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W. LaJean Chaffin, Ph.D.Dept. Immunology & Molecular Microbiology

Texas Tech University Health Sciences Center

Truth in pictures/images Image alteration outside science

Selection of examples Photojournalism

Image alteration in biomedical science Images are data Prevalence of inappropriate image alteration Prevention of publication of such images Detection inappropriate alteration What to do if you suspect problematic image Best practices or acceptable image manipulation

photoshop/photoshoppedtransitive verb: to alter (a digital image) with Photoshop software or other image-editing software especially in a way that distorts reality (as for deliberately deceptive purposes)

Photoshop registered trademark of Adobe Systems, Inc. for photo editing software

http://www.merriam-webster.com/dictionary/photoshop; access June 10, 2016

John C. Calhoun

Abraham Lincoln by Mathew Brady 1864

Head of Lincoln, flipped and body and background from Calhoun

Composite attributed to John Hicks may be the first “photoshopped”. After 1865 Lincoln assassination.

Stalin routinely airbrushed enemies or those out of favor out of photo

Hitler removed Goebbels when out of favor

Released Nov 5, 2008 by North Korean Central News Agency released 2 photos when Kim Jong-il thought to be very sick; image questioned by BBC and U.K. Times

King George VI and Queen Elizabeth with Canadian Prime Minster W.L. M. King, 1939

Images Stalin, Hitler, and King: Ye Olde Photoshoppe: The first ever altered images (including two pictures stitched together to make iconic portrait of Abraham Lincoln). Daily Mail, Feb 28, 2012 http://www.dailymail.co.uk/news/article-2107109/Iconic-Abraham-Lincoln-portrait-revealed-TWO-pictures-stitched-together.html

Airbrushing publicity shots: models, actors, public personalities

Generally accepted that publicity shots of these professionals may be airbrushed

Software packages to airbrush faces, change makeup, hair color, bodies e.g. PortraitPro15

2008 publicity shot Taylor Swift for album FearlessTaylor Swift: in pictures. Telegraph UK 16 Oct 2015. http://www.telegraph.co.uk/music/artists/taylor-swift-in-pictures/

Photojournalists 2003 Brian Walski of Los Angeles Times fired after

submitting a photograph of war in Iraq that composite of two images that was shared with other Tribune properties; “ was totally unacceptable and he violated our trust with our readers…”

2006 Rueters free lance photojournalist Beirut Adnan Hajj in Beirut enhanced photographs (e.g. added smoke, add missiles

2007 Toledo Blade fired award winning photojournalist Allan Detrich. Investigation showed that submitted 947 photographs January -March of which 79 digitally altered and 27 of altered photographs published in the newspaper and another 31 only online.

Images http://www.toledoblade.com/assets/pdf/TO16827415.PDF

2012 Award winning photojournalist Bryan Patrick fired by Sacramento Bee with printed apology to readers explaining background. After tip by reader lead to investigation image manipulation

2014 AP severed ties with Pulitzer Prize winning photojournalist Narcisco Contreras over altered photograph

Consequences of pointing out doctored images2012 photographer Johann Hattingh for The Citizen (a Johannesburg, South Africa) tweeted about a image on the cover was manipulated to remove bodies of two South Africans killed in blast in Kabul. He was fired.

IMAGES ARE DATA

Federal definition research misconduct NIH

Definition of Research MisconductResearch misconduct means fabrication, falsification, or plagiarism in proposing, performing, or reviewing research, or in reporting research results.(a) Fabrication is making up data or results and recording or reporting them.

(b) Falsification is manipulating research materials, equipment, or processes, or changing or omitting data or results such that the research is not accurately represented in the research record.

(c) Plagiarism is the appropriation of another person's ideas, processes, results, or words without giving appropriate credit.

(d) Research misconduct does not include honest error or differences of opinion. Last Updated on Mon, 2011-04-25 12:05.

http://ori.dhhs.gov/definition-misconduct accessed June 6, 2016 NSF similar http://www.nsf.gov/bfa/dias/policy/si/sipolicy.pdf;

http://www.nsf.gov/pubs/manuals/gpm05_131/gpm9.jsp#930

Office Research Integrity (DHHS) findings of misconduct involving images and/or mislabeling figures

2011: 7 of 13 cases (54%) 2012: 6 of 13 cases (46%) 2013: 1 of 5 cases (20%) 2014: 4 of 13 cases (31%) 2015: 3 of 14 cases (21%) 2016 to date: 3 of 5 cases (60%) (1 case -

41 published images, 1 case - over 100 Western blot images in record)

Journals and image manipulation policies stated

PLoS One. 2012. 7(12):e51928. doi: 10.1371/journal.pone.0051928. Epub 2012 Dec 19.Misconduct policies in high-impact biomedical journals.Bosch X, Hernández C, Pericas JM, Doti P, Marušić A.

