​Karakashevska and colleagues (2024) aim to examine the extraretinal representation of visual symmetry presented in a virtual reality environment. Previous research had demonstrated that individuals can detect symmetry when the symmetry is represented on a perspective plane, slanted away from the viewer. In electroencephalography (EEG), perceived symmetry is marked by an Event Related Potential (ERP) called a Sustained Posterior Negativity (SPN). When symmetry is presented on a perspective plane in comparison to front-on (frontoparallel), the SPN is reduced, termed the perspective cost. Here, Karakashevska et al., (2024) will determine if presenting symmetry on a perspective plane in a virtual reality (VR) environment will reduce the perspective cost with the addition of 3D depth cues. Specifically, participants will be requested to detect symmetry or luminance of a stimulus presented in a VR environment whilst wearing an EEG. The authors hypothesize that no perspective cost will be identified between symmetry presented on a frontoparallel plane versus symmetry on a perspective plane. Furthermore, the authors will examine the impact of task within the virtual environment on symmetry processing. They hypothesize that a task focused on the regularity of the stimuli will result in a larger amplitude of the SPN than a luminance task. This design enables the authors to pinpoint immersive environments as providing cues critical in overcoming perspective cost.

 

The Stage 1 manuscript was evaluated by two expert reviewers across three rounds. Following in-depth review and responses from the authors, the recommender has determined that Stage 1 criteria was met and has awarded in-principle acceptance (IPA).

 

URL to the preregistered Stage 1 protocol: https://osf.io/7pnxu
 
Level of bias control achieved: Level 6. No part of the data or evidence that will be used to answer the research question yet exists and no part will be generated until after IPA. 
 
List of eligible PCI RR-friendly journals:

 

 

Karakashevska, E., Batterley, M. & Makin, A. D. J. (2024). They look virtually the same: extraretinal representation of symmetry in virtual reality. In principle acceptance of Version 3 by Peer Community in Registered Reports. https://osf.io/7pnxu

The Spatial-Numerical Association of Response Codes (SNARC) effect refers to the fact that smaller numbers receive faster responses with the left hand, and larger numbers with the right hand (Dehaene et al., 1993). This robust finding implies that numbers are associated with space, being represented on a mental number line that progresses from left to right. The SNARC effect is held to depend on relative number magnitude, with the mental number line dynamically adjusting to the numerical range used in a given context. This characterisation is based on significant effects of relative number magnitude, with no significant influence of absolute number magnitude. However, a failure to reject the null hypothesis is not firm evidence for the absence of an effect. In this Registered Report, Roth and colleagues (2024) report two large-sample online experiments, with a Bayesian statistical approach to confirm—or refute—a role for absolute number magnitude in modulating the classic SNARC effect (smallest effect size of interest, d = 0.15).

 

Experiment 1 closely followed Dehaene’s (1993) original methods, and found strong evidence for an influence of relative magnitude, and moderate-to-strong evidence against an influence of absolute magnitude. Experiment 2 was designed to exclude some potential confounds in the original method, and this second experiment found strong evidence for both relative and absolute magnitude effects, of comparable effect sizes (in the range of d = .24 to .42). This registered study demonstrates that the SNARC effect is not ‘fully flexible’, in the sense of depending only on relative number magnitude; it is also shaped by absolute magnitude.

 

This Stage 2 manuscript was evaluated by the recommender and one external reviewer. Following appropriate minor revisions, the recommender judged that the manuscript met the Stage 2 criteria for recommendation.

 

URL to the preregistered Stage 1 protocol: https://osf.io/ae2c8
 
Level of bias control achieved: Level 6. No part of the data or evidence that was used to answer the research question was generated until after IPA.
 
List of eligible PCI RR-friendly journals:
 

• Advances in Cognitive Psychology
• Collabra: Psychology
• Experimental Psychology
• Journal of Cognition
• Peer Community Journal
• PeerJ
• Psychology of Consciousness: Theory, Research, and Practice
• Royal Society Open Science
• Studia Psychologica
• Swiss Psychology Open

 
References

 

1. Dehaene, S., Bossini, S., & Giraux, P. (1993). The mental representation of parity and number magnitude. Journal of Experimental Psychology: General, 122, 371–396. https://doi.org/10.1037/0096-3445.122.3.371

 

