Hdac1-deficiency affects the cell cycle axis Cdc25-Cdk1 causing impaired G2/M phase progression and reduced cardiomyocyte proliferation in zebrafish.

Zebrafish have the ability to fully regenerate their hearts after injury since cardiomyocytes subsequently dedifferentiate, re-enter cell cycle, and proliferate to replace damaged myocardial tissue. Recent research identified the reactivation of dormant developmental pathways during cardiac regeneration in adult zebrafish, suggesting pro-proliferative pathways important for developmental heart growth to be also critical for regenerative heart growth after injury. Histone deacetylase 1 (Hdac1) was recently shown to control both, embryonic as well as adult regenerative cardiomyocyte proliferation in the zebrafish model. Nevertheless, regulatory pathways controlled by Hdac1 are not defined yet. By analyzing RNA-seq-derived transcriptional profiles of the Hdac1-deficient zebrafish mutant baldrian, we here identified DNA damage response (DDR) pathways activated in baldrian mutant embryos. Surprisingly, although the DDR signaling pathway was transcriptionally activated, we found the complete loss of protein expression of the known DDR effector and cell cycle inhibitor p21. Consequently, we observed an upregulation of the p21-downstream target Cdk2, implying elevated G1/S phase transition in Hdac1-deficient zebrafish hearts. Remarkably, Cdk1, another p21-but also Cdc25-downstream target was downregulated. Here, we found the significant downregulation of Cdc25 protein expression, explaining reduced Cdk1 levels and suggesting impaired G2/M phase progression in Hdac1-deficient zebrafish embryos. To finally prove defective cell cycle progression due to Hdac1 loss, we conducted Cytometer-based cell cycle analyses in HDAC1-deficient murine HL-1 cardiomyocytes and indeed found impaired G2/M phase transition resulting in defective cardiomyocyte proliferation. In conclusion, our results suggest a critical role of Hdac1 in maintaining both, regular G1/S and G2/M phase transition in cardiomyocytes by controlling the expression of essential cell cycle regulators such as p21 and Cdc25.

[Knowledge facilitates health: the "Fit in Health" program for strengthening the health literacy of students]. / Wissen schafft Gesundheit: Das Programm "Fit in Gesundheitsfragen" zur Stärkung der Gesundheitskompetenz von Schülerinnen und Schülern.

In Germany, more than half of the population has low health literacy. These people have difficulties in finding and classifying health-related information and adapting it to their own situation. Among them are many young people, highlighting the importance of interventions early in life to promote health literacy.This is where the "Fit in Health" program of the Health Information Services of the Helmholtz Munich Research Center and the German Cancer Research Center starts. The objective is to contribute to the promotion of different dimensions of health literacy among children and adolescents. To this end, innovative training formats for teachers and teaching materials for students at secondary levels I and II are being designed and evaluated. The major diseases cancer and diabetes mellitus are used as models to provide basic knowledge on their pathogenesis, prevention, treatment, and research. In addition, information about the structures of the healthcare system and materials to strengthen health literacy are provided. An accompanying evaluation collects indicator data on coverage and acceptance of the measures.Since 2018, the program has published 46 teaching materials and three course readers with background knowledge for teachers. Furthermore, 50 training courses have been held, with approximately 1600 teachers and multipliers taking part in face-to-face and online events by the end of 2021. More than 90% of the participants gave the respective events very good or good evaluations. Around 80% of the participants said they would like to include the topics taught in their lessons. Further expansion of the offers is planned. Testing of a set of materials with respect to the gain of health literacy in a sample of students is currently being prepared.

Correction: Stimulation of the left dorsolateral prefrontal cortex with slow rTMS enhances verbal memory formation.

[This corrects the article DOI: 10.1371/journal.pbio.3001363.].

Stimulation of the left dorsolateral prefrontal cortex with slow rTMS enhances verbal memory formation.

Encoding of episodic memories relies on stimulus-specific information processing and involves the left prefrontal cortex. We here present an incidental finding from a simultaneous EEG-TMS experiment as well as a replication of this unexpected effect. Our results reveal that stimulating the left dorsolateral prefrontal cortex (DLPFC) with slow repetitive transcranial magnetic stimulation (rTMS) leads to enhanced word memory performance. A total of 40 healthy human participants engaged in a list learning paradigm. Half of the participants (N = 20) received 1 Hz rTMS to the left DLPFC, while the other half (N = 20) received 1 Hz rTMS to the vertex and served as a control group. Participants receiving left DLPFC stimulation demonstrated enhanced memory performance compared to the control group. This effect was replicated in a within-subjects experiment where 24 participants received 1 Hz rTMS to the left DLPFC and vertex. In this second experiment, DLPFC stimulation also induced better memory performance compared to vertex stimulation. In addition to these behavioural effects, we found that 1 Hz rTMS to DLPFC induced stronger beta power modulation in posterior areas, a state that is known to be beneficial for memory encoding. Further analysis indicated that beta modulations did not have an oscillatory origin. Instead, the observed beta modulations were a result of a spectral tilt, suggesting inhibition of these parietal regions. These results show that applying 1 Hz rTMS to DLPFC, an area involved in episodic memory formation, improves memory performance via modulating neural activity in parietal regions.

