is an important source of guidance on best practice in psychiatry.

Given the substantial contribution of expert consensus to these guidelines, it is important that the methods for establishing this consensus are adequate. The Australian National Health and Medical Research Council (NHMRC) has published requirements for development of clinical practice guidelines, but these do not give much guidance on how this should be done, simply mandating that “The method used to arrive at consensus-based recommendations or points (e.g. voting or formal methods, such as Delphi) is documented”. (National Health and Medical Research Council. Procedures and requirements for meeting the 2011 NHMRC standard for clinical practice guidelines. Melbourne: National Health and Medical Research Council; 2011.)

Another potential source of criteria for evaluating the quality of methods for developing consensus-based recommendations comes from research on ‘wisdom of crowds’ Lorenz J, 2011 Kattan MW, 2016 Baumeister RF, 2016. Based on such research, Surowiecki has proposed four conditions necessary for a group to make good decisions (Surowiecki J. The wisdom of crowds: why the many are smarter than the few. London: Abacus; 2004.): 1. Diversity of expertise. A heterogeneous group of experts will produce better quality decisions than a homogeneous one. For guidelines developers, this may mean that the experts should come from a range of relevant disciplines, including consumer experts where appropriate. 2. Independence. The experts must be able to make their decisions independently, so that they are not influenced by others. For guidelines developers, this means that voting on consensus-based recommendations is carried out privately so that strong individuals cannot dominate the group. 3. Decentralization. Expertise is held by autonomous individuals working in a decentralized way. For guidelines developers, it is important to specify what sources of information the experts had available to them. 4. Aggregation. There is a mechanism for coordinating and aggregating the group’s expertise. For guideline developers, this could involve an independent person who runs the voting and gives feedback to the group.

If we take these four conditions as appropriate for judging the quality of methods for developing consensus-based recommendations, how well do the RANZCP guidelines meet them?

An indication of diversity of expertise is the disciplinary composition of the guideline working groups. There was limited diversity for all working groups, with non-psychiatrists comprising 3 of the 8 members for eating disorders, 4 out of the 12 members for mood disorders and 2 out of the 10 members for schizophrenia working group. There were no consumer or carer members of any of the working groups. While the mood disorder and schizophrenia guidelines included consensus-based recommendations s for indigenous peoples, it is not stated whether any of the working groups included indigenous members.

The mood disorders and schizophrenia working groups did not appear to involve independent decision making. Both groups had discussions until consensus was reached. The eating disorders guidelines did not give relevant information about whether there was independence.

All three guidelines state that consensus-based recommendations were based on collective clinical and research knowledge and experience. The eating disorder guidelines additionally state that level IV articles were considered where higher-level evidence was lacking and this informed the consensus-based recommendations.

After drafting, all guidelines had input from a broader group of expert advisers with a wide diversity of expertise. However, it is not clear whether these advisers had the potential to persuade working group members to change consensus-based recommendations.

Where the guidelines included consensus-based recommendations relevant to indigenous peoples, it is not clear what sources of cultural expertise these were based on.

None of the guidelines state how judgements were aggregated to determine consensus. The mood disorders guidelines state that agreement on consensus-based recommendations was “in most cases unanimous but allowed one committee member to abstain”. The other guidelines did not define what constituted consensus.

In conclusion, there are major weaknesses in the procedures used to determine consensus-based recommendations for all three guidelines. These are lack of independence in decision making by experts, a lack of a formal mechanism for aggregating judgments, and a lack of diversity of expertise, particular in areas where consumers and carers could contribute and where cultural expertise is relevant.

While NHMRC gives quite detailed guidance on how to develop evidence-based recommendations, there is little guidance on best practice for developing consensus-based recommendations. While many of these weaknesses would be overcome by using formal consensus methods such as the Delphi process, there is a need for NHMRC and similar agencies to produce more rigorous quality standards for development of consensus-based recommendations.

On 2014 Oct 31, Wayne Butler commented:

The ABS does not recommend that the CTV be defined as prostate only. The consensus guidelines specify a target volume margin of 5 mm in all directions about the prostate except posteriorly. Refer to the second paragraph on page 10 of the paper by BJ Davis et al., "American Brachytherapy Society consensus guidelines for transrectal ultrasound-guided permanent prostate brachytherapy." Brachytherapy, 11:6-19, 2012.

On 2016 Sep 19, Saurabh Gupta commented:

More than 100 cases and variety of clinical presentation.

But developmental biologists & embryologists are having tough time to match these numbers.

Where are all fifth arteries in the human or mouse embryos?

On 2015 Jan 12, Larry Parnell commented:

This paper was my selection for Paper of the Week for 3-7 Nov 2014.

On 2014 Nov 17, Mick Watson commented:

Hi Robert, and thanks for the response. The results from your qPCR are inconclusive, really the only way to rule out contamination is to sequence the negative control, which has ideally been prepared using the same batch of kits and reagents. Also, the fact that the virus has an effect in mice is negated if the virus is a contaminant, as it may never get into mice in a natural environment.

May I ask why you didn't sequence the negative controls?

On 2014 Nov 06, Robert H Yolken commented:

We actively considered contamination as the source of the sequences we obtained since these viruses may be common in the environment. However, we believe that contamination is rendered unlikely by the fact that, in many cases, we were able to document the presence of DNA homologous to ATCV-1 by 2 independent methods, library generation and quantitative PCR. In the quantitative PCR the reagent controls gave consistently negative results. We also believe that the plausibility of our findings in humans is supported by the mouse experiments presented in the publication.

On 2014 Nov 04, Mick Watson commented:

I'd like to politely suggest that the authors need to rule out contamination from reagents and kits used in the experiments (e.g. see http://biorxiv.org/content/early/2014/07/16/007187)

On 2015 Feb 09, Peter Csermely commented:

Authors of the paper are happy to highlight the available additional information regarding the localization tree, manual mapping and revision, as well as the updates of the ComPPI database http://comppi.linkgroup.hu.

Localization tree: ComPPI uses the GeneOntology (GO) cellular component terms for standardizing the different subcellular localization nomenclature in the source databases. The localization tree was needed for the unambiguous classification of minor localizations to the major subcellular compartments. Mappings of the original subcellular localization names to the relevant GO cellular component term for all input sources are available on our GitHub repository https://github.com/erenon/ComPPI/wiki/LOC-databases, while the whole localization tree and the mapping of minor localizations to major cellular components is accessible via our respective Help page http://comppi.linkgroup.hu/help/subcell_locs#structure, and was included in the Supplement of the paper http://nar.oxfordjournals.org/content/43/D1/D485/suppl/DC1 as Figure S2 and Table S4.

Manual mapping of proteins: As we described it in our published paper (Figure 1) we manually built a mapping table to map 30% of proteins in ComPPI when 1.) the original gene ID (e.g. HGNC gene symbol, EnsemblGeneId etc.) was translated to more than one UniProt accessions (~20% of proteins total, e.g. "HTT" gene symbol could be mapped to both P31645 and P42858 UniProt accessions, where our manual decision of P42858 was based on the name description, "Huntingtin" found in the source dataset of OrganelleDB); or 2.) the original gene ID had no automatic translation to UniProt ID (~10% of proteins total, e.g. manual mapping of the OrganelleDB source dataset "p53AIP1" gene name to Q9HCN2 UniProt accession having the official gene ID "TP53AIP1"). We note that the process was not entirely manual, as we used online ID cross-reference tools (such as PICR http://www.ebi.ac.uk/Tools/picr/) and the UniProt ID mapping http://www.uniprot.org/uploadlists/ as gold standard validation of gene ID to UniProt mappings.

Manual revision: Our manual revision process besides the construction of the localization tree and the ID mapping included 1.) the manual testing of the interfaces connecting the source databases to the ComPPI dataset during the database building process including the check for false-entries (e.g. non-existing UniProt accessions) and data content errors (e.g. minor subcellular localizations without GO terms) for randomly chosen 200 proteins, and 2.) six experts tested the integrated dataset for false-entries, data inconsistency and protein name mapping errors. Additionally, two of the six experts tested randomly chosen 200 proteins for exact matches between the entries in the source databases compared to the ComPPI dataset (this manual revision process is described in Figure 1 of the paper and in Supplementary Table S3). In Supplementary Table S3 of the publication we showed how many proteins, subcellular localizations and interactions were included from the source databases to the ComPPI dataset. The amount of data 'missing' in ComPPI compared to the sources is due to mapping differences between different namespaces (e.g. the integrated CCSB dataset contained 3,881 interactions, while only 3,733 interactions were included to the ComPPI dataset due to the removal of non-protein coding genes), data inconsistencies, or protein name mapping errors.

Database updates: The manual steps, such as the inclusion of new GO terms to the localization tree, or mapping of missing IDs, can and will be included to all future ComPPI updates. In these updates we will document our manual curation process in more detail. We schedule the release of ComPPI 2.0 before May 2016. The new version will include the update of the source databases (such as the 2014 update of the CCSB dataset), the revision of our mapping system with the update of the UniProt dataset based on the latest release, the inclusion of new input sources, such as LocDB, as well as an upgraded network visualization and extended export options (such as SIF or PSI-MI).

On 2015 Jan 27, Robert M Flight commented:

Commended because the issues they raise are valid. It makes sense that protein-protein interactions (PPI's) should be localization specific, and that many false PPI may be removed by considering localization information.

However, the solution implemented by ComPPI is not an ideal solution. This is largely due to the "manual" curation steps required in its construction. These manual steps are described below, followed by a comment on why this is such a problem for this type of resource.

Localization tree: the authors' state that 1600 of the ~3600 gene ontology (GO) cellular compartment (CC) terms were used to "manually" construct a localization tree whereby each GO term has a single path to one of the six defined compartments of Cytosol, Nucleus, Mitochondrion, Secretory-Pathway, Membrane, and Extracellular. There is no justification for the requirement of a tree with single paths to root compartments beyond the multi-parent nature of the directed-acyclic-graph of the normal GO. The description in the paper implies that the generation of the tree from the GO terms was done fully by hand. No description of how terms are assigned to compartments are provided, or if any attempts at automated assignment methods were made.

Manual mapping of proteins: The authors state that 30% of protein IDs were un-mappable to UniProt IDs using databases, and had to be mapped manually. Although protein-protein ID mapping across databases is known to be a difficult problem (I will point to one possible solution: http://bioinformatics.louisville.edu/abid/), this seems like a rather large percentage to map by hand.

Manual revision of database: 200 entries were checked by six experts, and based on their findings, revisions of the database made. No description of the type of checks made is provided, nor numbers on the rates for true, false / positives / negatives, and what the corrective efforts involved.

Each of these manual steps is a possible point of introducing bias that is hard to quantify and replicate, and in fact makes it highly unlikely that the database will be updated at regular intervals in the future or expanded to other model organisms beyond yeast, c elegans, drosophila and human. This severely limits the reliability and future utility of the database in my opinion.

On 2014 Nov 01, Abduzhappar Gaipov commented:

well

On 2015 Aug 12, Jim Woodgett commented:

This manuscript would be improved by systematic removal of "beta" throughout as the inhibitor primarily used, GIN, is not selective for GSK-3beta vs the related isoform, GSK-3alpha. A direct quote from reference 17 (referring to the source of the GIN inhibitor used here):

"A similar assay measuring inhibition of GSK3-alpha was also routinely run, but 7-12 showed no significant ability to discriminate between the two isoforms of GSK3 (data not shown). To the best of our knowledge, no isoform specific inhibitors of GSK3 have been reported, probably due to their high sequence homology (vida supra)."

This inhibitor (GIN) acts equally on both isoforms of GSK-3 (alpha and beta) and should not be referred to as a GSK-3beta selective inhibitor. This makes sense given that inhibition of GSK-3beta alone is insufficient for deregulation of beta-catenin (PMID: 17543867).

On 2014 Dec 02, Mikkel Wallentin commented:

On the behavioral effects, the authors write: "Compared with the low-flavanol intervention, subjects who received the high-flavanol intervention showed a mean improved cognitive performance of 630 ms." However, visual inspection of the supplementary figure 3 clearly shows that the 630 ms difference at follow up is due to a modest improvement of approx. 200 ms between baseline and follow up and a much more pronounced DECLINE in the control group (approx. 400ms). Needless to say, such a difference cannot be interpreted as "a mean improved cognitive performance of 630 ms", if it can be interpreted at all.

On 2014 Oct 27, David Colquhoun commented:

For all the reasons given by Hilda Bastian (and a few more, like P = 0.04 provides lousy evidence) it astonishes me that this study should have been trumpeted as though it represented a great advance. That's the responsibility of Nature Neuroscience (and, ultimately, of the authors).

I wonder whether what happens is as follows. Authors do big fMRI study. Glamour journal refuses to publish without functional information. Authors tag on a small human study. Paper gets published. Hyped up press releases issued that refer mostly to the add on. Journal and authors are happy. But science is not advanced.

I certainly got this impression in another recent fMRI paper in Science. Brain stimulation was claimed to improve memory (P = 0.043)

I guess these examples are quite encouraging for those who think that expensive glamour journals have had their day. Open access and open comments are the way forward.

On 2014 Oct 27, Hilda Bastian commented:

This report of a very small, short-term trial in healthy adults does not meet the CONSORT standards for trial reporting in several key respects. It does not provide sufficient data on the cognitive outcomes assessed, nor an adequate flow chart of outcomes (despite considerable attrition). There is also very little detail provided in the record of this trial at ClinicalTrials.gov.

The abstract does not make it clear that this is a dietary supplement and exercise trial (partially funded by a manufacturer). There were apparently two cognitive outcome measures on a ModBent task (an adapted test not elsewhere validated): immediate matching and delayed retention. Both relate to very specific functions, not an overall rating of cognitive abilities.

No effect was found for the exercise component in the trial, and out of the two cognitive measures, some effect was found for one, but not the other. That this is a chance finding surely can't be ruled out.

This report describes low vs high supplement groups. The study in ClinicalTrials.gov for the trial number they provide, however, was for a supplement and a placebo comparator.

Despite the major limitations of this single trial to address the question, the "Newsroom" report for the trial claims that it shows that "dietary flavanols reverse age-related memory decline."

It's good to see claims about dietary supplements tested. However, the results here rely on a chain of yet-to-be-validated assumptions that are still weakly supported at each point. In my opinion, the immodest title of this paper is not supported by its contents.

On 2015 Jul 12, Guo-Liang Jiang commented:

Evaluation of the value of ENI in radiotherapy for cervical and upper thoracic esophageal cancer: a retrospective analysis (Radiat Oncol. 2014 Oct 25;9:232.) We correct the first author Liu M's affiliation as the following: 1. Department of Radiation Oncology, Fudan University Shanghai Cancer Center, Shanghai 200032, China; 2.Department of Oncology, Shanghai Medical College, Fudan University, Shanghai 200032, China; Current address: Department of Radiation Oncology, Shanghai Chest Hospital, Shanghai Jiao Tong University, Shanghai 200030, China

On 2017 May 02, Zvi Herzig commented:

The authors note, "Observed trends require confirmation in a larger nationally representative sample". This has become possible with Poland's Social Diagnosis data (http://diagnoza.com/index-en.html).

The national data indicates that sharp declines in youth smoking have occurred during the present study's duration.

