Face Short-Term Memory-Related Electroencephalographic Patterns can Differentiate Multi- versus Single-Domain Amnestic Mild Cognitive Impairment.

Amnestic mild cognitive impairment (aMCI) is characterized by memory deficits alone (single-domain, sd-aMCI) or associated with other cognitive disabilities (multi-domain, md-aMCI). The present study assessed the patterns of electroencephalographic (EEG) activity during the encoding and retrieval phases of short-term memory in these two aMCI subtypes, to identify potential functional differences according to the neuropsychological profile. Continuous EEG was recorded in 43 aMCI patients, whose 16 sd-aMCI and 27 md-aMCI, and 36 age-matched controls (EC) during delayed match-to-sample tasks for face and letter stimuli. At encoding, attended stimuli elicited parietal alpha (8-12 Hz) power decrease (desynchronization), whereas distracting stimuli were associated with alpha power increase (synchronization) over right central sites. No difference was observed in parietal alpha desynchronization among the three groups. For attended faces, the alpha synchronization underlying suppression of distracting letters was reduced in both aMCI subgroups, but more severely in md-aMCI cases that differed significantly from EC. At retrieval, the early N250r recognition effect was significantly reduced for faces in md-aMCI as compared to both sd-aMCI and EC. The results suggest a differential alteration of working memory cerebral processes for faces in the two aMCI subtypes, face covert recognition processes being specifically altered in md-aMCI.

A unique role for Kv3 voltage-gated potassium channels in starburst amacrine cell signaling in mouse retina

Direction-selective retinal ganglion cells show an increased activity evoked by light stimuli moving in the preferred direction. This selectivity is governed by direction-selective inhibition from starburst amacrine cells occurring during stimulus movement in the opposite or null direction. To understand the intrinsic membrane properties of starburst cells responsible for direction-selective GABA release, we performed whole-cell recordings from starburst cells in mouse retina. Voltage-clamp recordings revealed prominent voltage-dependent K+ currents. The currents were mostly blocked by 1 mm TEA, activated rapidly at voltages more positive than -20 mV, and deactivated quickly, properties reminiscent of the currents carried by the Kv3 subfamily of K+ channels. Immunoblots confirmed the presence of Kv3.1 and Kv3.2 proteins in retina and immunohistochemistry revealed their expression in starburst cell somata and dendrites. The Kv3-like current in starburst cells was absent in Kv3.1-Kv3.2 knock-out mice. Current-clamp recordings showed that the fast activation of the Kv3 channels provides a voltage-dependent shunt that limits depolarization of the soma to potentials more positive than -20 mV. This provides a mechanism likely to contribute to the electrical isolation of individual starburst cell dendrites, a property thought essential for direction selectivity. This function of Kv3 channels differs from that in other neurons where they facilitate high-frequency repetitive firing. Moreover, we found a gradient in the intensity of Kv3.1b immunolabeling favoring proximal regions of starburst cells. We hypothesize that this Kv3 channel gradient contributes to the preference for centrifugal signal flow in dendrites underlying direction-selective GABA release from starburst amacrine cells.

Prognostic value of cortically induced motor evoked activity by TMS in chronic stroke: caveats from a very revealing single clinical case

Background: We report the case of a chronic stroke patient (62 months after injury) showing total absence of motor activity evoked by transcranial magnetic stimulation (TMS) of spared regions of the left motor cortex, but near-to-complete recovery of motor abilities in the affected hand. Case presentation: Multimodal investigations included detailed TMS based motor mapping, motor evoked potentials (MEP), and Cortical Silent period (CSP) as well as functional magnetic resonance imaging (fMRI) of motor activity, MRI based lesion analysis and Diffusion Tensor Imaging (DTI) Tractography of corticospinal tract (CST). Anatomical analysis revealed a left hemisphere subinsular lesion interrupting the descending left CST at the level of the internal capsule. The absence of MEPs after intense TMS pulses to the ipsilesional M1, and the reversible suppression of ongoing electromyographic (EMG) activity (indexed by CSP) demonstrate a weak modulation of subcortical systems by the ipsilesional left frontal cortex, but an inability to induce efficient descending volleys from those cortical locations to right hand and forearm muscles. Functional MRI recordings under grasping and finger tapping patterns involving the affected hand showed slight signs of subcortical recruitment, as compared to the unaffected hand and hemisphere, as well as the expected cortical activations. Conclusions: The potential sources of motor voluntary activity for the affected hand in absence of MEPs are discussed. We conclude that multimodal analysis may contribute to a more accurate prognosis of stroke patients.