2. Roth, L., Caffier, J., Reips, U.-D., Nuerk, H.-C., Overlander, A. T. & Cipora, K. (2023). One and only SNARC? Spatial-Numerical Associations are not fully flexible and depend on both relative and absolute magnitude [Stage 2]. Acceptance of Version 3 by Peer Community in Registered Reports. https://osf.io/epnd4

A recent study has reported that the extent of cerebral small vessel disease (cSVD) shows associations with dynamic functional connectivity measures obtained from resting state functional MRI scans (Schlemm et al, 2022). Specifically, when the functional scan was parsed into time spent in discrete brain states, the proportion of time spent in the two most-occupied states was negatively related to a structural indicator of cSVD (volume of white matter hyperintensities of presumed vascular origin). This measure of 'fractional occupancy' was also associated with cognitive impairment as indicated by longer time to complete part B of the Trail Making Test. These findings were based on the analysis of data from 988 participants in the Hamburg City Health Study (HCHS).

 

In the present Registered Report, Schlemm (2023) will test whether these associations can be replicated in an independent sample of participants from the HCHS, not included in the earlier analysis (projected N for new analysis ~1500). In addition to the two main hypothesis tests, an exploratory multiverse analysis will be reported, systematically varying some key parameters of the MRI processing pipeline to provide further information about the robustness of the outcome of the primary hypothesis test. 

 

The Stage 1 plan was refined over two rounds of review by two relevant experts, with additional input from the recommender on the specification of the registered plan. Both reviewers are satisfied that the plan constitutes an appropriate approach to this question, and on the basis of their comments and his own evaluation, the recommender judged that the Stage 1 report meets the criteria for in-principle acceptance.

 

URL to the preregistered Stage 1 protocol: https://osf.io/9yhzc
 
Level of bias control achieved: Level 2. At least some data/evidence that will be used to answer the research question has been accessed and partially observed by the authors, but the authors certify that they have not yet observed the key variables within the data that will be used to answer the research question AND they have taken additional steps to maximise bias control and rigour.
 
List of eligible PCI RR-friendly journals:

 

 

 

1. Schlemm, E., Frey, B. M., Mayer, C., Petersen, M., Fiehler, J., Hanning, U., Kühn, S., Twerenbold, R., Gallinat, J., Gerloff, C., Thomalla, G. & Cheng, B. (2022). Equalization of brain state occupancy accompanies cognitive impairment in cerebral small vessel disease. Biological Psychiatry, 92, 592-602. https://doi.org/10.1016/j.biopsych.2022.03.019

 

2. Schlemm, E. (2023). Functional MRI brain state occupancy in the presence of cerebral small vessel disease – pre-registration for a replication analysis of the Hamburg City Health Study. In principle acceptance of Version 1.5 by Peer Community in Registered Reports. https://osf.io/9yhzc

In a previous analysis of data from 988 participants in the Hamburg City Health Study (HCHS), Schlemm and colleagues (2022) reported significant associations between the extent of cerebral small vessel disease (cSVD) and dynamic functional connectivity measures from resting state fMRI. Specifically, the volume of white matter hyperintensities of presumed vascular origin, a structural indicator of cSVD, was negatively related to the proportion of time (‘fractional occupancy’) spent in the two most occupied functional brain states. Reduced fractional occupancy was also associated with longer times to complete part B of the Trail Making Test.

 

In the present Registered Report, Ingwersen and colleagues (2023) successfully replicated these associations between structural, functional and cognitive measures in a sample of 1651 HCHS participants not included in the earlier study. An exploratory multiverse analysis found that the associations were generally robust to different brain parcellation and confound regression strategies. These replicable patterns reinforce the idea that cSVD may disrupt the brain’s ability to enter and maintain distinct functional modes, and that these changes in functional dynamics are predictive of cognitive impairment.

 

The Stage 2 manuscript was assessed over one round of in-depth review. The recommender judged that responses to reviewer comments were appropriate, and that the manuscript met the Stage 2 criteria for recommendation.

 

URL to the preregistered Stage 1 protocol: https://osf.io/9yhzc
 
Level of bias control achieved: Level 2. At least some data/evidence that was used to answer the research question had been accessed and partially observed by the authors prior to Stage 1 in-principle acceptance, but the authors certify that they had not yet observed the key variables within the data that were used to answer the research question AND they took additional steps to maximise bias control and rigour.
 