Below are the are smoking rates for ages 16-24, as reported in the 2013 (p. 249) and the 2015 (p. 284) Social Diagnosis reports:

year: smoking (%)

2000: 22.7

2003: 23.3

2005: 21.4

2007: 22.7

2009: 21.6

2011: 19.4

2013: 18.1

2015: 15.5

The differences between this local study and the national trend may reflect the fact that only 13 of the 25 schools surveyed have participated in both the 2010-11 and the 2013-14 surveys.

On 2015 Mar 16, David Ayoub commented:

That Contreras et al1 failed to report an association between vitamin D deficiency (VDD) and fracture risk is surprising in light of nearly a century of research linking vitamin D to bone health. We would like to suggest that study design limitations might have obscured such a potential relationship.

While fractures are a well-known complication of rickets/osteomalacia their association with subclinical forms of deficiency is less well established. The prevalence of fractures in rickets varies but typically found in a minority of individuals suffering from the disease. In a review of vitamin D and skeletal health in children, Moon et al recently summarized 17 modern studies of rickets. Fractures were observed in 13 of these studies among 1,177 children. The average fracture prevalence was 6% with a range between 0% and 20%. Only 2 studies involved a similar aged subset as reported by Contreras. Agarwal and Gulati reported a 6% fracture prevalence in 10-13 y/o children in India. Narchi et al reported no fractures among 10-15 year olds with rickets in Saudi Arabia. A comparison of fracture rates among vitamin D sufficient vs. deficient individuals would therefore require an adequately large sample size ideally in prospective studies. Contreras’ alternative approach that compared VDD rates among those with fractures and non-injured cohorts introduces several potential confounders.

Bone failure, especially with minor trauma, is considered to be a stochastic event – one associated with probabilities – caused by the interplay of various extrinsic (force/load) and intrinsic (bone strength) factors. Intrinsic factors put genetically susceptible individuals at risk for fractures when exposed to the random forces of trauma. The genetic component has been reported to be the most important intrinsic factor associated with bone failure. Other risk factors include, but are not limited to decreased bone loading, illness, various endocrine and nutritional conditions, medications, physiologic alterations associated with bone turnover, and growth spurts. Designing a population-based study to isolate the role of VDD in fractures would be nearly impossible unless other major determinants that effect bone quality and fracture risk are controlled.

Contreras provides little detail of the nature of forces applied to the 100 fracture cases. Even if vitamin D deficiency was the major contributor to diminished bone strength, a force is still required to produce a fracture. Contreras’ fracture group could have had other confounding conditions that affected bone strength, including familial predispositions that adversely influenced bone quality. Since forces vary considerably it is certain that some fractures would occur regardless of bone strength. The fact that 63% of cases with fractures following “minor” trauma had inadequate vitamin D levels would raise concern that diminished bone strength may have contributed to injury in some children.

It is possible that Contreras’ control population, similarly suffering from VDD ubiquitous to the general population, also had compromised bone strength and differed only from controls by having the good fortune of avoiding a significant accident. It would have been more ideal to choose a control group strictly comprised of children suffering traumatic injuries but without fractures instead of a group that included various acute medical conditions. In this scenario, all subjects of both groups would have been exposed to physical forces that would have challenged bone integrity. It is possible that their control group of emergency room patients without fractures suffered from conditions associated with higher rates of VDD and thus neutralizing any potential difference in vitamin D status from the fracture group.

In addition to Ryan, there are several more studies that did report an association between vitamin D and fractures. Ruohola et al reported a strong association between lower vitamin D levels and stress fractures among 800 randomly selected and prospectively followed young military recruits. Leboff et al reported lower mean vitamin D levels in women with postmenopausal hip fractures compared to two groups of postmenopausal women prior to elective hip replacements. In a nested case-control study of 1200 female Naval recruits Burgi et al reported increased risk of stress fractures among those who were vitamin D deficient. There was approximately half the risk of stress fractures in women in the top vs. the bottom quintile of vitamin D concentration. While we agree that one cannot say that vitamin D status alone could predict who will fracture, these alternatively designed studies suggest that it can identify an at-risk population.

Lastly, but perhaps of greater importance is the extraordinary rate (63%) of inadequate vitamin D status among Contreras’ emergency room-attended population. This observation alone deserves the full attention of local physicians and public health officials and goes far beyond the original mission of their study.

REFERENCES 1. Contreras JJ, Hiestand B, O'Neill JC, Schwartz R, Nadkarni M. Vitamin D deficiency in children with fractures. Pediatr Emerg Care. 2014;30(11):777-781. 2. Moon RJ, Harvey NC, Davies JH, Cooper C. Vitamin D and skeletal health in infancy and childhood. Osteoporos Int. 2014;25(12):2673-2684. 3. Agarwal A, Gulati D. Early adolescent nutritional rickets. J Orthop Surg (Hong Kong). 2009;17(3):340-345. 4. Narchi H, El Jamil M, Kulaylat N. Symptomatic rickets in adolescence. Arch Dis Child. 2001;84(6):501-503. 5. Ferrari S, Chevalley T, Bonjour JP, Rizzoli R. Genetic determinants of bone microstructure and fracture risk in childhood. Bone. 2007;40(3)(suppl 1):S12. 6. Melamed ML, Kumar J. Low levels of 25-hydroxyvitamin D in the pediatric populations: prevalence and clinical outcomes. Ped Health. 2010;4(1):89-97. 7. Ryan LM. Forearm fractures in children and bone health. Curr Opin Endocrinol Diabetes Obes. 2010;17(6):530-534. 8. Ruohola JP, Laaksi I, Ylikomi T, et al. Association between serum 25(OH)D concentrations and bone stress fractures in Finnish young men. J Bone Miner Res. 2006;21(9):1483-1488. 9. LeBoff MS, Kohlmeier L, Hurwitz S, Franklin J, Wright J, Glowacki J. Occult vitamin D deficiency in postmenopausal US women with acute hip fracture. JAMA. 1999;281(16):1505-1511. 10. Burgi AA, Gorham ED, Garland CF, et al. High serum 25-hydroxyvitamin D is associated with a low incidence of stress fractures. J Bone Miner Res. 2011;26(10):2371-2377.

On 2014 Dec 30, William Grant commented:

The review by Bonifazi and colleagues found that those with sarcoidosis have an increased risk of many types of cancer.1 The role of inflammation in explaining the link between sarcoidosis and cancer was discussed. However, no firm conclusion was reached regarding the link.

A factor not considered was vitamin D. Those with sarcoidosis often have low 25-hydroxyvitamin D [25(OH)D] concentrations due to concerns about hypercalcemia.2 There is evidence that low solar UVB doses and serum 25(OH)D concentrations are a risk factor for sarcoidosis. A summary of the geographical and temporal mortality rates for sarcoidosis in the United States for the period 2000-2007 found higher mortality rates for non-Hispanic black and white decedents in the higher latitude states, which have lower solar UVB doses, as well as moderately higher trends of mortality rates for non-Hispanic males and females from 1989 to 2007.3 Serum 25(OH)D concentrations decreased during that period due to a number of factors.

Low solar UVB exposure and 25(OH)D concentrations are an important risk factor for many types of cancer.4 Of the types of cancer with significant relative risks related to sarcoidosis, seven are significantly inversely related to solar UVB doses in ecological studies: colorectal, kidney, leukemia, liver, and upper digestive tract cancer, Hodgkin's lymphoma, and non-Hodgkin's lymphoma. However, eight types of cancer inversely correlated with solar UVB doses in ecological studies were not found directly linked to sarcoidosis: bladder, breast, cervix/uterus, lung, ovary, pancreas, prostate, and stomach cancer. Of these types, only bladder and breast cancer are strongly linked to low UVB doses and/or 25(OH)D concentrations. As noted in Ref. 1, smoking may affect the sarcoidosis-cancer link, and four of these types of cancer are strongly linked to smoking: bladder, crevix/uterus, lung, and pancreas cancer.

Twelve of the 16 studies included in Ref. 1 were from Nordic countries. A study of cancer incidence rates with respect to a solar UVB exposure index in Nordic countries based on 54 occupational categories found much the same for males as summarized in Ref. 4.5 In addition, the melanoma incidence rate for males was inversely correlated with the UVB exposure index at the p=0.02 level, and the non-melanoma skin cancer rate was significantly inversely correlated with both the UVB index and lung cancer incidence rates in a multiple-linear regression analysis. Both melanoma and basal cell carcinoma are more frequent among people with limited solar UV exposure than those with chronic UV exposure.

References 1. Bonifazi M, Bravi F, Gasparini S, et al. Sarcoidosis and cancer risk: systematic review and meta-analysis of observational studies. Chest. 2014 Oct 23. doi: 10.1378/chest.14-1475. [Epub ahead of print] 2. Bolland MJ, Wilsher ML, Grey A, et al. Randomised controlled trial of vitamin D supplementation in sarcoidosis. BMJ Open. 2013;3:e003562. 3. Swigris JJ, Olson AL, Huie TJ, et al. Sarcoidosis-related mortality in the United States from 1988 to 2007. Am J Respir Crit Care Med. 2011;183:1524-1530. 4. Moukayed M, Grant WB. Molecular link between vitamin D and cancer prevention. Nutrients. 2013;5:3993-4023 5. Grant WB. Role of solar UV irradiance and smoking in cancer as inferred from cancer incidence rates by occupation in Nordic countries. Dermatoendocrinol. 2012;4:203-211.

Disclosure I receive funding from Bio-Tech Pharmacal (Fayetteville, AR), the Sunlight Research Forum (Veldhoven), and Medi-Sun Engineering, LLC (Highland Park, IL).

On 2014 Oct 24, Fernando Castro-Chavez commented:

To All,

When I was comparing the relative position of the 64 codons of the Genetic Code between two x-y 2-D Tables, i.e., while keeping the axis Y constant with the C-Rings of the DNA Nucleotides and changing the axis X from having their H-bonds in one Table to their Tautomerism in the other Table, what resulted of this comparison, as my article demonstrates, were the directional arrows of the ancient Chinese representation of the Yin and the Yang (external long arrows at opposite directions annealed to the two internal shorter arrows per each of the long ones). When I added the third Table with the third possible comparison of these DNA-Nucleotide properties (i.e., H-bonds in axis X and Tautomerism in axis Y), what I had was the half of a Cube, and by adding the remaining three reciprocal Tables, I was able to complete a Cube. From here, it was not difficult to Spherize such Cube in order to obtain the Spherical representation of the genetic code!

This is my graphic representation of one of the Spherized sides of the Cube: Spherical Genetic Code

Attentively,

Fernando Castro-Chavez. 10/24/2014 Houston, TX

On 2014 Oct 27, Jonathan Eisen commented:

A version of this paper is available, freely and openly, in arXiv: http://arxiv.org/abs/1401.5130

On 2014 Nov 30, Harri Hemila commented:

Musher DM, 2014 conclude their paper by stating that RCTs are needed to determine whether the antiinflammatory activity of macrolides or statins is beneficial in treating CAP.

I would like to propose that RCTs are also needed to determine whether vitamin C is beneficial in treating CAP. In dozens of animal studies, vitamin C protected against infections by various viruses and bacteria, implying that the vitamin may influence incidence and severity of some infections Hemilä 2006, pp. 5-9, 105-21.

In the early 20th century, Alfred Hess carried out extensive studies of scurvy and summarized a large series of autopsy findings as follows: “pneumonia, lobular or lobar, is one of the most frequent complications [of scurvy] and causes of death” and “secondary pneumonias, usually broncho-pneumonic in type, are of common occurrence, and in many [scurvy] epidemics constitute the prevailing cause of death”, see Hemilä H, 2007. In this journal, Hess 1932 commented that in “infantile scurvy … a lack of the antiscorbutic factor which leads to scurvy, at the same time predisposes to infections [particularly of the respiratory tract]. … Similar susceptibility to infections goes hand in hand with adult scurvy.” Thus, deficiency of vitamin C and pneumonia may be associated. Deficiency of vitamin C is not rare even nowadays Schleicher RL, 2009.

Furthermore, vitamin C has reduced the duration and severity of the common cold in well-nourished people suggesting that the vitamin may have effects on the respiratory system even in the absence of frank deficiency Hemilä H, 2013.

Hemilä H, 2013 carried out a Cochrane review and found 3 controlled trials that looked at whether vitamin C prevents pneumonia and 2 that looked at whether it might help in curing pneumonia. Each of the 5 studies found that vitamin C supplementation was significantly beneficial against pneumonia. Two of them were placebo-controlled RCTs.

Pitt HA, 1979 administered vitamin C to US marine recruits and reported 7 cases of pneumonia in the placebo-group compared with 1 case in the vitamin C group. Hunt C, 1994 administered vitamin C to elderly people who were admitted to hospital in the UK because of bronchopneumonia or acute exacerbation of chronic bronchitis. They reported a significant decrease in the “total respiratory score” by vitamin C administration, and 5 deaths in the placebo group compared with 1 death in the vitamin C group. Hunt et al. tested the effect of vitamin C “over and above those of normal medication (mainly antibiotics and cough medicines) to which all participants were exposed” so that all their patients received antibiotics and vitamin C was not an alternative to them.

The prophylactic use of vitamin C to prevent pneumonia should be further investigated in populations who have a high incidence of pneumonia, especially if dietary vitamin C intake is low. Similarly, the therapeutic effects of vitamin C should be studied, especially in pneumonia patients with low plasma vitamin C levels.

On 2014 Oct 24, Gwinyai Masukume commented:

The authors write, "Only two cases of an abdominal pregnancy combined with an intrauterine pregnancy has been reported [11, 12]."

Zacchè MM, 2011 is an additional recent case that the authors did not cite. In addition Reece EA, 1983 is a review of 589 cases of combined intrauterine and extrauterine gestations. A Chinese article Wu X, 1999 also tackles similar cases.

On 2015 Jul 15, Raphael Levy commented:

Thanks Harald for commenting on my blog post about SmartFlares / Nanoflares with reference to this paper.

I reproduce the conversation below. I hope it continues and others join in.

Raphael

"Hi everybody, as the correspending author of a Stem Cell paper in which we have used the SmartFlares on different pluripotent cells of human, murine and porcine origin I want to reply to two of the above mentioned questions.

Why do we see a signal at all in the scramble control? I think one cannot expect a negative control which does not produce a fluorescent signal at all. The fluorophore may not be quenched by 100% and may be subject to degradation, especially when applied for a longer time (two days or more). Nevertheless, within 16 to 24 hours after the application of the nanoparticles we see a clear-cut difference of fluorescence intensity when comparing scramble control and gene-specifc Smart Flares. http://www.ncbi.nlm.nih.gov/pubmed/25335772

We believe that this difference is reliable and specific. We have selected freshly reprogrammed murine iPS cells based on their Nanog-specific fluorescence intensity in situ. In downstream experiments we could confirm that only colonies with a high fluorescence intensity expressed higher amounts of endogenous pluripotency factors and showed a superior capacity to differentiate. Therefore, we belive that these functional data strongly support the idea that the fluorescence intensity was indeed correlated to a specific interaction with the Nanog mRNA in these clones.

Why do different cells take up varying amounts of SmartFlares? I think this difference is not surprising as the nanoparticles are engulfed by endocytosis. This process is influenced by the cell type, the differentiation status and the cellular ability to perform phago- and macropinocytosis. Therefore, we think that a uniform uptake rate cannot be expected."

I replied "Hi Harald

Thanks again for commenting here and sorry for the delay in replying. It is interesting that you see some differences but the big question that remains is how could the technology possibly work?

