All-optical interrogation of neural circuits in behaving mice.

Recent advances combining two-photon calcium imaging and two-photon optogenetics with computer-generated holography now allow us to read and write the activity of large populations of neurons in vivo at cellular resolution and with high temporal resolution. Such 'all-optical' techniques enable experimenters to probe the effects of functionally defined neurons on neural circuit function and behavioral output with new levels of precision. This greatly increases flexibility, resolution, targeting specificity and throughput compared with alternative approaches based on electrophysiology and/or one-photon optogenetics and can interrogate larger and more densely labeled populations of neurons than current voltage imaging-based implementations. This protocol describes the experimental workflow for all-optical interrogation experiments in awake, behaving head-fixed mice. We describe modular procedures for the setup and calibration of an all-optical system (~3 h), the preparation of an indicator and opsin-expressing and task-performing animal (~3-6 weeks), the characterization of functional and photostimulation responses (~2 h per field of view) and the design and implementation of an all-optical experiment (achievable within the timescale of a normal behavioral experiment; ~3-5 h per field of view). We discuss optimizations for efficiently selecting and targeting neuronal ensembles for photostimulation sequences, as well as generating photostimulation response maps from the imaging data that can be used to examine the impact of photostimulation on the local circuit. We demonstrate the utility of this strategy in three brain areas by using different experimental setups. This approach can in principle be adapted to any brain area to probe functional connectivity in neural circuits and investigate the relationship between neural circuit activity and behavior.

Is current body temperature measurement practice fit-for-purpose?

There has been a marked rise in the number of avoidable deaths in health services around the world. At the same time there has been a growing increase in antibiotic resistant so-called "superbugs." We examine here the potential role of body temperature measurement in these adverse trends. Electronic based thermometers have replaced traditional mercury (and other liquid-in-glass type) thermometers for reasons of safety rather than superiority. Electronic thermometers are in general less robust from a measurement perspective than their predecessors. We illustrate the implications of unreliable temperature measurement on the diagnosis and management of disease, including COVID-19, through statistical calculations. Since a return to mercury thermometers is both undesirable and impractical, we call for better governance in the current practice of clinical thermometry to ensure the traceability and long-term accuracy of electronic thermometers and discuss how this could be achieved.

Targeted Activation of Hippocampal Place Cells Drives Memory-Guided Spatial Behavior.

The hippocampus is crucial for spatial navigation and episodic memory formation. Hippocampal place cells exhibit spatially selective activity within an environment and have been proposed to form the neural basis of a cognitive map of space that supports these mnemonic functions. However, the direct influence of place cell activity on spatial navigation behavior has not yet been demonstrated. Using an 'all-optical' combination of simultaneous two-photon calcium imaging and two-photon optogenetics, we identified and selectively activated place cells that encoded behaviorally relevant locations in a virtual reality environment. Targeted stimulation of a small number of place cells was sufficient to bias the behavior of animals during a spatial memory task, providing causal evidence that hippocampal place cells actively support spatial navigation and memory.

Do Helpful Mothers Help? Effects of Maternal Scaffolding and Infant Engagement on Cognitive Performance.

Infants are highly social and much early learning takes place in a social context during interactions with caregivers. Previous research shows that social scaffolding - responsive parenting and joint attention - can confer benefits for infants' long-term development and learning. However, little previous research has examined whether dynamic (moment-to-moment) adaptations in adults' social scaffolding are able to produce immediate effects on infants' performance. Here we ask whether infants' success on an object search task is more strongly influenced by maternal behavior, including dynamic changes in response behavior, or by fluctuations in infants' own engagement levels. Thirty-five mother-infant dyads (infants aged 10.8 months, on average) participated in an object search task that was delivered in a naturalistic manner by the child's mother. Measures of maternal responsiveness (teaching duration; sensitivity) and infant engagement (engagement score; visual attention) were assessed. Mothers varied their task delivery trial by trial, but neither measure of maternal responsiveness significantly predicted infants' success in performing the search task. Rather, infants' own level of engagement was the sole significant predictor of accuracy. These results indicate that while parental scaffolding is offered spontaneously (and is undoubtedly crucial for development), in this context children's endogenous engagement proved to be a more powerful determinant of task success. Future work should explore this interplay between parental and child-internal factors in other learning and social contexts.

Parental neural responsivity to infants' visual attention: How mature brains influence immature brains during social interaction.