Survey misconduct policies 399 high impact journals in 27 biomedical categories

140 (35.1%) explicit definitions of misconduct 154 (38.6%) image manipulation 112 (28.1%) use plagiarism checking services

Misconduct accounts for the majority of retracted scientific publications, Fang, FC, RG Steen, A Casedevall, 2012, PNAS 109; 17028

2047 PubMed articles on May 3, 2012 listed as retracted-21.3% error; 67.4% misconduct (fraud or suspected fraud 43.4%, duplicate publication 14.2%, plagiarism 9.5%). Percentage retracted because of fraud increase 10X since 1975

Prepublication screening J Cell Biology-2002

25% image problems 1% acceptance revoked for image

manipulation From Liz Williams executive Editor in Research integrity: Cell-induced stress, News

feature by David Cyranoski,Nature July 2014, 511. http://www.nature.com/news/research-integrity-cell-induced-stress-1.15507.

From Mike Rossner, Editor in Chief quoted. Image manipulation. Author of popular blog that charged fraud unmasked. By J Couzin-Frankel in Science. 2013 Jan 11;339(6116):132. doi: 10.1126/science.339.6116.132

EMBO Journals Experience-2011Level Attributes Action %

2014

I Cosmetic aberrations or mistakes with supporting source data and satisfactory author explanation

Allow revision, no report to institution

12

II Data “beautification” and undeclared manipulation that changes conclusions; available source data or new data

May allow revision, may report to institution

8

III Image manipulation with digital obfuscation (splicing, cloning, insertion, and selective deletion); no explanation; no source data

Reject and report < 0.5

Total 20.4

Pulverer, B, When things go wrong: correcting the scientrific record. 2015. The EMBO J 34:2483. (Editor in Chief)

EMBO journals Jana Christopher dedicated image analysis staff @20% accepted papers problem

Discuss with scientific editor (sometimes Chief Editor) May go back to authors and ask raw data for a series of

figures and may hint should insure carefully checked. Good sign if authors pick up problem

1% revoked for image manipulation If EMBO paper flagged on PubPeer will check immediately

but usually older papers and only low resolution. Interview for The image detective who roots out manuscript flaws by

Richard Van Noorden, June 2015, Nature in News: Q&A; http://www.nature.com/news/the-image-detective-who-roots-out-manuscript-flaws-1.17749

Other Journals June 2015 After interview with Jana Christopher, van

Noorden checked other journals UK Publisher The Company of Biologists

check their 5 journals; Development, J Cell Sci, J Exp Biol; Dis Models Mech; Biol Open

His own Nature Publishing Group-spot check and maybe key Western blots (2/wk). Spokesperson added not the resources to do all but continually review procedures

The prevalence of Inappropriate Image Duplication in Biomedical Research PublicationsBik, E.M., A. Casadevall, F.C. Fang; 2016 doi:10.112/mBio. 00809-16 (7 June 2016)

Screened 20,621 papers from 40 journals 1995-2014 (several publishers, different impact factors, identified in each journal with search term “western blot”

Examined all images (not just Western blot) on screen

Cat 1: simple duplication fig with >2 identical panels in same fig or different figs same paper-most common example reuse of ß-actin loading controls Panel A & B published

correction Could be honest

assembly error

Cat II: Duplication and repositioning; microscopic or blot images with clear regions of overlap, where one image had been shifted, rotated or reversed with respect to the other Panel A & B published

correction

Cat III. Duplication with alterations: images altered -complete or partial duplication of lanes, bands, or groups of cells. May have reversal with respect to each other, within the same image panel or between panels or figures. Includes “stamping”-defined area duplicated multiple times in same image; “patching”-part of image obscured by area of different bkgd, FACS images sharing conserved regions and other regions with data pts added or removed Panel A retracted Panel B corrected

Summary Bik et al findings 1 in 25 papers altered images (3.8%)

Relatively low 1995-2002 @2% and then 4%

Int J Oncol 12% to J Cell Bio 0.3% which screened images since 2002

Country of origin (PLOS ONE) 2013-2014: predicted probability India 1.93, China 1.89, Taiwan 1.2, US 0.6, UK 0.47, Japan 0.26, Germany 0.24