Transcranial ultrasound stimulation in humans is associated with an auditory confound that can be effectively masked.

BACKGROUND: Transcranial ultrasound stimulation (TUS) is emerging as a potentially powerful, non-invasive technique for focal brain stimulation. Recent animal work suggests, however, that TUS effects may be confounded by indirect stimulation of early auditory pathways. OBJECTIVE: We aimed to investigate in human participants whether TUS elicits audible sounds and if these can be masked by an audio signal. METHODS: In 18 healthy participants, T1-weighted magnetic resonance brain imaging was acquired for 3D ultrasound simulations to determine optimal transducer placements and source amplitudes. Thermal simulations ensured that temperature rises were <0.5 °C at the target and <3 °C in the skull. To test for non-specific auditory activation, TUS (500 kHz, 300 ms burst, modulated at 1 kHz with 50% duty cycle) was applied to primary visual cortex and participants were asked to distinguish stimulation from non-stimulation trials. EEG was recorded throughout the task. Furthermore, ex-vivo skull experiments tested for the presence of skull vibrations during TUS. RESULTS: We found that participants can hear sound during TUS and can distinguish between stimulation and non-stimulation trials. This was corroborated by EEG recordings indicating auditory activation associated with TUS. Delivering an audio waveform to participants through earphones while TUS was applied reduced detection rates to chance level and abolished the TUS-induced auditory EEG signal. Ex vivo skull experiments demonstrated that sound is conducted through the skull at the pulse repetition frequency of the ultrasound. CONCLUSION: Future studies using TUS in humans need to take this auditory confound into account and mask stimulation appropriately.

Probing the causal involvement of dlPFC in directed forgetting using rTMS-A replication study.

The forgetting of previously remembered information has, for a long time, been explained by purely passive processes. This viewpoint has been challenged by the finding that humans show worse memory for specific items that they have been instructed to forget. The dorsolateral prefrontal cortex has, through imaging, lesion and brain stimulation studies, been implied in controlling such active forgetting processes. In this study, we attempted to solidify evidence for such a causal role of the dlPFC in directed forgetting by replicating an existing rTMS study (Hanslmayr S, 2012) in a preregistered within-participant design. We stimulated participants at the dlPFC (BA9) or vertex using 45s of 1Hz rTMS after instructions to forget previously remembered words in a list-method directed forgetting paradigm and tested for effects on the amount of forgotten information. Contrary to the study we were attempting to replicate, no significant increase in forgetting under dlPFC stimulation was found in our participants. However, when combining our results with the study we were attempting to replicate, dlPFC stimulation led to significantly increased directed forgetting in both studies combined. We further explored if the rTMS parameters used here and in earlier work (Hanslmayr S, 2012) influenced inhibitory processing at their time of delivery or in a more persistent manner. Unaltered incongruency and negative priming effects in a Stroop task conducted directly after stimulation suggests that our rTMS stimulation did not continue to influence inhibitory processing after the time of stimulation. As the combined evidence for increased directed forgetting due to rTMS dlPFC stimulation is still quite weak, additional replications are necessary to show that directed forgetting is indeed causally driven by an active prefrontal process.

On the effectiveness of event-related beta tACS on episodic memory formation and motor cortex excitability.

BACKGROUND: Transcranial alternating current stimulation (tACS) is widely used to entrain or modulate brain oscillations in order to investigate causal relationships between oscillations and cognition. OBJECTIVE: In a series of experiments we here addressed the question of whether event-related, transient tACS in the beta frequency range can be used to entrain beta oscillations in two different domains: episodic memory formation and motor cortex excitability. METHODS: In experiments 1 and 2, 72 healthy human participants engaged in an incidental encoding task of verbal and non-verbal material while receiving tACS to the left and right inferior frontal gyrus (IFG) at 6.8 Hz, 10.7 Hz, 18.5 Hz, 30 Hz, 48 Hz and sham stimulation for 2s during stimulus presentation. In experiment 3, tACS was administered for 10s to M1 at the individual motor beta frequency of eight subjects. We investigated the relationship between the size of TMS induced MEPs and tACS phase. RESULTS: Beta tACS did not affect memory performance compared to sham stimulation in experiments 1 and 2. Likewise, in experiment 3, MEP size was not modulated by the tACS phase. CONCLUSIONS: Our findings suggest that event-related, transient tACS in the beta frequency range cannot be used to modulate the formation of episodic memories or motor cortex excitability. These null-results question the effectiveness of event-related tACS to entrain beta oscillations and modulate cognition.