An Ion Transport-Independent Role for the Cation-Chloride Cotransporter KCC2 in Dendritic Spinogenesis In Vivo.

The neuron-specific K-Cl cotransporter, KCC2, is highly expressed in the vicinity of excitatory synapses in pyramidal neurons, and recent in vitro data suggest that this protein plays a role in the development of dendritic spines. The in vivo relevance of these observations is, however, unknown. Using in utero electroporation combined with post hoc iontophoretic injection of Lucifer Yellow, we show that premature expression of KCC2 induces a highly significant and permanent increase in dendritic spine density of layer 2/3 pyramidal neurons in the somatosensory cortex. Whole-cell recordings revealed that this increased spine density is correlated with an enhanced spontaneous excitatory activity in KCC2-transfected neurons. Precocious expression of the N-terminal deleted form of KCC2, which lacks the chloride transporter function, also increased spine density. In contrast, no effect on spine density was observed following in utero electroporation of a point mutant of KCC2 (KCC2-C568A) where both the cotransporter function and the interaction with the cytoskeleton are disrupted. Transfection of the C-terminal domain of KCC2, a region involved in the interaction with the dendritic cytoskeleton, also increased spine density. Collectively, these results demonstrate a role for KCC2 in excitatory synaptogenesis in vivo through a mechanism that is independent of its ion transport function.

The JAK2/STAT3 pathway in the embryonic chick heart submitted to anoxia, reoxygenation and oxidant stress