Summary Bik et al findings

Selected subset of 559 papers and looked other papers of first and last author 2,425 papers Found 269 papers problematic images. 11.1% which

greater original dataset: so increased probability if problematic image in one paper also another

Fig 4

Pisconti, et al (2006) J Cell Biol 172:233-244. Retracted for image manipulation Jan 22, 2013

Ricky Malhotra et al. J. Biol. Chem. 2010;285:13748-13760

©2010 by American Society for Biochemistry and Molecular Biology

Fig 1 Fig 2 Fig 4

May 24 & June 1 ORI reported findings of misconduct for Ricky Malhotra and Karen M. D’Souza respectively. Among findings for Malhotra relabeling of images in notebook and some used grant applications, presentation and one publication. Below images from a JBC paper attributed to Malhotra also in Fig 7 of this paper D’Souza for fabricated/falsified data in Western blot.

Detection inappropriate image manipulation or forensics

Example of discussion from Dept Computer Sciences- contains examples, refs http://www.cs.dartmouth.edu/farid/downloads/tutorials/digitalimageforensics.pdfSome subjects discussedFormat based, e.g. JPEGCameraPixelStatisticalGeometricPhysics

Image manipulation detection Image manipulation detection software

ORI has tools work with Photoshop https://ori.hhs.gov/forensic-tools for download https://ori.hhs.gov/advanced-forensic-actions (updated Oct 2015) and examples https://ori.hhs.gov/samples EMBO journals use ORI software JCB or Company of Biologists?

Image manipulation detection Image Data Integrity-consultants, Pres.

Mike Rossner who started screening at JCBhttp://www.imagedataintegrity.com/

Fourandsix founded by Kevin Connor and Hany Rafid Isitru (trust rating) free for consumers Tips on image forensic techniques Responsible use of imaging toolshttp://www.fourandsix.com/

http://www.imageforensic.org/ upload photo for analysis

Observation by readers Observation

If you suspect inappropriate image manipulation PubMed PubPeer Contact journal/editor Contact author Contact institution

If you suspect image manipulation

PubPeer- Post publication peer review (PPPR) Anonymous, if desired. Anonymity upheld in

state court with small caveat still under consideration https://pubpeer.com/

Can also post image on Imgurhttp://imgur.com/

Search for paper of interest or can select Recent to see which papers generating comments

Options PubMed

Pro Venue for posting comments

Con Unnecessarily embarrass authors-no mistake,

honest error Not anonymous-

Risk of lawsuit by authors Negative response from colleagues

Options- PubPeer-PPPR Pro

Anonymous Not all countries have strong or institutionalized policies on misconduct Some journals check

Con Unnecessarily embarrass authors honest error Lead to vigilantism, witch hunt May not remain anonymous. Lawsuit by Fazlul Sarkar to identify

anonymous commenters on PubPeer to sue them for defamation because contends comments cost job offer; ruling that must disclose 1 of 3 commenter IP address. Appeal by ACLU and Harold Varmus and Bruce Alberts filed support briefs; Wayne State revealed IP address of this commenter without objection around Jan 20, 2016 (appears against federal regulation, that complainant(s) kept confidential; free speech)

Anonymous whistle blowers-Committee on Publication Ethics Discussion Document

100s or 1000s requests to editors on plagiarism, image manipulation

COPE –generally editor take seriously if specific detailed evidence to support.

Pro- Potential to remain anonymous Journal tools to investigate Does not publicize authors Journal may notify institution if find issues

Con Not all editors or journals follow up

http://publicationethics.org/files/Whistleblowers_document_Final.pdf Jan 2013

Author Pro

Give chance to correct honest error on own initiative or respond that no issue

Might be able to contact anonymously

Con If misconduct, give chance to destroy records May cause friction between individuals

Institution Pro

USA:Procedures and policies for allegations Supposed to keep complainant

anonymous and investigate Report to funding agency

Con Institution may be reluctant to engage in

investigation Have seen that Wayne State did not fully

keep complainant anonymity

Recommendation from Retraction Watch co-founders Ivan Oransky and Adam Marcus re reporting suspected misconduct

Institutional Research Integrity Officers or equivalent May consider copying Office Research

Integrity or equivalent (investigation at institutional level but create paper trail)