List of eligible PCI RR-friendly journals:
 

 

 

1. Schlemm, E., Frey, B. M., Mayer, C., Petersen, M., Fiehler, J., Hanning, U., Kühn, S., Twerenbold, R., Gallinat, J., Gerloff, C., Thomalla, G. & Cheng, B. (2022). Equalization of brain state occupancy accompanies cognitive impairment in cerebral small vessel disease. Biological Psychiatry, 92, 592-602. https://doi.org/10.1016/j.biopsych.2022.03.019

 

2. Ingwersen, T., Mayer, C., Petersen, M., Frey, B. M., Fiehler, J., Hanning, U., Kühn, S., Gallinat, J., Twerenbold, R., Gerloff, C., Cheng, B., Thomalla, G. & Schlemm, E. (2023). Functional MRI brain state occupancy in the presence of cerebral small vessel disease -- a pre-registered replication analysis of the Hamburg City Health Study. Acceptance of Version 2.01 by Peer Community in Registered Reports. https://github.com/csi-hamburg/HCHS-brain-states-RR/blob/f9d00adbbcf9593d8d191bf5b93912141b80ab1b/manuscript/build/main.pdf

 

 

Over the past decade there has been an exponential increase in the number of research papers utlising nanoparticles for biological applications such as intracellular sensing [1, 2], theranostics [3-5] and more recently drug delivery and precision medicine [6, 7]. Despite the success stories, there is a disconnect regarding current dogma on issues such as nanoparticle uptake and trafficking, nanoparticle delivery via the enhanced permeability and retention (EPR) effect, and endosomal escape. Critical re-evaluation of these concepts both conceptually and experimentally is needed for continued advancement in the field.

 

For this preregistration, Said et al. (2024) focus on nanoparticle intracellular trafficking, specifically endosomal escape [8]. The current consensus in the literature is that nanoparticles enter cells via endocytosis [9, 10] but reportedly just 1-2% of nanoparticles/ nanoparticle probes escape endosomes and enter the cytoplasm [11-13]. There is therefore an apparent paradox over how sensing nanoparticles can detect their intended targets in the cytoplasm if they are trapped within the cell endosomes. To address this fundamental issue of nanoparticle endosomal escape, Lévy and coworkers are carrying out replication studies to thoroughly and transparently replicate the most influential papers in the field of nanoparticle sensing. The aim of these replication studies is twofold: to establish a robust methodology to study endosomal escape of nanoparticles, and to encourage discussions, transparency and a step-change in the field.  

 

 

For this replication study, the authors classified papers on the topic of nanoparticle sensing and subsequently ranked them by number of citations.  Based on this evaluation they selected a paper by Zhu and colleagues [14] entitled “Carbon-Dot-Based Dual-Emission Nanohybrid Produces a Ratiometric Fluorescent Sensor for In Vivo Imaging of Cellular Copper Ions” for their seminal replication study.  To determine the reproducibility of the results from Zhu et al., the authors aim to establish the proportion of endosomal escape of the nanoparticles, and to examine the data in a biological context relevant to the application of the probe.

 

 

The authors plan to replicate the exact conditions reported in the materials and methods section of the selected paper such as nanoparticle probe synthesis of CdSe@C-TPEA nanoparticles, assessment of particle size, stability and reactivity and effect on cells (TEM, pH experiments, fluorescent responsivity to metal ions and cell viability). In addition, Said et al., plan to include further experimental characterisation to complement the existing study by Zhu and colleagues. They will incorporate additional controls and methodology to determine the intracellular location of nanoparticle probes in cells including: quantifying excess fluorescence in the culture medium, live cell imaging analysis, immunofluorescence with endosomal and lysosomal markers, and electron microscopy of cell sections. The authors will also include supplementary viability studies to assess the impact of the nanoparticles on HeLa cells as well as an additional biologically relevant cell line (for use in conjunction with the HeLa cells as per the original paper).

 

The Stage 1 manuscript underwent two rounds of thorough in-depth review. After considering the detailed responses to the reviewers' comments, the recommenders  determined that the manuscript met the Stage 1 criteria and awarded in-principle acceptance (IPA).  

 

The authors have thoughtfully considered their experimental approach to the replication study, whilst acknowledging any potential limitations. Given that conducting such a replication study is novel in the field of Nanotechnology and there is currently no ‘gold standard’ approach in doing so, the authors have showed thoughtful regard of statistical analysis and unbiased methodology where possible.

 

Based on current information, this study is the first use of preregistration via Peer Community in Registered Reports and the first formalised replication study in Nanotechnology for Biosciences. The outcomes of this of this study will be significant both scientifically and in the wider context in discussion of the scientific method.