It can only work if the particles escape endosomes, but: 1) there is no reason why they should, 2) this problem is not discussed in the original publication introducing the technology, 3) there is no direct evidence in the literature that it happens, and, 4) all the data we are accumulating indicates that the particles are indeed in vesicular compartments (more on this soon on the open notebook as we have just had our cell electron microscopy results this week).

The images shown in your articles are low mag overviews of many cells and therefore the resolution does not allow to discuss any cellular localization. Do you have any higher resolution images that you could share? Do you have any (direct) evidence and/or proposed mechanism for endosomal escape?

The unequal distribution of uptake (cell to cell variability) is also a big concern. I don鴠believe that it relates to differences between rate of uptake of different cells. Such differences would average over an 18 hour period and they should also be seen in the dextran uptake. A possible interpretation would be some degree of nanoparticle association/aggregation before interaction with the cells (this is to be tested experimentally).

Raphael"

On 2017 Jun 22, Nathaniel Huebsch commented:

The original url for downloading the software is no longer active. The correct website for downloading this software is:

https://gladstone.org/46749d811

We appreciate everyone who let us know about the weblink being down.

On 2014 Dec 14, Jeffrey Beall commented:

I would like to express some concerns regarding this article. Examining the publisher's PDF version of the article, one finds this statement after the abstract: "Submitted Apr 19, 2014. Accepted for publication Apr 22, 2014." Based on this three-day period between submission and acceptance, it is fair to conclude, I think, that no bona fide peer review was carried out.

Moreover, it appears that the article is being posted to online patient forums, perhaps by one of the authors, as a way of advertising and marketing the surgical technique described in the article.

On 2015 May 04, Andrew Brown commented:

See our letter about this article here Brown AW, 2015; and the authors' response here http://jn.nutrition.org/content/145/5/1029 (not yet indexed in PubMed).

On 2016 Apr 27, Nadav Sorek commented:

After the paper was published, we became aware of anomalies in the ITC plot in figure 2, and in the gel blot presented in figure 3. I performed the experiment and prepared the figure, and was not aware of these anomalies, as they can be seen only when the contrast of the blot has been changed dramatically. The anomalies in the ITC plot were determined to have been caused by a failure in the graphics card. The reason for the anomalies in the gel blot remains unknown, although they may have occurred as a result of image compression into JPEG format. I did not retain the original uncompressed images, so this was impossible for me to resolve without repeating the experiment. I was able to demonstrate that the results reported were reproducible, however due to the anomalies, JBC insisted that we retract the paper. To remove any doubt about the reliability of the data, we asked an independent researcher to repeat the experiment, and the results from these experiments were consistent with our published results. The report with all the details can be found in bioRxiv (http://biorxiv.org/content/early/2016/04/26/050427). Please don’t hesitate to contact me with question or further clarifications.<br> Nadav Sorek (Nadav.sore.il@gmail.com)

On 2015 Jan 12, Larry Parnell commented:

This paper was my selection for an alternate Paper of the Week for 3-7 Nov 2014.

On 2016 Dec 10, Wen-Long Hu commented:

Laser acupuncture (LA) is defined as the stimulation of traditional acupuncture points with low-intensity, non-thermal laser irradiation. LA combines the positive effects of traditional acupuncture with LLLT. http://dx.doi.org/10.5772/55092

On 2016 Dec 01, Jan Tunér commented:

In the study by Hu et al., three acu points and 2-4 ashi points were irradiated, all but one (Hegu, LI4) were in the area of the masticatory muscles. So in fact, laser acupuncture and traditional LLLT for TMD was used. Let us look at the laser parameters: The laser was a 810 nm 150 mW laser, “chopped” at 50%, so actually 75 mW. Reason for chopping is not presented. Two acu points were in the TMD area: ST7 = condyle, ST6 = mandibular corner = muscle masseter. 5 s per acu point = 0.375 J. Two points in the TMD area = 0.750 J. 40 s per ashi/tender point = 3 J. For two points 6 J, for four points 12 J. Maximum total 12.750 J. Several studies on TMD and LLLT have used energy ranges between 3 and 6 J per point, or even less, and targeting tender points. Therefore, this study is a traditional LLLT study and the contribution of the acu points cannot be evaluated. Future studies should not mix local LLLT with laser acupuncture.

On 2015 Apr 02, Alasdair MacLullich commented:

Thank you for the clarification. This is a very important issue because low arousal states of acute onset (excluding coma) are considered as indicating "severe inattention" in DSM-5: see the DSM-5 guidance notes.

Many patients with low arousal states (of acute onset) are not testable by conventional cognitive tests, and yet these patients mostly have delirium. Because of this, there is an explicit 'untestable' category in the 4AT, a rapid assessment test for delirium designed for use in routine clinical practice (see www.the4AT.com; http://www.ncbi.nlm.nih.gov/pubmed/24590568; http://www.ncbi.nlm.nih.gov/pubmed/23988641).

The issue of low arousal states and delirium diagnosis was covered in a consensus statement by the European Delirium Association and the American Delirium Society: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4177077/

See also these two relevant papers: http://www.ncbi.nlm.nih.gov/pubmed/24080383; http://www.ncbi.nlm.nih.gov/pubmed/22173963

On 2015 Mar 15, Geriatric Medicine Journal Club commented:

Thank you so much. We will share this comment with a link to the followers of #GeriMedJC on Twitter.

On 2015 Mar 09, Edward R Marcantonio commented:

A recent Geriatric Medicine Journal Club commented on the limitations of the 3D-CAM in low arousal states. Note that for each 3D-CAM question, "no response" is considered the equivalent of "incorrect" and can trigger the presence of a CAM Feature. Therefore, the 3D-CAM should work equally well, and be even quicker, in patients with low arousal states. While we had relatively few patients with low arousal states in our validation study, the 3D-CAM correctly identified 100% of the delirium cases in this subgroup.

On 2015 Feb 27, Geriatric Medicine Journal Club commented:

This study of the 3D-CAM, a 3-minute diagnostic assessment for CAM-defined delirium, was critically appraised at the November 2014 Geriatric Medicine Journal Club (follow #GeriMedJC on Twitter). Much discussion was generated between researchers and clinicians from around the world. See the full transcript of the discussion here: http://gerimedjc.blogspot.com/2014/11/gerimedjc-november-28-2014.html?spref=tw Highlights included limitations of assessment in patients with low-arousal states.

On 2015 May 31, Marc Girard commented:

In accordance with my previous criticism regarding the methodological reliability of most studies presented as confirming the safety of vaccines, this investigation raises a number of serious concerns.

Case ascertainment – Whereas the study title makes special emphasis on multiple sclerosis (MS: ICD code 340), case identification includes no less than nine ICD codes, some of which (optic neuritis or acute disseminated encephalomyelitis being sometimes difficult to differentiate from genuine MS, whereas others [transverse myelitis] are generally considered as distinct). The most expected result of such a diagnosis blending is to weaken statistical power and to blur epidemiological evidence.

Vaccination assessment – Only 4.0% of the 3885 controls were exposed to hepatitis B vaccine in the 3 years before the index rate; this may be compared with the study by Hernán MA, 2004 (the design of which was fairly similar), where 2,4% of the 1565 controls were exposed to a recombinant hepatitis B vaccine. The trouble is that this immunization was highly selective in the latter population (UK), whereas it was massive in the former (USA). In spite of this major discrepancy in the vaccine policy between the two countries, the surprisingly small difference between these two percentages raises the hypothesis that, for one reason or another, vaccination recording was incomplete in the American sample. Although duly pointed out as remarkable by Langer-Gould et al., low vaccine exposure in their sample was not seriously discussed by the authors.

Control selection – Although a black ethnicity was the most prominent risk factor identified by the authors in their previous study on the incidence of demyelinating syndromes (quoted as reference 17 in their current paper), one may wonder why their control selection did not include race in their matching method. As it happens, imbalance in the distribution of black race between cases and controls was the most striking feature of the baseline samples characteristics.

Index date – Although the timing of symptoms appearance is generally a crucial argument for causality in drug monitoring (there may be exceptions to this rule), this parameter is never properly considered in investigations devoted to post-vaccine MS. Actually, as the disease may remain clinically silent for years, the relevant parameter is neither the date of diagnosis nor that of the late symptoms which lead to the investigations leading to positive diagnosis. In spite of this, what investigators mean by “symptoms onset date” is never clearly defined: which symptoms? For example, in their abovementioned reference 17 (Table 1), Langer-Gould et al. estimated at 0.9 month the median time from symptom onset to diagnosis, after having stipulated that, defining MS required two or more episodes of MS “separated in time”: is unlikely that 0.9 month is a sufficient time interval to separate two distinct MS episodes… At the opposite side of the clinical spectrum, the very first symptoms of a MS are often an unexplained fatigue, mild paresthesia, etc. the onset of which may be quite close to the time of vaccine injection (a few days or weeks), but which may last for years before onset of more significant symptoms: thus, if one focus on the late significant symptoms, this very long time lag is almost always interpreted as speaking against a vaccine role whereas, when considering the whole of symptoms sequence from its very beginning (i.e. from the time of quite discrete symptoms just after injection), it is on the contrary highly suggestive of a vaccine causality. I have never seen this crucial problem properly taken into account in any database, so that most investigations about the time between vaccination and the onset of MS symptoms are essentially misleading.

Regarding MS and in spite of their denials, the authors ended up to a result very close to that of Hernan et al.’s., namely an overrepresentation of cases (4.2%) as compared to the controls (3.1%) within a time windows of 3 years. Of course, this difference just failed to reach statistical significance but: i) as documented above, the methodological tendency of the authors contributed to decrease the power of their results; ii) amongst the published case/control studies supposed to exclude a post-vaccine risk of MS (by means of like strategies of dilution of the cases or of insufficient observation period), the number of those suggesting (even in a nonsignificant way) an overrepresentation of cases in vaccinated subjects is clearly higher than those suggesting an underrepresentation, and the difference between the two groups of studies is clearly significant from a statistical point of view.

Finally and as with most papers devoted to the safety of hepatitis B vaccines, the authors cannot refrain from concluding that no “change in vaccine policy” is warranted: yet, their investigation is totally devoid of the slightest element likely to validate any vaccine policy, whose potential shortcomings (included issues of cost, of resources allocation, of individual and collective efficacy, of nonneurological risks, etc.) go far beyond the sole issue of MS. In psychoanalysis, such optimism (going far beyond the available evidence from a given investigation) is called “the return of the repressed”…

On 2015 May 28, Andrew Brown commented:

See our comment about the analysis of this cRCT in our letter to the editor published by the journal: "Ignoring the potential similarity among individuals in the same cluster (school) can ... increase the type I error rates and jeopardize the validity of conclusions from cRCTs." http://www.hindawi.com/journals/jobe/2015/708181/

On 2014 Oct 21, Carl L von Baeyer commented:

Potentially a very important finding. However, it should be read with caution. Table 2 indicates a range of 6-8 for VAS. This is problematic in four ways: (a) VAS is not listed among the measures, which were a faces scale and a numerical rating scale. (b) Perhaps what is meant by "VAS" is actually a combination of scores on the faces scale and the numerical rating scale. But these different scales cannot be combined as if they were the same thing. They should be reported separately unless a rationale is provided for treating them as the same. (c) The idea that ALL 82 subjects with a chronic illness used only the scores 6, 7 or 8 (out of the 11 scores available on the self-report scale) is not plausible: this would represent an extraordinarily unlikely restriction of range on the 0-10 metric, or else there was some bias in the way the question was asked. (d) If all scores were 6, 7 or 8, then the mean could not have been greater than 8 as reported in the abstract. I'd invite the authors to check this line of Table 2 before the paper is finalized. Thank you.

On 2015 Jan 06, George McNamara commented:

Check out Dr. Jahan Khalili's take on this research, at

https://www.immunodiet.com/evidence-for-peanut-avoidance-after-a-cancer-diagnosis/

On 2014 Oct 18, David Keller commented:

Restricted entry of travelers from affected regions is a valid means of pandemic control

Past pandemics have taught us that "travel restriction is an immediate and non-pharmaceutical means of retarding incidence growth. It extends the time frame of effective mitigation, especially when the characteristics of the emerging virus are unknown."(1)

During the 2014 ebola virus pandemic, the oft-repeated objection that travel restrictions would "exacerbate the West African epidemic by impeding the flow of aid workers and supplies" (2) could have been addressed by allowing aid workers to enter the ebola zones, and then return to the U.S. after appropriate observation or quarantine measures. Fortunately, the ebola pandemic did not break out into the U.S. population, despite transmission of the virus to hospital personnel.

Travel restrictions and quarantines are a valid means of slowing the spread of infection, and should be considered during the next deadly pandemic which lacks effective treatment.

References

1: Chong KC, Ying Zee BC. Modeling the impact of air, sea, and land travel restrictions supplemented by other interventions on the emergence of a new influenza pandemic virus. BMC Infect Dis. 2012 Nov 19;12:309. doi:10.1186/1471-2334-12-309. PubMed PMID: 23157818; PubMed Central PMCID: PMC3577649.

2: Gostin LO, Hodge JG Jr, Burris S. Is the United States Prepared for Ebola? JAMA. 2014 Oct 17. doi: 10.1001/jama.2014.15041. [Epub ahead of print] PubMed PMID: 25325877.

On 2015 Mar 24, Amanda Capes-Davis commented:

Thank you Radoslaw - I very much appreciate this clarification from you and the other authors of the article.

On 2015 Mar 23, Radosław Stachowiak commented:

Thank you for your vigillance and comments. To correct this important issue we have published Erratum to the article: http://link.springer.com/article/10.1007/s00284-015-0803-0?sa_campaign=email/event/articleAuthor/onlineFirst

On 2015 Feb 26, Amanda Capes-Davis commented:

Thank you to the authors for commenting on this paper in a feature article published in Science at http://www.sciencemag.org/content/347/6225/938.full. I appreciate the authors' comments in clarifying why these cell lines were used.

On 2015 Feb 13, Amanda Capes-Davis commented:

The three cell lines used here - INT-407, HEp-2 and HeLa - are all HeLa. INT-407 and HEp-2 were clearly shown to be cross-contaminated by HeLa in the 1960s and 1970s. INT-407 and HEp-2 continue to be used widely today, by scientists who may not be not aware that these cell lines are misidentified.

In my view, three cell lines are used here to indicate that the effect of Lmo0171 on cell morphology has broad impact across different cell types. But the results described here may be specific to HeLa cells and have no impact beyond that culture and cell type (human cervical adenocarcinoma).

I would urge authors to be more transparent regarding the cell line information that they provide to readers. The authors here list cell bank catalogue numbers in their Materials and Methods section. When you look at the catalogue entry for each cell line, the entries clearly state that INT-407 and HEp-2 are HeLa. The authors of this paper are therefore aware of that information.

In a letter to Nature back in 2000 (http://www.ncbi.nlm.nih.gov/pubmed/10667765), Stacey and other authors urged the research community to take action to improve our approach to misidentified cell lines. They recommend that all misidentified cell lines should be listed with the contaminant in brackets - for example, INT-407 (HeLa) and HEp-2 (HeLa). Taking that approach would make it much clearer to the reader that these three cell lines are all HeLa.

The International Cell Line Authentication Committee (ICLAC) curates a database of known misidentified cell lines. Readers are welcome to refer to the database as a resource when naming or using cell lines. The database can be found at http://iclac.org/databases/cross-contaminations/.

On 2014 Nov 12, Musharraf Jelani commented:

Before becoming Martin et al (2014) article online we had submitted an eletter to J Med Genet, however we did not receive any response for it publishing it online. We want to share our comments as follows.