SUMMARYAim: The embryonic/fetal heart is highly sensitive to oxygenation level and a transient uteroplacental hypoperfusion can lead to oxyradicals overproduction. Information about the molecular mechanisms underlying ischemia-reperfusion (I-R) injury in the developing heart is lacking. The Janus Kinase 2 / Signal Transducer and Activator of Transcription 3 (JAK2/STAT3) pathway, required for cardiogenesis and involved in protection of the adult heart against I-R, could also play a key role in the response of the fetal myocardium to transient oxygen deprivation. The aim of the study was to characterize the involvement of JAK2/STAT3 pathway and its interaction with other signalling pathways in the developing heart transiently submitted to anoxia. Furthermore, the response of the embryonic heart to an exogenous oxidant stress (H2O2) in comparison to reoxygenation-induced endogenous oxyradicals has been investigated.Methods: Hearts isolated from 4-day-old chick embryos were submitted to anoxia (30min) and reoxygenation (80min) with or without the antioxidant MPG, the JAK2/STAT3 inhibitor AG490 or exposed to H202 (50|iM-lmM). The time course of phosphorylation of STAT3atyr0Sine7 and Reperfusion Injury Salvage Kinase (RISK) proteins (PI3K, Akt, GSK3B, Glycogen Synthase and ERK2) was determined in homogenate" and in enriched nuclear and cytoplasmic fractions. The STAT3 DNA-binding was determined by EMSA and the expression of STAT3 specific target genes by RT-PCR. The chrono-, dromo- and inotropic disturbances were also investigated by ECG and mechanical recordings.Results: Phosphorylation of STATSaP (P-Tyr STAT3a) was increased by reoxygenation and reduced by MPG or AG490. STAT3 and GSK36 were detected both in nuclear and cytoplasmic fractions while PI3K, Akt, GS and ERK2 were restricted to cytoplasm. Reoxygenation led to nuclear accumulation of STAT3 but unexpectedly without DNA- binding. AG490 decreased the reoxygenation-induced phosphorylation of STABa^, Akt, GS and ERK2 and phosphorylation/inhibition of GSK3B in the nucleus, exclusively. Inhibition of JAK2/STAT3 delayed recovery of atrial rate, worsened RR. variability and prolonged arrhythmias compared to control hearts. Cardiac activity was altered only at concentrations >500μΜ of H2O2. Moreover, ImM of H2O2 suppressed atrial activity in 45% of the hearts, atrioventricular conduction in 66% and augmented P-Tyr STAT3awhich led to an increase in the DNA-binding but no change in the expression of three STAT3 specific target genes (iNOS, MnSOD, Cox-2).Conclusion: In the developing heart, besides its nuclear translocation without transcriptional activity, ROS-activated STAT3a can rapidly interact with RISK proteins present in nucleus and cytoplasm and reduce the anoxia-reoxygenation-induced arrhythmias. Moreover, the embryonic heart is highly resistant to H2O2 and the atrial region is the less affected. The role of JAK2/STAT3 in the response to reoxygenation-induced oxyradicals is different from the response to strong exogenous oxidant stress where STAT3 DNA-binding activity is increased. Such findings provide a first step in understanding the modulation of signalling cascades in the fetal heart submitted to transient intrauterine oxygen deprivation.RESUMEIntroduction: Le coeur embryonnaire et foetal est très sensible au manque d'oxygène et une hypoperfusion utéroplacentaire transitoire peut conduire à une surproduction d'espèces radicalaires (ROS). Dans le coeur en développement les mécanismes moléculaires impliqués en situation d'ischémie-reperfusion (I-R) ne sont pas connus. La voie de signalisation JAK2/STAT3 (Janus Kinase 2 / Signal Transducer and Activator of Transcription 3), impliquée aussi bien dans la cardiogenèse précoce que dans la protection du coeur adulte contre l'I-R, pourrait jouer un rôle clé dans la réponse du myocarde foetal à un déficit en oxygène. Cette étude a permis d'étudier le rôle de la voie JAK2/STAT3 et son interaction avec d'autres voies de signalisation dans un modèle de coeur embryonnaire soumis à un épisode anoxique. En outre, les effets du stress oxydant endogène provoqué par la réoxygénation ont été comparés à ceux du stress oxydatif exogène induit par du peroxyde d'hydrogène (H2O2).Méthodes: Des coeurs isolés d'embryons de poulet âgés de 4 jours ont été soumis à une anoxie (30min) suivie d'une réoxygénation (80min) en présence ou non de l'antioxydant MPG et de l'inhibiteur de JAK2/STAT3 AG490 ou exposés à de 1Ή202 (50μΜ-1πιΜ). L'évolution temporelle de la phosphorylation de 8ΤΑΤ3α*ΓΟδίη6705 (P-Tyr STAT3a) et celle de la phosphorylation des protéines de la voie RISK (Reperfusion Injury Salvage Kinase: PI3K, Akt, GSK3B, glycogène synthase GS et ERK2) ont été déterminés dans l'homogénat et dans les fractions nucléaire et cytopiasmique du myocarde. La liaison de STAT3 à l'ADN a été déterminée par EMSA et l'expression de gènes cibles de STAT3 (iNOS, MnSOD, Cox2) par RT-PCR. Les effets chrono-, dromo- et inotropes ont été déterminés par les enregistrements de l'ECG et de l'activité contractile ventriculaire.Résultats: STAT3 et GSK3B étaient présents dans les fractions nucléaire et cytopiasmique tandis que PI3K, Akt, GS et ERK2 n'étaient détectées que dans la fraction cytopiasmique. L'augmentation de P-Tyr STAT3a provoquée par la réoxygénation était significativement réduite par le MPG ou PAG490. La réoxygénation entraînait l'accumulation nucléaire de STAT3, mais étonnamment sans liaison avec l'ADN. A la réoxygénation TAG490 diminuait la phosphorylation d'Akt, GS et ERK2 ainsi que celle de GSK36 mais exclusivement dans la fraction nucléaire. L'inhibition de JAK2/STAT3 retardait également la récupération du rythme cardiaque et prolongeait la durée des arythmies. L'activité cardiaque n'était perturbée par de ΓΗ2Ο2 qu'à des concentrations >500μΜ. A ImM, ΓΗ2Ο2 supprimait l'activité auriculaire dans 45% des coeurs et la conduction auriculo-ventriculaire dans 66% et augmentait la formation de P-Tyr STAT3a et sa liaison à l'ADN sans modifier l'expression des gènes cibles.Conclusion: Les ROS produits par l'anoxie-réoxygénation activent STAT3a qui subit une translocation dans le noyau sans se lier à l'ADN et interagit rapidement avec des protéines de la voie RISK dans les compartiments nucléaire et cytopiasmique du coeur embryonnaire. Ce dernier, en particulier au niveau des oreillettes, se révèle très résistant au puissant stress oxydatif de l'H202 qui se différencie du stress lié à la réoxygénation en favorisant la liaison de STAT3 à l'ADN. Ces résultats originaux permettent une meilleure compréhension des mécanismes qui peuvent améliorer la récupération du coeur en développement après un épisode hypoxique intra-utérin.