 

 
Level of bias control achieved: Level 6. No part of the data or evidence that will be used to answer the research question yet exists and no part will be generated until after IPA.
 
List of eligible PCI RR-friendly Journals:

 

 

1. Howes, P. D., Chandrawati, R., & Stevens, M. M. (2014). Colloidal nanoparticles as advanced biological sensors. Science, 346(6205), 1247390. https://doi.org/10.1126/science.1247390

 

2. Liu, C. G., Han, Y. H., Kankala, R. K., Wang, S. B., & Chen, A. Z. (2020). Subcellular performance of nanoparticles in cancer therapy. International Journal of Nanomedicine, 675-704. https://doi.org/10.2147/IJN.S226186

 

3. Tang, W., Fan, W., Lau, J., Deng, L., Shen, Z., & Chen, X. (2019). Emerging blood–brain-barrier-crossing nanotechnology for brain cancer theranostics. Chemical Society Reviews, 48(11), 2967-3014. https://doi.org/10.1039/C8CS00805A

 

4. Yoon, Y. I., Pang, X., Jung, S., Zhang, G., Kong, M., Liu, G., & Chen, X. (2018). Smart gold nanoparticle-stabilized ultrasound microbubbles as cancer theranostics. Journal of Materials Chemistry B, 6(20), 3235-3239. https://doi.org/10.1039%2FC8TB00368H

 

5. Lin, H., Chen, Y., & Shi, J. (2018). Nanoparticle-triggered in situ catalytic chemical reactions for tumour-specific therapy. Chemical Society Reviews, 47(6), 1938-1958. https://doi.org/10.1039/C7CS00471K

 

6. Hou, X., Zaks, T., Langer, R., & Dong, Y. (2021). Lipid nanoparticles for mRNA delivery. Nature Reviews Materials, 6(12), 1078-1094. https://doi.org/10.1038/s41578-021-00358-0

 

7. Mitchell, M. J., Billingsley, M. M., Haley, R. M., Wechsler, M. E., Peppas, N. A., & Langer, R. (2021). Engineering precision nanoparticles for drug delivery. Nature Reviews Drug Discovery, 20(2), 101-124. https://doi.org/10.1038/s41573-020-0090-8

 

8. Said, M., Gharib, M., Zrig, S., & Lévy, R. (2024). Replication of “Carbon-Dot-Based Dual-Emission Nanohybrid Produces a Ratiometric Fluorescent Sensor for In Vivo Imaging of Cellular Copper Ions”. In principle acceptance of Version 3 by Peer Community in Registered Reports. https://osf.io/qbxpf

 

9. Behzadi, S., Serpooshan, V., Tao, W., Hamaly, M. A., Alkawareek, M. Y., Dreaden, E. C., ... & Mahmoudi, M. (2017). Cellular uptake of nanoparticles: Journey inside the cell. Chemical Society Reviews, 46(14), 4218-4244. https://doi.org/10.1039/C6CS00636A

 

10. de Almeida, M. S., Susnik, E., Drasler, B., Taladriz-Blanco, P., Petri-Fink, A., & Rothen-Rutishauser, B. (2021). Understanding nanoparticle endocytosis to improve targeting strategies in nanomedicine. Chemical society reviews, 50(9), 5397-5434. https://doi.org/10.1039/D0CS01127D

 

11. Smith, S. A., Selby, L. I., Johnston, A. P., & Such, G. K. (2018). The endosomal escape of nanoparticles: toward more efficient cellular delivery. Bioconjugate Chemistry, 30(2), 263-272. http://dx.doi.org/10.1021/acs.bioconjchem.8b00732

 

12. Cupic, K. I., Rennick, J. J., Johnston, A. P., & Such, G. K. (2019). Controlling endosomal escape using nanoparticle composition: current progress and future perspectives. Nanomedicine, 14(2), 215-223. https://doi.org/10.2217/nnm-2018-0326

 

13. Wang, Y., & Huang, L. (2013). A window onto siRNA delivery. Nature Biotechnology, 31(7), 611-612. https://doi.org/10.1038/nbt.2634

 

14. Zhu, A., Qu, Q., Shao, X., Kong, B., & Tian, Y. (2012). Carbon-dot-based dual-emission nanohybrid produces a ratiometric fluorescent sensor for in vivo imaging of cellular copper ions. Angewandte Chemie (International ed. in English), 51(29), 7185-7189. https://doi.org/10.1002/anie.201109089