"PLK4: A novel candidate for primordial dwarfism?"

Letter to Editor Shaheen et al. 1 recently reported a homozygous 5 bps deletion mutation (c.12991303delTAAG; p.Phe433Leufs*6) in the human polo-like kinase 4 (PLK4,MIM 605031) gene and proposed that it is a compelling candidate for primordial dwarfism (PD). Autozygosity mapping and LOD score, method of estimating linkage distances, are adopted for the novel locus identification. Candidate gene hunting among 144 in the region is based on filtering through ToppGene suite 2. Mutation screening through Sanger sequencing of PLK4 gene is only based on ToppGene suite ranking. In this study authors have not mentioned how many and which samples were selected for genotyping and linkage analysis so the predicted maximum LOD score cannot be calculated. Maximum multipoint LOD score (2.5) obtained by the authors is quite lower than the established threshold values (3.0). In article it has been stated that all PD genes in training set in comparison to all 144 genes of the locus in test set were enrolled in ToppGene candidate prioritization. These genes lists must be provided along with test parameters. In our opinion these informations are essential for data reproducibility and validation. Only PLK4 gene has been selected for Sanger sequencing but The candidate region (Chr4:112904466-129392060, GRCh37/hg19) includes some of the well- known syndromes characterized by autosomal recessive intellectual disabilities, microcephaly, short stature, and overlapping phenotypes. Alazami syndrome caused by mutations in LARP7, autosomal recessive type 1 mental retardation caused by mutations in PRSS12, Bardet-Biedl syndrome caused by mutations in BBS7 and BBS12, Van Maldergem syndrome 2 caused by mutations in FAT4, neuronal ceroid lipofuscinosis 7 caused by mutations in MFSD8, a multisystem disorder including brain function caused by mutation in SCLT1, as well as genes involved in mitosis and brain development e.g. MAD2L1, FGF2, INTU, have not been discussed throughout the manuscript. Furthermore, chromatograms for obligate carriers have not been presented nor the Saudi population screening for minor allele frequency has been performed. Similarly a homozygous variant (NM014264.4, c.1556G>C; p.Trp519Ser) reported in ESP6500 (http://evs.gs.washington.edu/EVS/) alters a highly conserved amino acid of PLK4 however it has not been assigned to any of primordial dwarfism phenotype and not discussed by the authors. In fact, phenotypic overlaps among the study mutants and the above syndromes would be of far more value instead of paying an exaggerated attention to PLK4 biology. References

1 Shaheen R, Al Tala S, Almoisheer A et al. Mutation in PLK4, encoding a master regulator of centriole formation, defines a novel locus for primordial dwarfism. Journal of medical genetics 2014. 2 Chen J, Bardes EE, Aronow BJ et al. ToppGene Suite for gene list enrichment analysis and candidate gene prioritization. Nucleic acids research 2009; 37: W305-11.

"conflict-of-interest" No conflict

On 2014 Oct 29, Benoit Kornmann commented:

This paper presents some spectacular features of membrane chemistry. If you are not into the topic, watching some of the featured movies will likely pique your interest.

It has long been known that different lipid species can have different affinity for each other, such that they may segregate into different phases, which can be visualized as subdomains on artificial membranes such as giant unilamellar vesicles (GUVs). Because a phase-separated membrane has a much lower entropy than a mixed one, phase separation can be achieved by lowering the temperature or increasing order in the membrane by stretching it.

In this paper, the authors bathed GUVs in a hypotonic solution. Because these GUVs are semi-permeable to water, they swell until they eventually rupture, causing a catastrophic leak that re-equilibrates pression. After the GUVs recover from rupture, they start swelling again, and go through several cycles of swelling-rupture until reaching osmotic equilibrium.

Here, the authors observe that, while in relaxed GUVs lipids are perfectly mixed, swelled GUVs show lipid phase separation. Mixing occurs right after each membrane rupture event; therefore, a cycle of separation-mixing follows the cycle of swelling-rupture of the GUV.

It was previously known that vesicles underwent cycles of swelling-rupture. It was also previously known that membrane tension favored lipid phase separation. It is also not clear whether these phenomena are relevant in the case of biological systems. Nonetheless, the movies presented here are spectacular and unveil unexpected and very complex dynamic behaviors of seemingly very simple chemical systems.

On 2015 Dec 01, Bastien Boussau commented:

My colleagues and I have published an article (Groussin M, 2016) in which we contest the results reported in the above study.

On 2014 Nov 22, Arturo Casadevall commented:

There is little that has not been said before in the most recent post by Joshua L. Cherry and thus we do not see the need for additional responses. Interested readers are referred to the original article and the comments above.

On 2014 Nov 20, Joshua L Cherry commented:

The reply above continues with the tactic that I criticized. It embraces and gives the benefit of the doubt to every argument in favor of the GOF experiments, ignores well-known counterarguments, and on this basis declares the other side's rhetoric to be misleading as though this were an additional pro-GOF argument rather than a corollary of the pro-GOF position. I will refrain from engaging in a rehash of arguments that should be familiar, but two specific points deserve brief mention. First, the reply glosses over the differences between laboratory engineered/selected transmission in ferrets and naturally-occurring human pandemic. Second, it makes a logical leap from somebody's intent to act on information to that information having value, as though nobody ever acted on valueless information.

On 2014 Nov 11, Arturo Casadevall commented:

We fear that our position has been mischaracterized in Joshua L. Cherry’s latest comment. For example, nowhere in our article or response do we say or imply that “GOF advocates are right and GOF opponents are wrong”. Although we believe that everyone is entitled to their own opinion, we also think that Joshua L. Cherry most recent comments merit an additional response.

First, we note that the correspondent is in fact invoking the ‘apocalyptic fallacy’ in his opposition to GOF experiments, something that we cautioned against in our editorial. The ‘apocalyptic fallacy’ arises when one side uses the apocalypse to inspire fear without factual data in the hope of convincing others of the correctness of their position. In our prior comment we chided both sides for using apocalyptic arguments but noted that that fallacy applied primarily to the anti-GOF camp because their appeals to fear were based on hypothetical accidents and scenarios, while the pro-GOF camp appeals to risk were based on known past epidemics. In his response Joshua L. Cherry mentions ‘reports of frequent laboratory accidents with infectious agents’ and ‘the return of H1N1 influenza from the laboratory in 1977’ to argue that anti-GOF proponents were in fact justified in their use of apocalyptic arguments. However, there is a problem here. First, while we are the first to agree that there is a need for utmost safety in these experiments, we note that it is guilt by association to aggregate GOF experiments with recent disparate laboratory accidents that range from a shipping error with H5N1 samples to incompletely sterilizing Bacillus anthracis spores. Second, the return of H1N1 1977 has been presumed by some to reflect the escape of that strain from a laboratory, but this has not been proven: that explanation for the reemergence of H1N1 remains a presumption that is not universally accepted among influenza experts. Thus, it is not reasonable to argue that these recent accidents should lead us to fear accidental pandemics from GOF experiments, whereas it is reasonable to worry that naturally occurring mutations in the H5N1 virus could lead to a global pandemic based upon historical evidence of similar such occurrences. Determining the possibility of precisely such events is the point of GOF experiments.

Second, Joshua L. Cherry also states that some ‘would question the likelihood of a natural H5N1 pandemic, and whether GOF experiments tell us much about this likelihood’. I think we can all agree that GOF experiments have shown that H5N1 has the potential to become mammalian transmissible and that that information is new. Since prior to those experiments there was debate among experts in the field as to whether this was even possible, then we must disagree with the statement that GOF do not tell us ‘much about this likelihood’. In fact, those experiments have shown unequivocally that the H5N1 has the biological potential to become mammalian transmissible and that fact alone implies a greater threat from a natural pandemic that we knew previously. At the recent meeting of the National Science Advisory Board for Biosecurity (http://osp.od.nih.gov/office-biotechnology-activities/biosecurity/nsabb), evidence was presented that information obtained from GOF experiments was in fact being used in strain surveillance and vaccine development. Hence, the argument from the pro-GOF camp that this information is in fact useful for preventing a pandemic is more than ‘purely theoretical’.

We agree with Joshua L. Cherry in his concerns about safety. In an earlier essay we took both sides to task for framing the debate in simplistic terms (PMID: 25085113) and we urge the need for careful analysis, rigorous thought and the avoidance of extrapolation and interpretation in framing arguments for and against this type of experimental work during the ongoing debate.

On 2014 Nov 04, Joshua L Cherry commented:

The response above seeks to justify criticizing "apocalyptic rhetoric" from one side of the debate while ignoring the same sort of rhetoric from the other side. The justification offered is, more or less, that GOF advocates are right and GOF opponents are wrong, so that this rhetoric is justifiable from one side but fallacious when used by the other. A more specific case is, of course, presented, but it is an uncritical summary of pro-GOF arguments that ignores the well-known arguments of critics. The response and the editorial are of no help in deciding which side is right. The editorial misleadingly presents a view that depends logically on a pro-GOF position as though it is an argument that supports a pro-GOF position.

As alluded to in my original comment, much of the GOF debate is about whether the experiments are more likely to cause a pandemic than prevent one. Casadavall appears to have reached a particular conclusion, but critics of GOF experiments would obviously dispute it. They would question the likelihood of a natural H5N1 pandemic, and whether GOF experiments tell us much about this likelihood. Critically, they would question the value of GOF experiments for prevention of such a pandemic, a value that remains purely hypothetical. (In contrast, refraining from performing a GOF experiment assures that this experiment produces no pandemic.) They would point to the return of H1N1 influenza from the laboratory in 1977, and to reports of frequent laboratory accidents with infectious agents, to refute the claim that their fear is purely theoretical.

Would the viruses produced by the H5N1 GOF experiments, if released, be capable of causing human pandemics? If so, then there is a real danger to consider. If not, then these experiments do not provide strong evidence for the likelihood of a natural pandemic, and their general relevance to human pandemics is dubious.

On 2014 Oct 29, Arturo Casadevall commented:

We agree with Joshua L. Cherry that both the pro- and anti-GOF camps have invoked the apocalypse. While pro- and anti-GOF camps invoke the apocalypse in their arguments and emphasize the benefits and risk of such work, respectively, both appeal to fear. However, when it comes to fear there are some significant differences. The fear of naturally caused pandemic comparable to 1918 finds support in the historical record and the available scientific information on influenza virus, including that generated by GOF experiments showing that it has the capacity for mammalian transmissibility. In contrast, fears of a pandemic from resulting from a mishap with a GOF-type of experiment remain a theoretical possibility. We note that such experiments are currently done with a high level of bio-containment and biosafety. Hence, the ‘apocalyptic fallacy’ applies only to use of the apocalypse as a rhetorical device by the anti-GOF camp for the apocalypse is invoked with a vacuum of evidence for this presumed catastrophic event. Given the differences in historically- and theoretically-based fears we do not feel that the use of the apocalypse by pro- and anti-GOF camps as a rhetorical device is comparable.

On 2014 Oct 27, Joshua L Cherry commented:

From the title of this editorial I expected a critique of proponents of controversial gain-of-function (GOF) experiments. Perversely, it levels criticism only at the other side of the debate. According to the editorial, opponents of these experiments have used the possibility of a global pandemic (caused by laboratory-produced virus) as a rhetorical device that is so frightening that it trumps reason. It ignores the fact that proponents of these experiments have always invoked the dangers of a deadly (natural) global pandemic to argue that the experiments are critical for human health.

"In the GOF debate," the editorial states, "the repeated mention of the likelihood of a pandemic is an apocalyptic rhetorical device." Perhaps it is, but who in the debate uses this device? The proponents of GOF experiments clearly do. The specter of a devastating pandemic is central, for example, to Yoshihiro Kawaoka's defense of his experiments. He tells us of the frighteningly high rate of death among confirmed H5N1 infections, and writes that

"Within the past century, 'Spanish' influenza, which stemmed from a virus of avian origin, killed between 20 million and 50 million people. Because H5N1 mutations that confer transmissibility in mammals may emerge in nature, I believe that it would be irresponsible not to study the underlying mechanisms."

Similarly, an Erasmus Medical Center press release about work by Ron Fouchier and colleagues bears the headline "Avian influenza could evolve into dangerous human virus", tells us that "The discovery is important as it could prevent a severe pandemic from occurring", and suggests a 60% death rate for H5N1 infection. A piece by Fouchier and others cites the same death rate data and suggests that the toll of an H5N1 pandemic would exceed that of the H1N1 pandemic of 1918. Many additional examples could be mentioned. This side of the debate repeatedly invokes the danger of a deadly pandemic, and presents GOF experiments as necessary for our rescue from this danger.

(It should be noted that the high death rate for H5N1 infection has been called into question. Some defenders of GOF experiments have used diminished estimates of lethality to downplay the danger of the laboratory-produced viruses, usually failing to note that they would also weaken the original argument for the importance of the experiments.)

It was quite reasonable for critics to ask whether GOF experiments are more likely to cause a global pandemic than prevent one. The critics' discussions of pandemics have generally been no more apocalyptic than those of the proponents, such as those referred to above. The editorial's accusations concerning misleading rhetoric are at best one-sided, and arguably are pointed in exactly the wrong direction.

On 2015 Feb 25, Gerard Ridgway commented:

This work is generally very thorough, but the investigation of discriminant analysis seems rather limited. Only linear and quadratic discriminant analysis are considered, and there is little detail about the methods (for example, no mention of dimensionality reduction or regularisation terms). With up to 180 features, regularised and/or kernel variants of LDA, or other methods such as support vector machines (used in e.g. Ecker C, 2010) might well perform considerably better.

It could be an interesting exercise if the authors would be willing to share their features (to save other researchers from the time-consuming process of running FreeSurfer on many subjects) along with labels for a randomly selected half of the subjects. Other researchers could then submit their predictions for the unlabelled half to Haar et al. for evaluation of the accuracy. I believe the numbers here would be sufficient for a split-half validation to provide meaningful results.

On 2017 Mar 24, Leonid Schneider commented:

I was invited by the journal to submit a letter to the editor to point out some factual inconsistencies in this publication (e.g. patient is dead since 2011, but presented as apparently still alive). Unfortunately, my letter failed peer review. Here it is, together with reviewer reports: https://forbetterscience.com/2017/03/03/my-walles-trachea-transplant-reporting-fails-peer-review/

On 2017 Jul 21, Randi Pechacek commented:

Embriette Hyde wrote a blog about this paper on microBEnet.

On 2016 Aug 23, Ben Goldacre commented:

This trial has the wrong trial registry ID associated with it on PubMed: both in the XML on PubMed, and in the originating journal article. The ID given is NCT0042012. We believe the correct ID, which we have found by hand searching, is NCT00420212.

This comment is being posted as part of the OpenTrials.net project^[1] , an open database threading together all publicly accessible documents and data on each trial, globally. In the course of creating the database, and matching documents and data sources about trials from different locations, we have identified various anomalies in datasets such as PubMed, and in published papers. Alongside documenting the prevalence of problems, we are also attempting to correct these errors and anomalies wherever possible, by feeding back to the originators. We have corrected this data in the OpenTrials.net database; we hope that this trial’s text and metadata can also be corrected at source, in PubMed and in the accompanying paper.