The building blocks of the full body ownership illusion

Previous work has reported that it is not difficult to give people the illusion of ownership over an artificial body, providing a powerful tool for the investigation of the neural and cognitive mechanisms underlying body perception and self consciousness. We present an experimental study that uses immersive virtual reality (IVR) focused on identifying the perceptual building blocks of this illusion. We systematically manipulated visuotactile and visual sensorimotor contingencies, visual perspective, and the appearance of the virtual body in order to assess their relative role and mutual interaction. Consistent results from subjective reports and physiological measures showed that a first person perspective over a fake humanoid body is essential for eliciting a body ownership illusion. We found that the illusion of ownership can be generated when the virtual body has a realistic skin tone and spatially substitutes the real body seen from a first person perspective. In this case there is no need for an additional contribution of congruent visuotactile or sensorimotor cues. Additionally, we found that the processing of incongruent perceptual cues can be modulated by the level of the illusion: when the illusion is strong, incongruent cues are not experienced as incorrect. Participants exposed to asynchronous visuotactile stimulation can experience the ownership illusion and perceive touch as originating from an object seen to contact the virtual body. Analogously, when the level of realism of the virtual body is not high enough and/or when there is no spatial overlap between the two bodies, then the contribution of congruent multisensory and/or sensorimotor cues is required for evoking the illusion. On the basis of these results and inspired by findings from neurophysiological recordings in the monkey, we propose a model that accounts for many of the results reported in the literature.

Utilite du Remler pour eviter de surtraiter la population dite hypertendue. [Usefulness of the Remler device in avoiding overtreatment of the so-called hypertensive population]

To assess the reliability of the Remler system, a semi-automatic pressure recording device, several blood pressures were measured simultaneously by the conventional auscultatory method and by the Remler in 12 normotensive volunteers. In all situations tested both the Remler and auscultatory blood pressures were very close, thus demonstrating the reliability of this new technique. Ambulatory blood pressure recordings were then obtained with the Remler in 245 untreated patients referred for hypertension by their private physicians. Surprisingly, in close to 60% of them the average of all blood pressures recorded during usual activities was within the normal range. Since cardiovascular complications seem to correlate better with ambulatory than with office blood pressure levels, the Remler system appears particularly useful in recognizing those patients who, although hypertensive in the physician's office, remain normotensive during the day and therefore may not require antihypertensive treatment.

Transfer entropy reconstruction and labeling of neuronal connections from simulated calcium imaging

Neuronal dynamics are fundamentally constrained by the underlying structural network architecture, yet much of the details of this synaptic connectivity are still unknown even in neuronal cultures in vitro. Here we extend a previous approach based on information theory, the Generalized Transfer Entropy, to the reconstruction of connectivity of simulated neuronal networks of both excitatory and inhibitory neurons. We show that, due to the model-free nature of the developed measure, both kinds of connections can be reliably inferred if the average firing rate between synchronous burst events exceeds a small minimum frequency. Furthermore, we suggest, based on systematic simulations, that even lower spontaneous inter-burst rates could be raised to meet the requirements of our reconstruction algorithm by applying a weak spatially homogeneous stimulation to the entire network. By combining multiple recordings of the same in silico network before and after pharmacologically blocking inhibitory synaptic transmission, we show then how it becomes possible to infer with high confidence the excitatory or inhibitory nature of each individual neuron.

Sustained sleep fragmentation induces sleep homeostasis in mice.

STUDY OBJECTIVES: Sleep fragmentation (SF) is an integral feature of sleep apnea and other prevalent sleep disorders. Although the effect of repetitive arousals on cognitive performance is well documented, the effects of long-term SF on electroencephalography (EEG) and molecular markers of sleep homeostasis remain poorly investigated. To address this question, we developed a mouse model of chronic SF and characterized its effect on EEG spectral frequencies and the expression of genes previously linked to sleep homeostasis including clock genes, heat shock proteins, and plasticity-related genes. DESIGN: N/A. SETTING: Animal sleep research laboratory. PARTICIPANTS: Sixty-six C57BL6/J adult mice. INTERVENTIONS: Instrumental sleep disruption at a rate of 60/h during 14 days. MEASUREMENTS AND RESULTS: Locomotor activity and EEG were recorded during 14 days of SF followed by recovery for 2 days. Despite a dramatic number of arousals and decreased sleep bout duration, SF minimally reduced total quantity of sleep and did not significantly alter its circadian distribution. Spectral analysis during SF revealed a homeostatic drive for slow wave activity (SWA; 1-4 Hz) and other frequencies as well (4-40 Hz). Recordings during recovery revealed slow wave sleep consolidation and a transient rebound in SWA, and paradoxical sleep duration. The expression of selected genes was not induced following chronic SF. CONCLUSIONS: Chronic SF increased sleep pressure confirming that altered quality with preserved quantity triggers core sleep homeostasis mechanisms. However, it did not induce the expression of genes induced by sleep loss, suggesting that these molecular pathways are not sustainably activated in chronic diseases involving SF.

