Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
Review
. 2018 Aug 17;113(5):39.
doi: 10.1007/s00395-018-0696-8.

Practical guidelines for rigor and reproducibility in preclinical and clinical studies on cardioprotection

Hans Erik Bøtker  1 , Derek Hausenloy  2   3   4   5   6 , Ioanna Andreadou  7 , Salvatore Antonucci  8 , Kerstin Boengler  9 , Sean M Davidson  2 , Soni Deshwal  8 , Yvan Devaux  10 , Fabio Di Lisa  8 , Moises Di Sante  8 , Panagiotis Efentakis  7 , Saveria Femminò  11 , David García-Dorado  12 , Zoltán Giricz  13   14 , Borja Ibanez  15 , Efstathios Iliodromitis  16 , Nina Kaludercic  8 , Petra Kleinbongard  17 , Markus Neuhäuser  18   19 , Michel Ovize  20   21 , Pasquale Pagliaro  11 , Michael Rahbek-Schmidt  22 , Marisol Ruiz-Meana  12 , Klaus-Dieter Schlüter  9 , Rainer Schulz  9 , Andreas Skyschally  17 , Catherine Wilder  2 , Derek M Yellon  2 , Peter Ferdinandy  13   14 , Gerd Heusch  23
Affiliations
Review

Practical guidelines for rigor and reproducibility in preclinical and clinical studies on cardioprotection

Hans Erik Bøtker et al. Basic Res Cardiol. .
No abstract available

PubMed Disclaimer

Conflict of interest statement

HEB and MRS are shareholders in CellAegis Inc. PF is founder and CEO of Pharmahungary, a Group of R&D companies.