Many thanks,

Jessica Fleminger, Ben Goldacre*

[1] Goldacre, B., Gray, J., 2016. OpenTrials: towards a collaborative open database of all available information on all clinical trials. Trials 17. doi:10.1186/s13063-016-1290-8 PMID: 27056367

* Dr Ben Goldacre BA MA MSc MBBS MRCPsych<br> Senior Clinical Research Fellow<br> ben.goldacre@phc.ox.ac.uk<br> www.ebmDataLab.net<br> Centre for Evidence Based Medicine<br> Department of Primary Care Health Sciences<br> University of Oxford<br> Radcliffe Observatory Quarter<br> Woodstock Road<br> Oxford OX2 6GG

On 2014 Nov 13, Robert Eibl commented:

Dear Readers,

Here is a link to my critical comment on a paper, to which the authors published the above mentioned response: http://scitation.aip.org/content/aip/journal/apl/104/23/10.1063/1.4882182 In my view the authors do not address my fundamental critique very well; a cross-linker just binds any membrane protein covalently to the cantilever, but not all proteins in a membrane are related to cell adhesion. I recommend including a specific cell adhesion receptor, then bound to the cross-linker and repeat the experiment to start measuring real cell adhesion. Pubmed does not include my comment as a reference, although it recently added this response to my comment. This seems to be due to the publisher who only recommends NIH-funded comments and replies to be listed in Pubmed. In contrast, my comment is accurately mentioned in other scientific libraries, for example IEEE Xplore. For those interested in my research field of specific cell adhesion of living cells measured by nanotech methods and on a single-molecule level, I include a list of references; some of my book chapters are also not included in Pubmed.

Best regards, Dr. Robert Eibl

REFERENCES

1. Moy VT et al. , Science (1994)

2. Eibl RH and Moy VT, Atomic force microscopy measurements of protein-ligand interactions on living cells. In: Protein-Ligand Interactions, (Editor: G.Ulrich Nienhaus), Humana Press, Totowa, NJ, U.S.A., pp. 437-448 (2005)

3. Benoit M et al. Nature Cell Biology (2000)

4. Eibl RH and Benoit M, Molecular resolution of cell adhesion forces. IEE - Nanobiotechnology 151(3):128-132 (2004)

5. Eibl RH, Direct force measurements of receptor-ligand interactions on living cells. In: Applied Scanning Probe Methods XII - Characterization. Bhushan B, Fuchs H (Editors), Springer, pp. 1-31, (2009)

6. Eibl RH, Cell adhesion receptors studied by AFM-based single-molecule force spectroscopy. In: Scanning Probe Microscopy in Nanoscience and Nanotechnology 2, Bhushan B. (Editor), Springer, pp.197-215, (2011)

7. Eibl RH, Single-molecule studies of integrins by AFM-based force spectroscopy on living cells. In: Scanning Probe Microscopy in Nanoscience and Nanotechnology 3, Bhushan B. (Editor), Springer, pp.137-169, (2013)

8. Eibl RH, Comment on “A method to measure cellular adhesion utilizing a polymer micro-cantilever” [Appl. Phys. Lett. 103, 123702 (2013)]. Applied Physics Letters, 104, 236103 (2014)

On 2014 Oct 16, Michael Cariaso commented:

The abstract uses 2 nearly identical rs#s, one of which is an error. rs1059111 appears to be correct, rs105911 appears to be a typo

On 2014 Oct 21, George McNamara commented:

This is a nice paper. The abstract refers to using 24 epitope tags (24mer), much of the paper uses a 10mer. Just doing GFP is boring. When I came up with the "Tattletales" (TALE-FPn ... I came up with the idea before sgRNA:Cas9 became popular), I immediately realized that multimerizing FP biosensors. The current paper is the same as my what I refer to as "Binary Tattletales", as in: 1. TALE-(linker-epitope tag)n 2. "binder"-(linker-FP)m with Tattletales being T-cells -- TALE FPs/Biosensors. Since I moved to MD Anderson Cancer Center, the first T now refers to "T-cells and Tumor cells". Likewise T-bow refers to rainbow T-cells and Tumor cells for promoter bashing and otherwise multicolor dots labeling cells (rainbow in homage of course to Brainbow mice etc, and especially to real rainbows). For more on Tattletales, Binary Tattletales, and T-Bow, see http://works.bepress.com/gmcnamara/63 http://works.bepress.com/gmcnamara/42

Giving credit where credit is due: The authors really should have cited the first mammalian cell paper localizing a lot of FPs in one spot (they came 'close' with a Gordon 1997 Cell paper on GFP:LacO in E.coli, but the Tanenbaum paper is all mammalian cells): Robinett et al 1996 JCB http://www.ncbi.nlm.nih.gov/pubmed/8991083 http://jcb.rupress.org/content/135/6/1685.long See their figure 4A. Straight, Robinett et al also published a yeast paper in 1996, http://www.ncbi.nlm.nih.gov/pubmed/8994824 and it would have been useful to cite that.

The PDF download at http://works.bepress.com/gmcnamara/63 has a table of 130 FP biosensors (if you are Laconic about ATeam and Fire, too bad) and an extensive reference list with ZF-FP, TALE-FP, Cas9-FP (the latter from the Weissman group), and more (PUF's and PPR's are RNA binding protein families with structural similarities to TALEs). My favorite name -- besides Tattletales and T-Bow, of course -- is "TALE-Lights" from Yuan, Shermoen, O'Farrell 2014, http://www.ncbi.nlm.nih.gov/pubmed/24556431

On 2015 Aug 17, Alem Matthees commented:

In response to post-publication feedback, I wish to clarify some aspects of the abstract, so there are no further misunderstandings about the scope and content in the full text of this article:

a) Classification and naming issues aside, ME/CFS occurs at all ages, including young children and adolescents. [1] I never intended to suggest otherwise. The statement about patients being almost exclusively of working age was in context of research cohorts, particularly intervention trials which typically exclude patients under 18 years of age and rarely recruit those over 65 years. It is argued that studies consisting of such cohorts should not use normative data from general populations which include the elderly.

b) The physical function subscale of the Short Form 36 health survey (PF SF-36) is discussed because it is a commonly used measure in research and was used in the PACE trial. This article is not a defence of the PACE trial, but uses it to exemplify how the issues described earlier in the article can cause normative data to be misinterpreted or misapplied. Selected details on this issue can be found in an BMJ Rapid Response (open-access) which does not require subscription to view. [2]

c) This article is not meant to be a comprehensive analysis of recovery or case definitions, it is simply a commentary which focuses on using normative data from other comparison groups, one of the issues raised in the review by Adamowicz et al. [3] It explores the appropriate control groups or comparison populations, highlights a problem with using the mean ±1 SD as a threshold if the data does not follow a normal distribution, includes some summary statistics, mentions statistical testing at the group level, and encourages researchers to publish enough information so that others can accurately estimate the functional status of participants. Subjective self-reported measures are important but have potential biases (particularly in nonblinded trials lacking placebo control). Objective measures are also important, particularly when assertions that the intervention is effective at increasing function and activity are contradicted by a range of objective measures. See commentaries by Twisk [4] and others. [5-7]

References

1: Bakken IJ, Tveito K, Gunnes N, Ghaderi S, Stoltenberg C, Trogstad L, Håberg SE, Magnus P. Two age peaks in the incidence of chronic fatigue syndrome/myalgic encephalomyelitis: a population-based registry study from Norway 2008-2012. BMC Med. 2014 Oct 1;12:167. doi: 10.1186/s12916-014-0167-5. PMID 25274261. http://www.ncbi.nlm.nih.gov/pmc/articles/pmid/25274261

2: Matthees A. Re: Tackling fears about exercise is important for ME treatment, analysis indicates. BMJ Rapid Response, 21 January 2015. http://www.bmj.com/content/350/bmj.h227/rr-16

3: Adamowicz JL, Caikauskaite I, Friedberg F. Defining recovery in chronic fatigue syndrome: a critical review. Qual Life Res. 2014 Nov;23(9):2407-16. doi: 10.1007/s11136-014-0705-9. Epub 2014 May 3. PMID: 24791749. http://link.springer.com/article/10.1007/s11136-014-0705-9

4: Twisk FN. A definition of recovery in myalgic encephalomyelitis and chronic fatigue syndrome should be based upon objective measures. Qual Life Res. 2014 Nov;23(9):2417-8. doi: 10.1007/s11136-014-0737-1. Epub 2014 Jun 17. PMID: 24935018. http://link.springer.com/article/10.1007/s11136-014-0737-1

5: Kindlon TP. Objective measures found a lack of improvement for CBT & GET in the PACE Trial: subjective improvements may simply represent response biases or placebo effects in this non-blinded trial. BMJ Rapid Response, 18 January 2015. http://www.bmj.com/content/350/bmj.h227/rr-10

6: Wilshire CE. Re: Tackling fears about exercise is important for ME treatment, analysis indicates. BMJ Rapid Response, 19 January 2015. http://www.bmj.com/content/350/bmj.h227/rr-7

7: Faulkner G. In non-blinded trials, self-report measures could mislead. Lancet Psychiatry. Volume 2, No. 4, e7, April 2015. doi: 10.1016/S2215-0366(15)00089-9 http://www.thelancet.com/journals/lanpsy/article/PIIS2215-0366(15)00089-9/fulltext

On 2015 Apr 04, Arnaud Chiolero MD PhD commented:

A high quality review showing the association between out-of-office BP and target organ damage in children. Much remains to be done

On 2014 Oct 11, Jonathan Eisen commented:

Lots and lots and lots of press for this paper. While thie work seems very important, some of the press is seriously overhyped. Paul Knopfler has a nice overview on his blog: http://www.ipscell.com/2014/10/top-10-takeaways-from-harvard-stem-cell-diabetes-paper/.

On 2014 Oct 11, Jonathan Eisen commented:

The senior author, Doug Melton has posted the authors version of this paper. See http://hsci.harvard.edu/files/hsci/files/pagliuca_et_al_cell_2014.pdf. Thank you Dr. Melton.

On 2014 Dec 17, Sean Ekins commented:

Slides on this are here http://www.slideshare.net/ekinssean/new-target-prediction-and-vizualization-tools-incorporating-open-source-molecular-fingerprints-for-tb-mobile-version-2

On 2014 Dec 17, Sean Ekins commented:

An earlier post on this http://www.collabchem.com/2014/02/20/tb-mobile-version-2-0/ and here it is on app store for free https://itunes.apple.com/us/app/tb-mobile/id567461644?mt=8

On 2014 Dec 15, Sean Ekins commented:

Also added to our list of 2014 papers http://www.collabchem.com/2014/12/15/a-year-in-publications-2014/

On 2014 Dec 02, Sean Ekins commented:

TB Mobile 2 mentioned here http://www.collabchem.com/2014/11/04/app-exposure/

On 2015 May 21, Christopher Southan commented:

See a structural comparison of PDB ligands from different sources including this one http://cdsouthan.blogspot.se/2015/05/will-real-pdb-ligands-please-stand-up.html

On 2014 Oct 12, George Ntoumenopoulos commented:

I completely concur with these authors. In addition the potential impact of ventilatory and patient positioning strategy on the movement of airway secretions also deserves attention. Our recent case report (Physiother Res Int. 2014 Jun;19(2):126-8. doi: 10.1002/pri.1563. Epub 2013 Aug 17. Justification for chest physiotherapy during ultra-protective lung ventilation and extra-corporeal membrane oxygenation: a case study.Cork G1, Barrett N, Ntoumenopoulos G.) highlights the potential failings of current recommendations for secretion clearance. The ultra protective ventilator settings combined with head of bed upright positioning may predispose to secretion retention and warrants further investigation.

On 2016 Apr 02, Daniel Tsin commented:

I will like to mention that : Historically Dr. D.O. Ott performed a ventroscopy appendectomy during vaginal hysterectomy on April 26, 1906 and was published in 1908 : Ott D.O. (1908). Results achieved by the use of direct illumination of the abdominal cavity, colon and urinary bladder during operations and for examination. Russky Vrach 43: 3-11.

Besides our team performed transvaginal appendectomies under the name of Culdolaparoscopy without hysterectomy : Tsin DA, Colombero LT, Mahmood D, Padouvas J, Manolas P.Operative culdolaparoscopy: a new approach combining operative culdoscopy and minilaparoscopy.J Am Assoc Gynecol Laparosc. 2001 Aug;8(3):438-41 .

https://www.ncbi.nlm.nih.gov/pubmed/11509789?dopt=Abstract

On 2014 Oct 30, Christopher Southan commented:

It would be valuable to submit these 326K as a new PubChem source (https://pubchem.ncbi.nlm.nih.gov/)

On 2016 Aug 08, Christopher Southan commented:

Seems a shame if the 10yr momentum has stalled. Any prospect of updating from the declared 2013 statistics?

On 2014 Oct 31, R Y Seedat commented:

Thank you for the positive comments regarding the study.

Lesotho is a different country, with different demographics and burden of disease and a different healthcare system with differences in accessibility, availability of services and referral protocols. It would thus not make sense to combine the data.

I don't think that the calculation of confidence intervals would be a valid statistical test since the study was not based on a population samples but on entire populations. While the incidences calculated are an underestimate, the calculation of confidence intervals would not assist in determining the degree to which the incidence is underestimated.

On 2014 Oct 29, Farrel Buchinsky commented:

Nicely done study and well reported. I have seen several incidence reports but almost always from North America, or Denmark (or another developed country) and never from anywhere in Africa. The best-done North American studies show incidence of about 1 per 100000 or below (around 0.2-1). As the author states, The data described in the Free State and Lesotho is probably an underestimate since pediatric hoarseness (sans dyspnea or stridor) may not be diagnosed amongst those with less access to health care.

• If all the cases from Lesotho are referred to Bloemfontein why not combine the statistics?<br>
• Also, one could provide 95% confidence intervals (Poisson model probably) around the best estimates and see where the estimates from other reports fall within that range.

On 2014 Oct 20, Attila Csordas commented:

associated datasets available via PRIDE/ProteomeXchange: http://www.ebi.ac.uk/pride/archive/simpleSearch?q=PXD001042+PXD001390&submit=Search

On 2014 Oct 21, Bertie Gottgens commented:

One point worth making may be that the studies reported here were presumably done under pathogen-free conditions, and in the absence of predators or any other environmental stresses. "Normal" hemaopoiesis in the real world may therefore have to call on blood production from the traditional 'transplantation HSCs' more often than is suggested here.

On 2016 Jun 14, Christopher Uhde commented:

This study was commented on as the principle subject of two Correspondence articles in Development:

Transcriptional interpretation of Shh morphogen signaling: computational modeling validates empirically established models. Uhde CW, Ericson J. Development. 2016 May 15;143(10):1638-40. doi: 10.1242/dev.120972. No abstract available. PMID: 27190032

and

Mathematical models help explain experimental data. Response to 'Transcriptional interpretation of Shh morphogen signaling: computational modeling validates empirically established models'. Cohen M, Page KM, Perez-Carrasco R, Barnes CP, Briscoe J. Development. 2016 May 15;143(10):1640-3. doi: 10.1242/dev.138461. No abstract available. PMID: 27190033

On 2014 Oct 14, Jonathan Eisen commented:

I would like to point out that I and multiple coauthors published a paper on pooling mitochondrial genomes and shotgun sequencing them for biodiversity analysis in 2000. I believe our paper should have been cited and discussed in this paper. See A case for evolutionary genomics and the comprehensive examination of sequence biodiversity. Pollock DD, Eisen JA, Doggett NA, Cummings MP. Mol Biol Evol. 2000 Dec;17(12):1776-88..