Interobserver agreement and validity of bedside 'positive signs' for functional weakness, sensory and gait disorders in conversion disorder: a pilot study.

BACKGROUND: Conversion disorder (CD) is no longer a diagnosis of exclusion. The new DSM-V criteria highlight the importance of 'positive signs' on neurological examination. Only few signs have been validated, and little is known about their reliability. OBJECTIVE: The aim was to examine the clinical value of bedside positive signs in the diagnosis of CD presenting with weakness, gait or sensory symptoms by assessing their specificity, sensitivity and their inter-rater reliability. PATIENTS AND METHODS: Standardised video recorded neurological examinations were performed in 20 consecutive patients with CD and 20 'organic' controls. Ten previously validated sensory and motor signs were grouped in a scale. Thirteen additional motor/sensory 'positive signs', 14 gait patterns and 1 general sign were assessed in a pilot validation study. In addition, two blinded independent neurologists rated the video recordings to assess the inter-rater reliability (Cohen's κ) of each sign. RESULTS: A score of ≥4/14 on the sensory motor scale showed a 100% specificity (CI 85 to 100) and a 95% sensitivity (CI 85 to 100). Among the additional tested signs, 10 were significantly more frequent in CD than controls. The interobserver agreement was acceptable for 23/38 signs (2 excellent, 10 good, 11 moderate). CONCLUSIONS: Our study confirms that six bedside 'positive signs' are highly specific for CD with good-excellent inter-rater reliability; we propose to consider them as 'highly reliable signs'. In addition 13 signs could be considered as 'reliable signs' and six further signs as 'suggestive signs' while all others should be used with caution until further validation is available.

The concept of temporal 'plus' epilepsy.

The concept of temporal 'plus' epilepsy (T+E) is not new, and a number of observations made by means of intracerebral electrodes have illustrated the complexity of neuronal circuits that involve the temporal lobe. The term T+E was used to unify and better individualize these specific forms of multilobar epilepsies, which are characterized by electroclinical features primarily suggestive of temporal lobe epilepsy, MRI findings that are either unremarkable or show signs of hippocampal sclerosis, and intracranial recordings which demonstrate that seizures arise from a complex epileptogenic network including a combination of brain regions located within the temporal lobe and over closed neighbouring structures such as the orbitofrontal cortex, the insulo-opercular region, and the temporo-parieto-occipital junction. We will review here how the term of T+E has emerged, what it means, and which practical consideration it raises.

Signaling across the synapse: a role for Wnt and Dishevelled in presynaptic assembly and neurotransmitter release

Proper dialogue between presynaptic neurons and their targets is essential for correct synaptic assembly and function. At central synapses, Wnt proteins function as retrograde signals to regulate axon remodeling and the accumulation of presynaptic proteins. Loss of Wnt7a function leads to defects in the localization of presynaptic markers and in the morphology of the presynaptic axons. We show that loss of function of Dishevelled-1 (Dvl1) mimics and enhances the Wnt7a phenotype in the cerebellum. Although active zones appear normal, electrophysiological recordings in cerebellar slices from Wnt7a/Dvl1 double mutant mice reveal a defect in neurotransmitter release at mossy fi ber–granule cell synapses. Deficiency in Dvl1 decreases, whereas exposure to Wnt increases, synaptic vesicle recycling in mossy fi bers. Dvl increases the number of Bassoon clusters, and like other components of the Wnt pathway, it localizes to synaptic sites. These fi ndings demonstrate that Wnts signal across the synapse on Dvl-expressing presynaptic terminals to regulate synaptic assembly and suggest a potential novel function for Wnts in neurotransmitter release.

Brain dynamics of meal size selection in humans.