Figures

Fig. 1
Fig. 1
Virtually created data on infarct size as percent of area at risk. For two groups, placebo vs. verum, data are presented as minimum and maximum (crosses), interquartile range from 25 to 75% (box), mean (square), and median (line) in a box plot. The maximum value of the verum group is identified as outlier. All single values (squares) are depicted. P value as the result of a two-sided Student t test
Fig. 2
Fig. 2
Virtually created data on cellular viability. Sequential data are analyzed throughout the time course (baseline and after incubation) of the different experimental groups (time control and hypoxia/reoxygenation (H/R) with buffer, placebo, and verum, respectively). Data are presented as mean ± standard deviation. P values as the result of two-way repeated-measures ANOVA (group, time) with subsequent post hoc tests are reported
Fig. 3
Fig. 3
Virtually created data on left ventricular developed pressure during ischemia (Isch) and reperfusion (Rep). For two groups, placebo vs. verum, data are presented as mean ± standard deviation. P values as the result of two-way repeated-measures ANOVA (group, time) with subsequent post hoc tests are reported
Fig. 4
Fig. 4
Representative figure of a good cell preparation (left) and a bad preparation (right). Please note the difference in the number of rod-shaped cells
Fig. 5
Fig. 5
a Calcium transient recording in a fluo-4 loaded rat cardiomyocyte submitted to field stimulation (1 Hz, acquisition set at 40 images/s). After a period of stabilization, the cardiomyocyte is subjected to a single pulse of 10 mmol/L caffeine (red arrow) to induce maximal SR calcium release. The sequence of images shows diastolic and systolic calcium levels during pacing and after caffeine pulse. b Mitochondrial Ca2+ uptake throughout time in response to sarcoplasmic reticulum Ca2+ release (induced by a single pulse of 10 mmol/L caffeine, red arrow) in digitonin-permeabilized rhod-2 loaded mouse cardiomyocytes, under control conditions (black circles) or in the presence of 10 µmol/L of the mitochondrial calcium uniporter blocker Ru360 (white circles). Data correspond to mean ± standard error of the mean of n = 8 cardiomyocytes per group. Images correspond to a sarcolemmal-permeabilized cardiomyocyte loaded with rhod-2, before and after caffeine addition
Fig. 6
Fig. 6
a Sequence of a calcein-loaded mouse cardiomyocyte with rigor contracture during the first 5 min of reperfusion (60 × magnification). Because calcein is entrapped in mitochondria, the decay of green fluorescence reflects mitochondrial permeabilization. b Quantification of total mitochondrial calcein release at 15 min reperfusion, with respect to maximal mitochondrial calcein release achieved after the addition of atractyloside (ATR), in isolated cardiomyocytes from wild-type (WT) and cyclophilin-D knock-out (CyD-KO) mouse hearts, previously submitted to either 15 or 25 min ischemia. The inset shows mitochondrial calcein kinetics [changes in fluorescence with respect to the initial value, F/F0)] throughout reperfusion, indicating mitochondrial permeability transition pore (mPTP) opening in the different groups of cells. Maximal mitochondrial calcein release was induced in WT cardiomyocytes treated with 20 µmol/L atractyloside. Data are expressed as mean ± SEM of 5–6 cardiomyocytes per group. Modified from [358]
Fig. 7
Fig. 7
Diagram of a gravitational Langendorff perfused heart rig with an isolated rat heart
Fig. 8
Fig. 8
In the Langendorff configuration, buffer from the aortic cannula is prevented by the aortic valve from entering the heart, and passes instead through the coronary arteries (not shown), which originate from the base of the aorta. In the working heart configuration, buffer enters the left atrium via the atrial cannula, passes through the mitral valve to the left ventricle (LV), then is ejected during systole through the aortic valve into the aortic cannula
Fig. 9
Fig. 9
Original ECG recorded in one pig undergoing remote ischemic perconditioning (RPER) and one undergoing placebo intervention (PLA). ST-segment elevation is displayed as amplitude difference between two points (vertical red lines) 30 ms before the P wave and 20 ms after the J-point, respectively [252]
Fig. 10
Fig. 10
a Atrial trabeculae on the surface of the appendage being dissected. b Atrial trabecula mounted between two platinum pacing electrodes. c Functional readout from the atrial trabecula showing the change in contractile function (difference in tension) during hypoxia and reoxygenation. d The recovery of contractile function at the end of the experiment is greater in an atrial trabecula that had been preconditioned prior to hypoxia and reoxygenation
Fig. 11
Fig. 11
Different omics approaches from the genome to metabolome. Current technologies so far allow unbiased analysis of all genes and transcripts. Based on transcriptomic fingerprints, in silico prediction of molecular targets can be achieved. Predicted targets must always be validated at the protein level. Other omics technologies are rapidly emerging; however, their use is currently limited to assess only fractions of the total epigenetic/transcriptomic, proteomic and metabolomic pool
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. (CHMP) CfMPfHU (2010) Guideline on missing data in confirmatory clinical trials. http://www.ema.europa.eu/docs/en_GB/document_library/Scientific_guidelin.... Accessed 6 Aug 2018
    1. Abas L, Bogoyevitch MA, Guppy M. Mitochondrial ATP production is necessary for activation of the extracellular-signal-regulated kinases during ischaemia/reperfusion in rat myocyte-derived H9c2 cells. Biochem J. 2000;349:119–126. doi: 10.1042/bj3490119. - DOI - PMC - PubMed
    1. Abogresha NM, Greish SM, Abdelaziz EZ, Khalil WF. Remote effect of kidney ischemia-reperfusion injury on pancreas: role of oxidative stress and mitochondrial apoptosis. Arch Med Sci. 2016;12:252–262. doi: 10.5114/aoms.2015.48130. - DOI - PMC - PubMed
    1. Ackers-Johnson M, Li PY, Holmes AP, O’Brien SM, Pavlovic D, Foo RS. A simplified, Langendorff-free method for concomitant isolation of viable cardiac myocytes and nonmyocytes from the adult mouse heart. Circ Res. 2016;119:909–920. doi: 10.1161/CIRCRESAHA.116.309202. - DOI - PMC - PubMed
    1. Adameova A, Goncalvesova E, Szobi A, Dhalla NS. Necroptotic cell death in failing heart: relevance and proposed mechanisms. Heart Fail Rev. 2016;21:213–221. doi: 10.1007/s10741-016-9537-8. - DOI - PubMed

MeSH terms

Substances