Abstract is pasted below:

Comparative analysis is one of the most powerful methods available for understanding the diverse and complex systems found in biology, but it is often limited by a lack of comprehensive taxonomic sampling. Despite the recent development of powerful genome technologies capable of producing sequence data in large quantities (witness the recently completed first draft of the human genome), there has been relatively little change in how evolutionary studies are conducted. The application of genomic methods to evolutionary biology is a challenge, in part because gene segments from different organisms are manipulated separately, requiring individual purification, cloning, and sequencing. We suggest that a feasible approach to collecting genome-scale data sets for evolutionary biology (i.e., evolutionary genomics) may consist of combination of DNA samples prior to cloning and sequencing, followed by computational reconstruction of the original sequences. This approach will allow the full benefit of automated protocols developed by genome projects to be realized; taxon sampling levels can easily increase to thousands for targeted genomes and genomic regions. Sequence diversity at this level will dramatically improve the quality and accuracy of phylogenetic inference, as well as the accuracy and resolution of comparative evolutionary studies. In particular, it will be possible to make accurate estimates of normal evolution in the context of constant structural and functional constraints (i.e., site-specific substitution probabilities), along with accurate estimates of changes in evolutionary patterns, including pairwise coevolution between sites, adaptive bursts, and changes in selective constraints. These estimates can then be used to understand and predict the effects of protein structure and function on sequence evolution and to predict unknown details of protein structure, function, and functional divergence. In order to demonstrate the practicality of these ideas and the potential benefit for functional genomic analysis, we describe a pilot project we are conducting to simultaneously sequence large numbers of vertebrate mitochondrial genomes.

On 2014 Dec 02, Helen E Benson commented:

The IUPHAR/BPS Guide to PHARMACOLOGY contains curated information on ENaC and ASICs.

On 2015 Mar 27, KLAUS KAESTNER commented:

Dear Dr. Tarlow We are well aware of the fact that a CreER is required for genetic lineage tracing. We tried many times to derive a Foxl1-CreER line. Unfortunately, none of our six founders showed any expression. We will keep trying!

In the case of Foxl1 expression in the liver, there is an additional difficulty to consider. There are no Foxl1 expressing cell in the healthy, uninjured liver. Thus, even a Foxl1-CreER does not help, as it will not label any cell! The Foxl1 promoter becomes activated only AFTER specific types of liver injury.

Note that additional evidence for the bi-lineage potential of Foxl1-Cre marked cells comes from the fact that one can establish clones of cells from a SINGLE Foxl1-Cre/RosaYFP labelled cell, and these can be expanded indefinitely. Thus, these cells are clonogenic. In vitro, these cells can be differentiated towards both the cholangiocyte and hepatocyte lineage.

Note also that we acknowledge the limitations of our current model in the discussion of this paper. Klaus Kaestner

On 2014 Dec 23, Branden David Tarlow commented:

It's unclear why this group has not replicated their results with the Foxl1-Cre with a CreERT2 allele since the original 2009 publication. I agree with a recent review that stated "Experiments using non-inducible Cre lines do not constitute bona fide lineage tracing tools..." (see Lemaigre Hepatology 2014; doi: 10.1002/hep.27659)

On 2015 Dec 30, Vatsalya Vatsalya commented:

SOCIAL SECURITY Fact Sheet: For persons born in 1938 or later, their Social Security benefit may be affected by a provision that raises the age at which full Social Security benefits are payable.

The age for collecting full Social Security retirement benefits will gradually increase from 65 to 67 over a 22-year period beginning in 2000 for those retiring at 62.

Web-link: https://www.ssa.gov/pressoffice/IncRetAge.html

On 2014 Oct 08, Elizabeth Moylan commented:

Thank you for raising this, we are investigating in accordance with COPE guidelines (http://publicationethics.org/).

Elizabeth Moylan, Biology Editor, BioMed Central.

On 2014 Oct 08, Neil Saunders commented:

A substantial fraction of the text in this article has been copied verbatim from an earlier article which it cites. This can be demonstrated by entering the article URLs - http://genomebiology.com/2010/11/3/r25 and http://www.biodatamining.org/content/7/1/15 - into this online tool. Also, the first 10 or so references are identical and in the same order.

Discussion at this Twitter thread includes images which make the similarity very apparent.

On 2015 Jan 29, CREBP Journal Club commented:

This is a very informative and interesting paper, which highlights the utilities and caveats in the use of multiple-indication reviews. The coherent methodology of this article is admirable and we acknowledge that there is a lack of coherent terminology. The term “multiple-indication review” is unambiguous and should be used in future studies. We agree that producers of systematic reviews should consider using this kind of reviews instead of, or in addition to, reviews focusing on a single indication. Important information that we cannot get with traditional methods (single-indication reviews) can be achieved if this methodology is followed. For example, multiple-indication reviews are very useful in the fight against antibiotic resistance. Providing Health Care Professionals and patients with comprehensive information about the risk of adverse effects and the benefit-harm trade-off may reduce their desire for use of antibiotics. Chen et al identified three uses of multiple-indication reviews. However, we propose that the use of this kind of review can be broadly categorised as either:

● To get a better estimate of the effectiveness or harms e.g., 'What are the adverse effects of amoxicillin?' (P* I C H1, H2, …)

● To examine heterogeneity across indications or interventions e.g., 'When are prophylactic antibiotics effective?' (P1, P2, P3, … I C O1), or 'What is the optimum timing of prophylactic antibiotics before any surgery?' (P* I1 I2 I3, … C O1)

(PICO notation: P* = any disease; P1 P2 P3 = set of disease, I1 I2 I3 = set of interventions, C = comparison, O1 = outcome, H = harm)

When undertaking a multiple-indication review much attention has to be paid to the methodologically caveats such as ‘overlapping’ of included reviews; the extra level of complexity (potential heterogeneity in the contributing systematic reviews, in addition to heterogeneity in the primary trials); potential confounders (e.g. methodological quality of included studies and reviews) and risk of bias (e.g. different duration or dose of tested treatment or different control groups). Some of these methodological challenges can be dealt with in the designing of the review and some should be taken into account in the analytical approach. ‘Overlapping’ can be circumvented if only the data from the originals trials are included in the multiple-indication review. However, if the multiple-indication review is based on results from systematic reviews one need to take account of the potential ‘overlapping’ of included studies and we would be very interested in software routines to conduct these reviews - especially in a 2-step frequentist approach. See CREBP Journal Club for more information

On 2014 Dec 08, George Parris commented:

Between 2004 and 2009 I developed a hypothesis for the origin of HIV-1M based on the idea that anti-malarial drugs may have steered the evolution of HIV-1 along a unique trajectory Parris GE, 2004 Parris GE, 2007 Parris GE, 2007. My model deviates substantially from the widely accepted zoonosis model advocated by Sharp and Hahn (2008, 2011). Now, Faria et al. have independently proposed a model of HIV spread based on a detailed conventional analysis of the evolution of the HIV-1M sequence, which is consistent with my model:

Clearly the transfer of SIV to humans in SE Cameroon (about 1876 according to recent analysis Wertheim JO, 2009) substantially predates the MRCA of HIV-1M (1920s). Thus, the original SIV infection in humans lingered for as much as 50 years (1876-1926) and mutated without producing a pandemic while evolving into the HIV-1 clade. Through the 1920s many persons from SE Cameroon with HIV-1 likely visited Leopoldville (Kinshasa) via the Sangha River (see the reports of Louise Pearce concerning African sleeping sickness dating from 1921). The MRCA of HIV-1M subsequently arose in Leopoldville as a result of some unique mutations (not shared with HIV-1O or other HIV-1 groups). The date of this unique transformation has been the subject of several studies and according to Faria et al. (page 60) if subtypes B and C (which obviously evolved later) are eliminated from the dataset, the molecular clock is more appropriately calibrated and the date of the MRCA of HIV-1M settles at 1926. They point out that neither population growth nor genital ulcer disease can explain the "starburst" pattern of divergence of the numerous HIV-1M lines (i.e., few subtypes in the period 1926-1950 followed by many subtypes and sublines in the 1950s and beyond). Finally, they conclude that their findings are consistent with "iatrogenic interventions in Kinshasa...were critical for the emergence of group M." In my papers I describe a specific (well-documented) iatrogenic event in early 1927 in Leopoldville/Kinshasa, which involved a child associated by name and disease (she had sleeping sickness) with SE Cameroon and the Sangha River. It is documented that after being treated with a powerful anti-malarial (pamaquine that affects the white blood cells) she and two other young girls were tested for malaria (blood drawn by syringe) at the same place and same day in sequence.

The substantial delay in the spread of HIV-1M from Leopoldville (from 1927-1937)clarified by Faria et al. is consistent with the virus being in the bodies of children (under 10-years old in 1927). Had the virus been in the adult population (1926-onward) it undoubtedly would have spread geographically much earlier via rail and boat; moreover as a result of commercial sex workers and screening for sleeping sickness (e.g., by Jamot in Cameroon), there would have been many more unique subtypes generated in the period 1926-1940.

It is incredible to me that in spite of the fact that anti-malaria drugs (e.g., chloroquine) are known to affect the replication of HIV (see papers by Savarino A starting in 2001 Savarino A, 2001) that this effect has been universally ignored in analysis of the evolution of HIV. Chloroquine became a daily fact of life in the Congo Basin in the late 1950s, just when the starburst is observed. Moreover, suppression of HIV by chloroquine can explain why HIV was first detected in the US in the 1980s (where chloroquine was not being widely used).

On 2014 Dec 11, Ibrahim Masoodi commented:

This interesting case brings to the fore the multiple complications of SLE, a clinical scenario due to the generation of multiple autoantibodies directed towards the pancreas and the hemopoietic system. Acute vasculitis with its varied clinical presentation is often the commonest cause of abdominal pain in SLE, but acute pancreatitis and acute autoimmune hemolytic anemia, with its consequences, should be kept in mind in a given case of SLE. Further, this case emphasizes the need for close follow-up and proper health education in an SLE patient in order to circumvent any complication.

On 2015 Feb 24, Jonathan S Hausmann commented:

We read with great interest this article by Lukens JR, 2014 on the role of diet in the development of inflammatory bone disease in Pstpip2^cmo mice. They report that mice fed a low-fat diet had elevated levels of pro-IL-1B and developed inflammatory bone disease, a disease which resembles the human autoinflammatory disease chronic multifocal osteomyelitis. However, mice fed a high-fat diet had lower pro-IL-1B levels and did not develop osteomyelitis, despite their genetic predisposition.

The authors conclude that the different outcomes are due to differences in the composition of the diets. However, the diets differed in more than fat content—notably in gluten content. The low fat diet, LabDiet 5013 has wheat as a main ingredient, whereas the high-fat diet, Research Diets Incorporated D12107 contains no wheat and is gluten-free.

Pro-inflammatory effects of gluten have previously been demonstrated in mice. Non-obese diabetic (NOD) mice fed a standard wheat-containing diet had increased levels of inflammatory cytokines, as compared to those maintained on a gluten-free diet during gestation and early life (Hansen CH, 2014). Furthermore, those on gluten-free diets showed changes in the microbiome which conferred protection against the development of diabetes despite their genetic susceptibility.

In patients with familial Mediterranean fever (FMF), another autoinflammatory disease, attempts to modulate the frequency and severity of attacks based on the fat content of the diet have been largely unsuccessful (SOHAR E, 1962). It is now known that the microbiome of patients with FMF differs from that of healthy controls (Ktsoyan ZA, 2013). What has yet to be determined is how diet affects this altered microbiome, and whether a gluten-free diet can promote an anti-inflammatory microbiome and modulate the clinical course of FMF and other autoinflammatory diseases.

The question whether fat or gluten were responsible for the observed changes does not change the remarkable findings by Lukens JR, 2014 that diet can change the expression of this genetic autoinflammatory disease in mice by modulating the microbiome. However, when considering these results in developing possible treatments, it will be important to clearly determine the ingredients that changed the microbiome for the better.

On 2016 Feb 24, Lily Chu commented:

Above link is broken. See here for new link:

http://iacfsme.org/ME-CFS-Primer-Education/Bulletins/2009/Severe-ME-CFS-in-Adults-A-report-from-the-CH-(1).aspx

On 2014 Oct 21, Lily Chu commented:

The results of this study confirm those of an earlier study by Pheby et al. in 2009. That study also noted two onset peaks, one from ages 11-20 and another from 31-40.

Pheby's article can be accessed online here: http://www.iacfsme.org/BULLETINWINTER200910/Winter0910PhebySneddonHeinrichCHROMEReport/tabid/413/Default.aspx

On 2015 May 02, Arthur Yin Fan commented:

The original trial by Hinman's has many methodology flaws, please read the comments under the original trial report.

On 2014 Nov 08, Qin-hong Zhang commented:

None

On 2014 Oct 04, Ryan Radecki commented:

Post-publication commentary:

"ARISE, and Cast Off the Shackles of EGDT"

The sound you hear is a sigh of relief from Emergency Physicians and intensivists regarding the outcomes of the Australasian Resuscitation in Sepsis Evaluation (ARISE).

As ProCESS suggested, and as many have suspected all along, it seemed the critical intervention from Early Goal-Directed Therapy was the early part – and less the SCO2 monitoring and active management of physiologic parameters using dobutamine and blood transfusion. Now, we have a second study, in addition to ProCESS, supporting the same general conclusions....

http://www.emlitofnote.com/2014/10/arise-and-cast-off-shackles-of-egdt.html

On 2016 Jun 07, Arthur Yin Fan commented:

Acupuncture is Effective for Chronic Knee Pain: A Reanalysis of the Australian Acupuncture Trial. Altern Ther Health Med. 2016 Mar;22(3):32-6. Yin Fan A, Zhou K, Gu S, Ming Li Y. Abstract Context • In the October 2014 issue of the Journal of the American Medical Association (JAMA), Hinman et al published the results of an Australian clinical trial on acupuncture in a paper entitled "Acupuncture for Chronic Knee Pain: A Randomized Clinical Trial" (JAMA report), in which they concluded that neither acupuncture nor laser acupuncture had any greater effects than sham laser acupuncture for pain or function for patients aged 50 y and older with moderate-to-severe knee pain. That study has been criticized extensively by international scholars for its validity because serious methodological flaws existed throughout the study's design, implementation, and conclusions. Objective • The current study intended to re-examine the prior study's conclusions about the efficacy of acupuncture for chronic knee pain. Design • The current research team performed a reanalysis of relevant data from the JAMA report. Intervention • The original study included 4 groups: (1) an acupuncture group, which received needle acupuncture, inferred by the current authors to have been set up to be a positive control in the original study; (2) a laser acupuncture group, which received laser acupuncture; (3) a sham laser acupuncture group, which received sham laser acupuncture and acted as the negative controls for the laser acupuncture intervention; and (4) a control group, which received conventional care but no acupuncture or laser treatments. The study lasted 12 wk. Outcome Measures • The measures included evaluations in the following areas: (1) poststudy modifications-an evaluation of the consistency of the JAMA report with the study's intentions as identified for a grant that was originally approved and funded by the Australian National Health and Medical Research Council (NHMRC) in 2009, as indicated in the study's trial registration, and as compared with the published protocols and to the study's originally stated objectives; (2) high heterogeneity-an assessment of the heterogeneity among the 4 groups for the overall outcome related to pain; (3) ineffectiveness of laser acupuncture-an analysis of laser acupuncture's efficacy for chronic knee pain as stated in the JAMA report, using effect size (ES); (4) effectiveness of acupuncture-a reanalysis of acupuncture's efficacy for chronic knee pain in comparison with the original analysis in the JAMA report, using ES; and (5) acupuncture after data adjustment-a new analysis of acupuncture's efficacy for chronic knee pain using data from the original study that was discussed in the JAMA report, using ES, with an estimation after data adjustment and elimination of the dilution effect of the Zelen design. Results • Contrary to a general impression that acupuncture was the focus, laser acupuncture was the primary intervention tested in the actual study, "Laser Acupuncture in Patients With Chronic Knee Pain: A Randomized, Placebo Controlled Trial." The study discussed in the JAMA report was neither a truly randomized, controlled trial (RCT) for acupuncture nor was it an appropriately designed, randomized study in general. High heterogeneity was found among its groups in the evaluation of overall pain in patients. Both the ES of 0.60 that had been set by Hinman et al for the minimal clinically important difference (MCID) and the resulting interpretation of results in the JAMA report were not appropriate. Using the original study's criteria of efficacy, the reanalysis has confirmed that the laser acupuncture was not effective, whereas the acupuncture was found to be moderately effective for chronic knee pain (P < .05) for both overall pain and function at 12 wk, with an ES of 0.58, or after the adjustment of the data, with an ES of 0.67. Conclusions • The JAMA study was neither a conventional RCT nor an appropriately randomized trial, and its results are probably invalid. The ES of 0.60 for the MCID that was used in the JAMA study and the resulting explanation were not appropriate. Even with an ES of 0.60 for the MCID, acupuncture remained effective after data adjustment. Consequently, compared with conventional care, acupuncture treatment was found to be moderately effective for chronic knee pain in patients aged 50 y and older. PMID: 27228270 [PubMed - in process]

On 2015 May 30, Arthur Yin Fan commented:

The sample size calculation is inaccurate in this study. http://www.jcimjournal.com/articles/publishArticles/pdf/S2095-4964(15)60184-4.pdf

It should be 773, instead of 282. Hinman described she considered the factors of multiple groups' comparison, intra-therapies,however, she did not.