Although neuroimaging research has evidenced specific responses to visual food stimuli based on their nutritional quality (e.g., energy density, fat content), brain processes underlying portion size selection remain largely unexplored. We identified spatio-temporal brain dynamics in response to meal images varying in portion size during a task of ideal portion selection for prospective lunch intake and expected satiety. Brain responses to meal portions judged by the participants as 'too small', 'ideal' and 'too big' were measured by means of electro-encephalographic (EEG) recordings in 21 normal-weight women. During an early stage of meal viewing (105-145ms), data showed an incremental increase of the head-surface global electric field strength (quantified via global field power; GFP) as portion judgments ranged from 'too small' to 'too big'. Estimations of neural source activity revealed that brain regions underlying this effect were located in the insula, middle frontal gyrus and middle temporal gyrus, and are similar to those reported in previous studies investigating responses to changes in food nutritional content. In contrast, during a later stage (230-270ms), GFP was maximal for the 'ideal' relative to the 'non-ideal' portion sizes. Greater neural source activity to 'ideal' vs. 'non-ideal' portion sizes was observed in the inferior parietal lobule, superior temporal gyrus and mid-posterior cingulate gyrus. Collectively, our results provide evidence that several brain regions involved in attention and adaptive behavior track 'ideal' meal portion sizes as early as 230ms during visual encounter. That is, responses do not show an increase paralleling the amount of food viewed (and, in extension, the amount of reward), but are shaped by regulatory mechanisms.

The first SEPServer event catalogue 68-MeV solar proton events observed at 1 AU in 1996-2010

SEPServer is a three-year collaborative project funded by the seventh framework programme (FP7-SPACE) of the European Union. The objective of the project is to provide access to state-of-the-art observations and analysis tools for the scientific community on solar energetic particle (SEP) events and related electromagnetic (EM) emissions. The project will eventually lead to better understanding of the particle acceleration and transport processes at the Sun and in the inner heliosphere. These processes lead to SEP events that form one of the key elements of space weather. In this paper we present the first results from the systematic analysis work performed on the following datasets: SOHO/ERNE, SOHO/EPHIN, ACE/EPAM, Wind/WAVES and GOES X-rays. A catalogue of SEP events at 1 AU, with complete coverage over solar cycle 23, based on high-energy (~68-MeV) protons from SOHO/ERNE and electron recordings of the events by SOHO/EPHIN and ACE/EPAM are presented. A total of 115 energetic particle events have been identified and analysed using velocity dispersion analysis (VDA) for protons and time-shifting analysis (TSA) for electrons and protons in order to infer the SEP release times at the Sun. EM observations during the times of the SEP event onset have been gathered and compared to the release time estimates of particles. Data from those events that occurred during the European day-time, i.e., those that also have observations from ground-based observatories included in SEPServer, are listed and a preliminary analysis of their associations is presented. We find that VDA results for protons can be a useful tool for the analysis of proton release times, but if the derived proton path length is out of a range of 1 AU < s[3 AU, the result of the analysis may be compromised, as indicated by the anti-correlation of the derived path length and release time delay from the asso ciated X-ray flare. The average path length derived from VDA is about 1.9 times the nominal length of the spiral magnetic field line. This implies that the path length of first-arriving MeV to deka-MeV protons is affected by interplanetary scattering. TSA of near-relativistic electrons results in a release time that shows significant scatter with respect to the EM emissions but with a trend of being delayed more with increasing distance between the flare and the nominal footpoint of the Earth-connected field line.

An innovative and shared methodology for event reconstruction using images in forensic science

This study presents an innovative methodology for forensic science image analysis for event reconstruction. The methodology is based on experiences from real cases. It provides real added value to technical guidelines such as standard operating procedures (SOPs) and enriches the community of practices at stake in this field. This bottom-up solution outlines the many facets of analysis and the complexity of the decision-making process. Additionally, the methodology provides a backbone for articulating more detailed and technical procedures and SOPs. It emerged from a grounded theory approach; data from individual and collective interviews with eight Swiss and nine European forensic image analysis experts were collected and interpreted in a continuous, circular and reflexive manner. Throughout the process of conducting interviews and panel discussions, similarities and discrepancies were discussed in detail to provide a comprehensive picture of practices and points of view and to ultimately formalise shared know-how. Our contribution sheds light on the complexity of the choices, actions and interactions along the path of data collection and analysis, enhancing both the researchers' and participants' reflexivity.