Try PubPeerMarcus A and I Oransky. A Retraction: we gave bad advice. Lab Times 2015 Issue 6 p 37, Nov 25; http://www.labtimes-archiv.de/epaper/LT_15_06/#37/zPosted on Retraction Watch Nov 30, 2015 http://retractionwatch.com/2015/11/30/a-retraction-watch-retraction-our-2013-advice-on-reporting-misconduct-turns-out-to-have-been-wrong/

Suggestions to maintain integrity (sources last slide) Maintain an original copy of the image

in a medium that can not be altered, e.g. CD-R

Clearly label (and relate to experiment details)

NEVER work on or manipulate the original image (even generally accepted manipulations)

Generally acceptable manipulations

Applied to WHOLE image generally acceptable while applied to only one area questionable Small adjustments to brightness and contrast generally

acceptable BUT large changes may be questionable and truncate intensity information. Gamma adjustment to midtones usually Ok but need declaration.

Contrast or histogram stretch usually OK with low contrast images if no outliers

Small changes in color may be acceptable if color is not part of the information carried by the image but if part of data, e.g. colocalization, color change/distortion of dyes inappropriate

Generally acceptable manipulations

Cropping an image to remove true background generally acceptable BUT Remove part of image not support hypothesis is NOT Cropping may change context. Someone else may look at image

and see something that generates new idea or interpretation Cropping too much may leave too few pixels for reproduction in

journal/publication

Combining images is acceptable most journals ONLY if it is clear that two images involved and delineated, e.g. small gap, dark line. Depending on the experiment may be better to rerun an experiment on the same gel.

Cloning or copying objects from one part of the image or a different image is VERY questionable.

Additional suggestions Commercial image manipulation software.

Usually not recommended for biological images. Magnification. Since ultimate magnification is

often unknown, using magnification bars on the initial image that can be carried through is the best way to maintain scale.

Intensity measurements. Perform on raw data and data calibrated to a known standard.

Comparison. Images to be compared should be acquired under same conditions

Additional suggestions Size. Changing the size of image can introduce

artefacts. If the downsizing is not by a power of two the program has to “guess” while the power of two is a form of averaging which is somewhat better but still is a loss of resolution. Increasing the size also requires the software to create new pixels between the original ones and again the software has to “guess”. If the size must be changed it should only be once and the LAST manipulation of the image. If using Adobe Photoshop under :”Image Size” make sure the that the “Resample Image” is NOT checked.

Additional suggestions Avoid JPEG files. JPEG files are compressed

files (lossy compression) that also change XY resolution and intensity value of any given pixel. Happens each save. If must be used should be last step.

Image acquisition. For publication and sharing, acknowledge or describe in Methods the image acquisition and processing manipulations as appropriate.

How can we improve the picture

Likely >25,000 papers/yr in biomedical science have problematic, false images

Something is wrong with this picture

More journals use image checking More journals investigate allegations Complete development of misconduct policies in

countries and institutions Require use of authentication service prior to

submission Reviewers may identify more problematic if

assess images before reading text

Sources Societies and journals have articulated positions on imaging ethics. Some examples of

opinions, statements and policies http://www.springerlink.com/content/00311qw26613m261/fulltext.pdf Microscopy Society of America; http://www.microscopy.org/resources/digital_imaging.cfm http://www.springerlink.com/content/00311qw26613m261/fulltext.pdf Nature: http://www.nature.com/nature/authors/submissions/images/index.html Journal Cell biology in Instructions to Authors: http://jcb.rupress.org/site/misc/ifora.xhtml

Couzin, J. 2006. Don’t Pretty up that Picture just yet. Science 314:1866-1868 http://www.sciencemag.org/cgi/reprint/314/5807/1866.pdf?maxtoshow=&HITS=10&hits=10&RESULTFORMAT=&andorexacttitleabs=and&andorexactfulltext=and&searchid=1&FIRSTINDEX=0&volume=314&firstpage=1866&resourcetype=HWCIT

Rossner, M. and Yamada. K. M. 2004. What’s in a picture? The temptation of image manipulation J Cell Biol 166:11-15. http://jcb.rupress.org/cgi/reprint/166/1/11

The Office of Research Integrity has posted guidelines, questionable practices, case study and misconduct cases. http://ori.hhs.gov/education/products/RIandImages/default.html

Example from PubPeerBik and anonymoushttps://pubpeer.com/publications/F30641CE24023236C0C54A9229F69ESci. Rep liverColon: Frontiers in Pharmacology expression of concern investigate, higher education ministry in MalaysiaPeerJ expression of Concern; 4th in Recent patents on anti-cancer drugs