See the step by step sample size calculation detail in: The methodology flaws in Hinman’s acupuncture clinical trial, Part III: Sample size calculation. http://www.jcimjournal.com/articles/publishArticles/pdf/S2095-4964(15)60184-4.pdf

On 2015 May 29, Hongjian He commented:

One our main criticisms of this publication was that there was no detailed reporting of number of needles used per treatment, acupuncture sites targeted per treatment, and specific treatment duration and electric stimulation for each patient. Dr. Hinman responded by stating:

"Dr. He suggests lack of acupuncture standardization, treatment infrequency, and no electrical stimulation may explain our findings. However, when comparing acupuncture with sham treatment, a meta-analysis1 found no evidence that needle number or placement; use of electrical stimulation; or number, frequency, or duration of treatments influence acupuncture outcomes"

We looked at the article Dr. Hinman cited (MacPherson et al., PLoS 2013) and she did not do a good job of reading it. They showed that number of needles used per treatment was statistically significantly correlated with effect size. The electrical stimulation had a significantly stronger effect. She incorrectly summarized the study results. Therefore, Dr. Hinman’s statement that needle numbers, length of treatment and electric stimulation have no effect on acupuncture outcomes is incorrect. We still await a sufficient explanation for why these specifics were not reported in her study.

On 2016 Apr 09, Leigh Jackson commented:

Wit et al. conducted a pragmatic trial whose design had glaring weaknesses.

There was no sham control. There was no blinding. Control was usual care inviting negative bias. The non-randomized self-selecting cohort invited positive bias. Consort was not followed. Patients had individualised treatment creating a chaotic heterogeneity of data.

Wit et al. to Hinman et al. is as chalk to cheese. The difference in their results might be due to low dosage levels in Hinman et al. It might be due to high levels of bias in Wit et al.

On 2015 Apr 27, Qin-hong Zhang commented:

Acupuncture treatment for chronic knee pain: study by Hinman et al underestimates acupuncture efficacy

http://aim.bmj.com/content/33/2/170.full.pdf+html

Dr Hinman and colleagues [1] completed a Zelen-design clinical trial for acupuncture for chronic knee pain patients and concluded that neither laser nor needle acupuncture conferred benefit over sham for pain or function in patients older than 50 years with moderate or severe chronic knee pain. We disagree with authors because they failed to use the most effective acupuncture regimen in their trial.

We consider that their treatment regimen is inferior for the following reasons. First, the dosage of acupuncture is far from adequate. The protocol specified the acupuncture intervention as a twenty minute treatment once or twice weekly for 12 weeks, with 8 to 12 sessions in total permitted [1]. Treatment compliance was not reported. Even with an assumption of full compliance, the study participants received only 0.67 to 1.0 session weekly with a total 160 to 240 minutes in 12 weeks. This was below the treatment regimen in the trial by Witt, et al. [2], in which participants received 12 sessions of 30 min duration, administered over 8 weeks, i.e., average 1.5 sessions weekly, and 360 minutes in total for only 8 weeks. It is worthwhile to point out that Witt, et al. had opposite conclusions. Second, the study protocol did not require deqi (a renowned acupuncture sensation), which is profoundly regarded as a prerequisite of a preferable acupuncture treatment efficacy [3]. Third, the paper did not follow the CONSORT statement of acupuncture [4] that requires details of needling, such as needle manipulation, depth of needle insertion, and points selected unilateral, bilateral or both. Fourth, the dose of laser acupuncture was 0.2J per acupuncture point, which was considered too low to achieve a clinical effect [5]. Previous study suggested that the minimum dose should probably be 0.5 J/point [5] . Thus it is difficult to evaluate if effective treatment regimen was compared against the Sham.

Because the efficacy of acupuncture therapy depends on the dose and deqi of acupuncture intervention, it is premature to us to reach the conclusion that acupuncture is not effective to treatment osteoarthritis pain of the knee.

REFERENCES

1. Hinman RS, McCrory P, Pirotta M, et al. Acupuncture for Chronic Knee Pain A Randomized Clinical Trial. JAMA. 2014;312(13):1313-1322.

2. Witt C, Brinkhaus B, Jena S, et al. Acupuncture in patients with osteoarthritis of the knee: a randomised trial. Lancet. 2005;366(9480):136-143.

3. Shi GX, Yang XM, Liu CZ, et al. Factors contributing to therapeutic effects evaluated in acupuncture clinical trials. Trials. 2012; 13:42.

4. MacPherson H, Altman DG, Hammerschlag R, et al; STRICTA Revision Group. Revised STandards for Reporting Interventions in Clinical Trials of Acupuncture (STRICTA): extending the CONSORT statement. PLoS Med. 2010;7(6):e1000261.

5. Baxter GD. Laser acupuncture: effectiveness depends upon dosage. Acupunct Med. 2009;27(3): 92.

On 2015 Apr 16, Arthur Yin Fan commented:

The methodology flaws in Hinman’s acupuncture clinical trial, Part II: Zelen design and effectiveness dilutions: http://www.jcimjournal.com/articles/publishArticles/pdf/S2095-4964(15)60172-8.pdf

Hinman and colleagues concluded that “in patients older than 50 years with moderate or severe chronic knee pain, neither laser nor needle acupuncture conferred benefit over sham for pain or function. Our findings do not support acupuncture for these patients.” As pointed out in my former article (The methodology flaws in Hinman’s acupuncture clinical trial, Part I: Design and results interpretation. J Integr Med. 2015; 13(2): 65–68.), there were serious flaws in the trial design and statistics, as well as in the interpretation of the results. In here I address problems in the Zelen design used by them -Using Zelen design in this study does cause biases.

1 High drop-out rate:

The drop-out rates were 2.82% (2/71) in the control group; 22.86% (16/70) in the acupuncture group; 18.31% (13/71) in the laser acupuncture group; and 22.86% (16/70) for the sham laser acupuncture group. According to the acceptable standards for an RCT, dropout rates less than 10% are acceptable, drop-out rates between 10% and 20% mean that the resulting data quality is poor, and drop-out rates of more than 20% mean that the data quality is considered very poor and should not be used in analysis. In this trial analysis, the data quality in the acupuncture and sham laser acupuncture groups are very poor as the drop-out rates are over 20%; the authors should not have directly used them in any statistical analysis, unless they had re-adjusted and re-balanced the sample among the groups during the study. As outlined by the National Institutes of Health, if there is a differential drop-out rate of 15% or higher between study arms, such as between the control group and the treatment group in this clinical trial, then there is a very high potential for bias. This is a flaw that can decrease the quality of the study results.

2 The effectiveness in intervention groups was diluted by various factors

The dilution rates should then be 21.87% in the laser acupuncture group, 13.80% in the sham laser acupuncture group, and 31.27% in the acupuncture group (the dilution rate calculations were shown in Tables 1–3). The dilution rate was very significant in the acupuncture group, which causes the effectiveness to be undervalued in the acupuncture group, by almost 1/3.

The effective significance was masked by limited sample size due to the Zelen design of this study.

3.The sample size calculation in this study is questionable.Too small.

4 Conclusion

The effectiveness of the acupuncture group was diluted 31.27%, and its drop-out rate was 22.86%, much higher than that of the other groups in Hinman’s clinical trial, which constitutes major flaws in how this study is analyzed and interpreted[8]. Based on the bias of Zelen design used in the study, and incorrect sample size calculation, the conclusions drawn from this study are of poor quality, inaccurate, and invalid.

http://www.jcimjournal.com/articles/publishArticles/pdf/S2095-4964(15)60172-8.pdf

On 2015 Apr 11, Arthur Yin Fan commented:

The sham laser acupuncture is not a valid negative control for acupuncture

I agree acupuncture should have a real sham control in a vigorous RCT; however, in Hinman's acupuncture RCT, the sham laser acupuncture is only fit to the laser acupuncture, not to real acupuncture. Because Acupuncture and Sham laser acupuncture, these two interventions do not have comparability in both characteristics and form (i.e., not matched). Furthermore, there was no blinding method performed between these two groups-both the patients and the administrators who performed the interventions knew the difference between the groups, such as needling acupuncture and sham laser acupuncture.

On 2015 Apr 11, Arthur Yin Fan commented:

There is a crucial mistake in interpreting the Hypothesis testing - What means? if P>0.05.

Hinman said :"in......chronic knee pain, neither laser nor needle acupuncture conferred benefit over sham for pain or function (Dr. Fan notes: Her statement was based on P>0.05). Our findings do not support acupuncture for these patients”

From the perspective of hypothesis testing in Statistics, if acupuncture has better results and with significant difference over the primary control (no-treatment group), p<0.05, we can conclude that “acupuncture is effective”- no matter what the result get from the comparing to the secondary control, such as “sham laser acupuncture”, but Hinman intentionally does not report this effectiveness in her conclusion; if acupuncture has better results over “laser acupuncture” and “sham laser acupuncture”, without significant in statistics, p>0.05, we can conclude that “acupuncture is better than the laser acupuncture, and sham laser acupuncture, but need more studies to confirm”. We can’t conclude that “acupuncture is not effective” because that there are no significant difference in statistics between acupuncture and “laser acupuncture”, or between acupuncture and “sham acupuncture” does not mean there is no difference between these treatments clinically. Hinman et al mis-interprets the results and violates the basic principle of Statistics.

On 2015 Apr 11, Arthur Yin Fan commented:

Hinman's acupuncture RCT has too many methodology flaws and misleading

As an independent researcher and practitioner in Acupuncture and Chinese medicine for thirty years, I strongly disagrees with Hinman's conclusion, as there were serious flaws in the trial design, the statistical analysis of the data and in the interpretation of the results of this study. I published a commentary recently [Fan A. The methodology flaws in Hinman’s acupuncture clinical trial, Part I: Design and results interpretation. J Integr Med. 2015; 13(2): 65–68.] http://www.jcimjournal.com/articles/publishArticles/pdf/S2095-4964(15)60170-4.pdf

The major concerns are:

(1)There is a major mistake in the primary testing factor in this RCT: the laser acupuncture should be the primary testing factor, not the needle acupuncture;

(2)The interpretation of the results was misleading;

(3)The “under-dosed” acupuncture treatments diluted the potential real effectiveness of acupuncture;

(4)Laser acupuncture and acupuncture would be effective in Hinman’s RCT, if the statistics were re-analyzed after re-adjusting the data.

(5)It is improper to test two different testing factors in one RCT with so small sample size;

(6)Laser acupuncture is not one kind of acupuncture, the author intentionally mixes it with acupuncture;

(7)Acupuncture did have significant effectiveness (p<0.05 in week 12), compared to the control (this is a primary control). However, the author intentionally does not interpreter this important result into the conclusion, instead, she concludes acupuncture is not effective and says her findings do not support acupuncture for patients.

I feel the author, somehow, intentionally misleads readers by testing acupuncture as a major intervention in this RCT-There was no significance between the positive control and the naïve control (i.e., acupuncture and control groups). Therefore, we can only conclude that the positive control, acupuncture was under-dosed or the study was otherwise flawed. That the positive control shows significance is a basic sign of the success of a clinical trial. From this perspective, Hinman’s trial was a failed clinical trial for laser acupuncture. As it would be unethical to publish an astonishing article, with a group of almost scrapped data and confusing logic, that misleads the readers, including the general public, medical society and policy makers, the researchers should have re-adjusted or re-designed their study instead of publishing it.

On 2015 Jun 14, Arthur Yin Fan commented:

Meridians could be tested by physics method, they are low electric resistance lines.

On 2015 Jun 11, David Keller commented:

It is no longer accurate to label evidence-based science as "Western"

The scientific method is the most powerful tool ever developed for the advancement of humanity, and science has, in turn, been advanced by humans from all cultural backgrounds.

Ancient societies all developed superstition-based pseudo-sciences, which are incapable of predicting natural phenomena or truly understanding them. Astrology was a vain attempt to rationalize the movements of the planets, utterly useless until it was replaced by the science of astronomy. Similarly, alchemy led nowhere until it was transmuted by the scientific method into chemistry. Homeopathy is based on false concepts involving dilution, and yields "medicines" which are nearly pure placebo. Chiropractic medicine, in addition to being based on disproved concepts, can be downright dangerous. PubMed has documented many cases of carotid and vertebral artery dissections caused by chiropractic neck manipulations (search on "artery dissection chiropractor").

The National Institutes of Health were founded to advance the scientific method in pursuing solutions to the health problems which plague mankind. Superstition-based folk remedies should be investigated when there is a chance that ancient peoples may have stumbled on a real medicine - such as aspirin from willow bark or digitalis from the foxglove plant. The active ingredient should be isolated, synthesized, tested and meta-analyzed until it is thoroughly understood, using the scientific method.

Where in the human body can we find anatomical evidence of a meridian, or "chee"? Acupuncture is based on complex and detailed theories, but theories must be tested and proved in the material world, in a reproducible fashion. If acupuncture is incapable of demonstrating significant pain relief in a carefully conducted clinical study, perhaps its adherents should reexamine their beliefs rather than invoking ever more arcane and obscure objections to the study design.

On 2015 May 30, Arthur Yin Fan commented:

Western medicine more focuses on structure, and Eastern medicine more on function. So, when explaining a specific issue, for example, a disc hernia induced lower back pain, even sciatica, western medicine will say the nerve gets pinched, however, eastern medicine will says the Bladder meridian Qi stagnant. Both are correct. Just because using different perspective, and using different language.