Almost all attention and learning-in particular, most early learning-take place in social settings. But little is known of how our brains support dynamic social interactions. We recorded dual electroencephalography (EEG) from 12-month-old infants and parents during solo play and joint play. During solo play, fluctuations in infants' theta power significantly forward-predicted their subsequent attentional behaviours. However, this forward-predictiveness was lower during joint play than solo play, suggesting that infants' endogenous neural control over attention is greater during solo play. Overall, however, infants were more attentive to the objects during joint play. To understand why, we examined how adult brain activity related to infant attention. We found that parents' theta power closely tracked and responded to changes in their infants' attention. Further, instances in which parents showed greater neural responsivity were associated with longer sustained attention by infants. Our results offer new insights into how one partner influences another during social interaction.

Infants' visual sustained attention is higher during joint play than solo play: is this due to increased endogenous attention control or exogenous stimulus capture?

Previous research has suggested that when a social partner, such as a parent, pays attention to an object, this increases the attention that infants pay to that object during spontaneous, naturalistic play. There are two contrasting reasons why this might be: first, social context may influence increases in infants' endogenous (voluntary) attention control; second, social settings may offer increased opportunities for exogenous attentional capture. To differentiate these possibilities, we compared 12-month-old infants' naturalistic attention patterns in two settings: Solo Play and Joint Play with a social partner (the parent). Consistent with previous research, we found that infants' look durations toward play objects were longer during Joint Play, and that moments of inattentiveness were fewer, and shorter. Follow-up analyses, conducted to differentiate the two above-proposed hypotheses, were more consistent with the latter hypothesis. We found that infants' rate of change of attentiveness was faster during Joint Play than Solo Play, suggesting that internal attention factors, such as attentional inertia, may influence looking behaviour less during Joint Play. We also found that adults' attention forwards-predicted infants' subsequent attention more than vice versa, suggesting that adults' behaviour may drive infants' behaviour. Finally, we found that mutual gaze did not directly facilitate infant attentiveness. Overall, our results suggest that infants spend more time attending to objects during Joint Play than Solo Play, but that these differences are more likely attributable to increased exogenous attentional scaffolding from the parent during social play, rather than to increased endogenous attention control from the infant.

A multi-institutional dosimetry audit of rotational intensity-modulated radiotherapy.

BACKGROUND: Rotational IMRT (VMAT and Tomotherapy) has now been implemented in many radiotherapy centres. An audit to verify treatment planning system modelling and treatment delivery has been undertaken to ensure accurate clinical implementation. MATERIAL AND METHODS: 34 institutions with 43 treatment delivery systems took part in the audit. A virtual phantom planning exercise (3DTPS test) and a clinical trial planning exercise were planned and independently measured in each institution using a phantom and array combination. Point dose differences and global gamma index (γ) were calculated in regions corresponding to PTVs and OARs. RESULTS: Point dose differences gave a mean (±sd) of 0.1±2.6% and 0.2±2.0% for the 3DTPS test and clinical trial plans, respectively. 34/43 planning and delivery combinations achieved all measured planes with >95% pixels passing γ<1 at 3%/3mm and rose to 42/43 for clinical trial plans. A statistically significant difference in γ pass rates (p<0.01) was seen between planning systems where rotational IMRT modelling had been designed for the manufacturer's own treatment delivery system and those designed independently of rotational IMRT delivery. CONCLUSIONS: A dosimetry audit of rotational radiotherapy has shown that TPS modelling and delivery for rotational IMRT can achieve high accuracy of plan delivery.

Comparison of air kerma standards of LNE-LNHB and NPL for 192Ir HDR brachytherapy sources: EUROMET project no 814.

An indirect comparison has been made in the air kerma standards for high dose rate (HDR) 192Ir brachytherapy sources at the Laboratoire National Henri Becquerel (LNHB) and the National Physical Laboratory (NPL). The measurements were carried out at both laboratories between November and December 2004. The comparison was based on measurements using well-type transfer ionization chambers and two different source types, Nucletron microSelectron HDR Classic and version 2. The results show the reported calibration coefficients to agree within 0.47% to 0.63%, which is within the overall standard uncertainty of 0.65% reported by both laboratories at the time of this comparison. Following this comparison, some of the NPL primary standard correction factors were re-evaluated resulting in a change of +0.17% in the overall correction factor. The new factor was implemented in May 2006. Applying the revised chamber factor to the measurements reported in this comparison report will reduce the difference between the two standards by 0.17%.