Probing the causal role of prestimulus interregional synchrony for perceptual integration via tACS.

The phase of prestimulus oscillations at 7-10 Hz has been shown to modulate perception of briefly presented visual stimuli. Specifically, a recent combined EEG-fMRI study suggested that a prestimulus oscillation at around 7 Hz represents open and closed windows for perceptual integration by modulating connectivity between lower order occipital and higher order parietal brain regions. We here utilized brief event-related transcranial alternating current stimulation (tACS) to specifically modulate this prestimulus 7 Hz oscillation, and the synchrony between parietal and occipital brain regions. To this end we tested for a causal role of this particular prestimulus oscillation for perceptual integration. The EEG was acquired at the same time allowing us to investigate frequency specific after effects phase-locked to stimulation offset. On a behavioural level our results suggest that tACS did modulate perceptual integration, however, in an unexpected manner. On an electrophysiological level our results suggest that brief tACS does induce oscillatory entrainment, as visible in frequency specific activity phase-locked to stimulation offset. Together, our results do not strongly support a causal role of prestimulus 7 Hz oscillations for perceptual integration. However, our results suggest that brief tACS is capable of modulating oscillatory activity in a temporally sensitive manner.

Episodic Memory Retrieval Functionally Relies on Very Rapid Reactivation of Sensory Information.

Episodic memory retrieval is assumed to rely on the rapid reactivation of sensory information that was present during encoding, a process termed "ecphory." We investigated the functional relevance of this scarcely understood process in two experiments in human participants. We presented stimuli to the left or right of fixation at encoding, followed by an episodic memory test with centrally presented retrieval cues. This allowed us to track the reactivation of lateralized sensory memory traces during retrieval. Successful episodic retrieval led to a very early (∼100-200 ms) reactivation of lateralized alpha/beta (10-25 Hz) electroencephalographic (EEG) power decreases in the visual cortex contralateral to the visual field at encoding. Applying rhythmic transcranial magnetic stimulation to interfere with early retrieval processing in the visual cortex led to decreased episodic memory performance specifically for items encoded in the visual field contralateral to the site of stimulation. These results demonstrate, for the first time, that episodic memory functionally relies on very rapid reactivation of sensory information. SIGNIFICANCE STATEMENT: Remembering personal experiences requires a "mental time travel" to revisit sensory information perceived in the past. This process is typically described as a controlled, relatively slow process. However, by using electroencephalography to measure neural activity with a high time resolution, we show that such episodic retrieval entails a very rapid reactivation of sensory brain areas. Using transcranial magnetic stimulation to alter brain function during retrieval revealed that this early sensory reactivation is causally relevant for conscious remembering. These results give first neural evidence for a functional, preconscious component of episodic remembering. This provides new insight into the nature of human memory and may help in the understanding of psychiatric conditions that involve the automatic intrusion of unwanted memories.

Differences in LEA-like 11-24 gene expression in desiccation tolerant and sensitive species of Linderniaceae are due to variations in gene promoter sequences.

Many desiccation induced late embryogenesis abundant (LEA) protein encoding genes have been identified from Craterostigma plantagineum Hochst. In the desiccation tolerant plants C. plantagineum (Cp) and Lindernia brevidens Skan (Lb) transcripts encoding LEA-like 11-24 protein are abundantly expressed during desiccation whereas in Lindernia subracemosa De Wild. (Ls), a desiccation sensitive plant, the LEA-like 11-24 transcripts are expressed at a low level. Since promoters determine gene expression, a comparative promoter analysis was carried out to decipher the underlying mechanisms of differential gene expression. Two transient transformation methods (particle bombardment and optimised Agrobacterium co-cultivation) were used to analyse the promoter activities of the Cp, Lb and Ls LEA-like 11-24 gene in homologous and heterologous systems. Minimal promoters were isolated from all three species and their promoter activities were assessed in response to mannitol or ABA. Particle bombardment or Agrobacterium co-cultivation yielded similar results. Site-directed mutagenesis was used to identify which cis-acting elements in the LEA-like 11-24 promoter fragments are crucial during mannitol and ABA treatments. The presence of these promoter cis-elements explains the differences in transcript abundance in the desiccation tolerant and desiccation sensitive species. Results indicated the importance of the drought responsive elements (DRE) element for promoter activity.