What is the meridian? just a functional manifestation of the never. Nerve is the structure, the meridian is the function.

Quite simple.

On 2014 Nov 13, David Keller commented:

Should medical doctors recommend acupuncture to patients with chronic knee pain?

Here is a brief description of acupuncture: "In traditional Chinese medicine, certain body systems, called meridians, are thought to regulate body function through the normal flow of energy [this energy is spelled "qi" but pronounced "chee"] from one part to another. Disturbances in this flow are thought to cause disease. Acupressure and acupuncture techniques are believed by practitioners of traditional Chinese medicine to cure disease by restoring this flow."(1) Without physiological or anatomical evidence of the existence of meridians or qi, acupuncture should be viewed as similar to chiropractic or homeopathic therapies, which lack scientifically credible theoretical mechanisms of action. This study found no significant clinical benefit of acupuncture for chronic knee pain. Therefore, I will not begin referring patients with chronic knee pain for acupuncture. I would like to see this study repeated for chronic pain syndromes of other body regions. If the null result is confirmed in all such regions, acupuncture will have to be classified as an expensive and complex placebo. Funding for research on debunked alternative therapies should be diverted to investigators conducting basic or clinical research of a more scientific nature.

Reference

1: Wang W and Wu S. Treating Pain With Acupuncture. JAMA 2014;312(13):1365.

On 2014 Nov 07, Qin-hong Zhang commented:

None

On 2016 Apr 03, Leigh Jackson commented:

Laser acupuncture provides a means for double blinding. This is a useful methodology for acupuncture trials where double blinding has always been the weakest link.

On 2015 May 30, Arthur Yin Fan commented:

This study used a wrong sham for acupuncture, so the conclusion is not reliable.

On 2014 Oct 08, David Keller commented:

Acupuncture trials have difficulty addressing the placebo effect and blinding

Acupuncture has been challenging to test in a true double-blinded and placebo-controlled fashion. Skilled practitioners of acupuncture have invested years in their training and licensing, and may unintentionally transmit signals of their own strong belief in its purported beneficial effects, augmenting the placebo effect. Simply poking needles into a patient at randomly chosen points could have a strong placebo effect if done with an air of compassion and clinical authority.

It has been argued that sham acupuncture performed as placebo therapy in clinical trials may accidentally stimulate a true acupuncture point, and thereby reduce the apparent benefit of acupuncture in the intervention subjects compared to controls. The use of sham laser acupuncture seems to address that objection.

The arguments and criticisms of acupuncture trials will continue. More good evidence, such as the findings of this study, will be required before we can conclude that acupuncture is of no more benefit for relieving pain than placebo. Better understanding of the biological basis of pain is needed, to develop new and truly effective analgesic therapies.

On 2014 Oct 13, David Keller commented:

Will mapping of all neural circuits increase understanding of the brain in meaningful ways?

The Human Brain Mapping Initiative (HBMI) is a government-mandated research program with the goal of systematically mapping the neural circuitry of the entire human brain. Will this large-scale fiat-funded project result in commensurate advances in neuroscience, or will it divert scarce funds away from smaller, innovative research projects with the potential for breakthroughs in diverse realms? Are we trading away the prospect of new fundamental discoveries, in order to accumulate terabytes of minimally meaningful mapping data in a routine and mechanistic fashion? The brain circuit mapping study by Fox et al provides interesting and relevant findings, much to the credit of the overall Brain Initiative.

The Human Brain Mapping Initiative recalls prior large-scale government-mandated projects, such as the effort to put a man on the moon, and the Human Genome Sequencing Project. The moon shots consumed funding which could have driven great progress in more crucial areas of discovery than lunar geology and micro-gravity ant farming. And, while the Genome Project did yield important information quickly, it has been criticized for amassing huge amounts of expensive data before they were needed, when the same information would have accumulated naturally over the years during the course of other investigations, and at less expense. Supporters have replied that the Genome Project itself drove down the cost of DNA mapping, due to the large market for sequencing equipment it created and funded. Similar arguments can probably be made both to attack and to defend the Brain Mapping Project. Only time, and further results, will settle these questions.

On 2015 Sep 17, Ghassan El-Baalbaki commented:

authors of this comment are: Loose, T.^1,3 , Ashrah, L.^1, Romero, M.^1, Bégin, J.¹ and El-Baalbaki, G.^1,2 Affiliations, Université du Québec À Montréal^1, McGill University^2, Université de Nantes³ .

In this study, the authors examine the efficacy of Acceptance and Commitment Therapy (ACT) in treating partner aggression. After entering into communication with the first author, she confirmed that this study corresponds to her doctoral thesis (Zarling, 2013). We read her thesis in order to gain further knowledge about the published study, and we found two important aspects of the work that need to be brought into light: 1) we find it difficult to conclude that partner aggression was specifically reduced, because of threats to internal validity. 2) There seems to be inaccuracies in their statistical calculations.

The primary objective was to demonstrate that ACT reduces intimate partner aggression better than the placebo control group. The authors strongly suggest that ACT was indeed able to do so. However, several points call this statement into question.

First of all, it is worth noting that partner aggression is never specifically assessed. One may think that the Conflict Tactics Scales-2-Physical Assault Scale (CTS-2) did assess this construct (Straus, Hamby, Boney-McCoy & Sugarman, 1996), but when taking into account the exact content of the administered questionnaire that was figured in her thesis, it can be seen that the instructions were modified in order to include aggression towards “people we care about” (Zarling, 2013, p.132) instead of specifically towards a partner. This modification is not mentioned in the published article. Hence, participants could have hypothetically committed aggressive acts exclusively towards a non-partner, such as a child or a friend. It is thus hard to conclude that aggressive behavior, specifically among couples, decreased as a result of therapy.

Furthermore, after modifying the instructions, one of the items even became incoherent. More specifically, when taking into the modified instructions of the questionnaire, the item stated “When upset or in a disagreement with someone you care about … have you became angry enough to frighten your partner?” (italics added)(Zarling, 2013, p. 132). It seems that this question was destined to ask if the participant became angry enough to frighten someone that they « care about » and not their « partner ». For example, if the participants were (or became) single, they could have interpreted this question as if they were angry enough to frighten someone that is not necessarily their partner. It is unclear how the authors wanted the participants to interpret this item. The way that the construct of partner aggression was assessed calls into question the internal validity of the study.

It remains questionable why the authors neglected to assess attrition of couples during the study. Over the course of the study, it is possible that aggressive behavior decreased more in the ACT group as a result of more couples breaking up in this treatment condition than in the placebo control group. Even if the authors carried out both completer analysis and intent to treat analysis and the results did not differ on measured variables, the attrition of couples was not measured. Hence, it is impossible to know if the groups differed on partner attrition. It remains possible that participants continued to aggress former partners, but again this cannot be asserted simply because it was not measured. Assessing partner attrition would have helped neutralize a pertinent threat to internal validity. Hence, internal validity was limited by not taking into account partner attrition.

Lastly, it is worth bringing up that the first author and two other investigators co-led all administered interventions (p. 201). Even if the authors state this in their article, after looking at the detailed explanation of the recruitment process of investigators, this choice does not seem to be justified. It seems that the first author was able to recruit investigators at no extra cost and it is questionable as to why she did not recruit three instead of two, thus avoiding a potential threat to internal validity. The explanation provided in her thesis, but not the article, also states that all the manipulation checks were carried out by the first author and supervised by the second and third authors (Zarling, 2013, p.61). Because of the implication of the authors in the experimental procedure, the results are potentially attributable to the experimenter expectancy effect. This is another threat to internal validity that evokes the tendency for researchers to unintentionally bias the results of their investigations in accord with their hypotheses. In taking into account the previously stated points, it seems difficult to conclude that this study shows that ACT specifically reduces partner aggression better than a placebo control group.

On another note, the statistical validity of the study can be questioned:

Throughout the results section of the paper (pp. 205–207), the authors report many beta values (β) for intercepts and slopes. If these are indeed beta values, they should vary between approximately 1 and -1 (Cohen, Cohen, West & Aiken, 2002). However in many cases, reported values are outside of this range. For example the authors state “The ACT participants also reported significantly less psychological aggression, β = 2.05, SE = 0.71, t(97) = 8.33, p < .001, and physical aggression, β = 2.21, SE = 0.65, t(97) = 8.19, p < .001, at the 6-month follow-up assessment” (p. 8). This leads us to think that the authors are actually reporting non-standardized coefficients (B) because B values can vary outside of this range. However, if this is the case, then the t values reported become incoherent. t values should equal B/(2SE) (we retained the value of 2 after rounding up from 1.96 which corresponds to 95% of a normal distribution (Christensen, 1986)), but reported t values do not correspond to this calculation. For example, let’s take another look at the previous citation and assume that it actually refers to B values. According to our calculations, t = B/(2SE) = 2.05/(2*0.71)=1.44, but the authors report that t = 8.33, p < .001. More simply stated,if the authors are indeed reporting β values, then many of the β values seem to be impossible (outside of the ± 1 range), but if they are reporting B values, the t values seem to be impossible according to our calculations. We were wondering how these seemingly paradoxical results could be explained.

In conclusion, even if this research topic is of great interest, two main points should to be taken into account. First of all, it is hard to state that ACT is an effective treatment to reduce partner aggression because 1) partner aggression was never specifically assessed 2) partner attrition was not measured 3) of potential bias due to the experimenter expectancy effect. Secondly, it is hard to draw conclusions from the statistics figured in the study because 1) if β values were indeed being reported, then they lie outside of the ± 1 range and 2) if B values were actually reported, then the t values become incoherent.

References

Christensen, H. B. (1986). La statistique: démarche pédagogique programmée. Boucherville: Gaëtan Morin.

Cohen, J., Cohen P., West, S. G. et Aiken, L. S. (2002). Applied multiple regression/correlation analysis for the behavioral sciences. New York: Routledge.

Straus, M. A., Hamby, S. L., Boney-McCoy, S., & Sugarman, D. B. (1996). The revised conflict tactics scales (CTS2): Development and preliminary psychometric data. Journal of Family Issues, 17, 283–316.

Zarling, A. (2013). A preliminary trial of ACT skills training for aggressive behavior. University of Iowa.

On 2014 Oct 02, David Keller commented:

Quicker-and-sicker discharges inevitably result in increased hospital re-admissions

Medicare and many private insurers pay hospitals a fixed amount to treat a given diagnosis, regardless of length of stay, which creates a financial incentive to discharge patients as soon as possible. Targets for shorter lengths of stay are set by hospital managements and are incorporated into the performance reviews of employed hospitalist physicians. With hospitalists pushing the envelope to get patients discharged quicker, an increase in the number of "bounce-back" re-admissions is inevitable, due to inadequate treatment, recrudescence of illness, or the lack of resources or support needed to complete recovery at home. To discourage premature discharges, financial penalties for "bounce-back" re-admissions were created by Medicare and other payers. Of course, these penalties are also incorporated into the performance reviews of employed physicians, and create a disincentive to readmit patients who were recently discharged. But, if the patient presents for treatment at a different hospital, the readmission penalty applies against the first hospital to treat and discharge the patient, not the second hospital. Thus, recently-discharged patients who are still too sick to complete their recovery at home may find it easier to be readmitted at a new hospital for inpatient treatment, where they will be evaluated by physicians who do not have a disincentive to re-admit them. Switching hospitals can make it easier for a still-sick patient to get re-hospitalized, but it also results in discontinuities of care and resulting inefficiencies, because records need to be transferred, new doctors need to learn about the patient, etc. This all just demonstrates that for every action to reduce costs by penalizing providers, there are unintended consequences which can harm patients.

On 2014 Oct 12, Amanda Capes-Davis commented:

Cell lines derived from ductal carcinoma in situ are certainly needed. Many thanks to the authors for including an STR profile for ETCC001 as part of their work. Did the authors test the other derivatives that were transfected with hTERT? It would be worthwhile testing all derivatives, just to confirm that they come from the same donor.

On 2014 Oct 18, stephane burtey commented:

It is interesting to remind that pioglitazone the drug use in the experiments is associated with an increased risk of bladder cancer (http://onlinelibrary.wiley.com/doi/10.1111/bcp.12306/abstract;jsessionid=0BF19A6B1135280E0C6DAAB5F9A2BF21.f02t03) and was withdrawn in France for this reason. It could be very interesting that the authors discuss this strange fact. Why imagine to use a drug known to cause be associated with an increase risk of cancer to treat cancer?

On 2016 Apr 15, Amanda Capes-Davis commented:

This article has now been retracted. See http://www.ncbi.nlm.nih.gov/pubmed/26909548 for the retraction notice and further comment.

On 2014 Oct 15, Amanda Capes-Davis commented:

KB is not an OSCC cell line - KB cells are actually HeLa, from cervical adenocarcinoma. Cross-contamination of KB was discovered by Stanley Gartler back in 1967. Unfortunately, KB is still widely used as a model for oral cancer. Cross-contaminated cell lines are extensively used in the scientific literature, with many scientists not aware of this important problem.

A list of cross-contaminated or otherwise misidentified cell lines can be found at http://iclac.org/databases/cross-contaminations/.

On 2015 Aug 01, James C Coyne commented:

Discussion of recovery patterns and schizophrenia will be facilitated when these authors finally published the overdue results of Klingberg, S., Wittorf, A., Meisner, C., Wölwer, W., Wiedemann, G., Herrlich, J., ... & Buchkremer, G. (2010). Cognitive behavioural therapy versus supportive therapy for persistent positive symptoms in psychotic disorders: The POSITIVE Study, a multicenter, prospective, single-blind, randomised controlled clinical trial. Trials, 11(1), 123. <pubmed/21190574>. This represents the largest clinical trial of cognitive behavior therapy for psychosis ever undertaken.

On 2016 Jun 03, Ewen Gallagher commented:

This work analyses the MEKK1 (encoded by Map3k1) PHD motif genetically and by protein array substrate screening. Shows an important role for the MEKK1 PHD motif in TGF-β cytokine signaling and demonstrates how PHD ubiquitination can influence TAK1 signal transduction. Shows genetically by knockin mutation (Map3k1^mPHD allele) that MEKK1 is important for stem cell MAPK signaling, differentiation and tumorigenesis. Reports phenotypes of Map3k1^mPHD/+ mice. A review is also published for this work (1). Other recent works looking at MEKK1 activation by cytokines and hyperosmotic stress (2-3). A short review is available relating to this work (4).

Related work. (1). Suddason T, Gallagher E. A RING to rule them all? Insights into the Map3k1 PHD motif provide a new mechanistic understanding into the diverse roles of Map3k1. Cell Death Differ 2015. (2). Matsuzawa A, Tseng PH, Vallabhapurapu S, Luo JL, Zhang W, Wang H, Vignali DA, Gallagher E, Karin M. Essential cytoplasmic translocation of a cytokine receptor-assembled signaling complex. Science 2008; 321:663-8. (3). Steed E, Elbediwy A, Vacca B, Dupasquier S, Hemkemeyer SA, Suddason T, Costa AC, Beaudry JB, Zihni C, Gallagher E, et al. MarvelD3 couples tight junctions to the MEKK1-JNK pathway to regulate cell behavior and survival. The Journal of cell biology 2014; 204:821-38. (4). Gallagher E, Suddason T. The PHD motif of Map3k1 activates cytokine-dependent MAPK signaling. Molecular & cellular oncology 2015; 2:e980659.

On 2014 Oct 30, jizhao li commented:

说的太好了……