Functional 2‐photon microscopy is a key technology for imaging neuronal activity. The recorded image sequences, however, can contain non‐rigid movement artifacts which requires high‐accuracy movement ...correction. Variational optical flow (OF) estimation is a group of methods for motion analysis with established performance in many computer vision areas. However, it has yet to be adapted to the statistics of 2‐photon neuroimaging data. In this work, we present the motion compensation method Flow‐Registration that outperforms previous alignment tools and allows to align and reconstruct even low signal‐to‐noise ratio 2‐photon imaging data and is able to compensate high‐divergence displacements during local drug injections. The method is based on statistics of such data and integrates previous advances in variational OF estimation. Our method is available as an easy‐to‐use ImageJ/FIJI plugin as well as a MATLAB toolbox with modular, object oriented file IO, native multi‐channel support and compatibility with existing 2‐photon imaging suites.
We present the non‐parametric, high‐accuracy and fast motion compensation method Flow‐Registration that can compensate challenging 2‐Photon neuroimaging videos which are contaminated with large and or high divergence displacements. Our method is publicly available as a MATLAB toolbox and ImageJ/FIJI plugin on GitHub: https://github.com/phflot/flow_registration
Summary This study investigated variations in salivary levels of cortisol and dehydroepiandrosterone (DHEA) in a prolonged stressful situation (a two-week teaching practice). Thirty-three women for ...whom a two-week teaching practice at a kindergarten was scheduled were asked to collect saliva samples at awakening, 30 min after awakening, and bedtime at four time points: two weeks before the practice, the first week of the practice, the second week of the practice, and a few days after the practice. In addition, they completed questionnaires for assessing perceived stress and subjective moods on each day. A linear mixed model indicated that cortisol levels significantly increased during the first and second week of the practice compared with those before and after the practice period, and that DHEA levels significantly decreased after the practice period compared with those at the other time points. Further, cortisol awakening response after the practice period significantly reduced compared with that at the other time points. Scores of perceived stress and negative moods were also higher during the practice period. This study showed that prolonged stress affected cortisol and DHEA secretion during as well as after the stress period.
Little is known about the real-time cellular dynamics triggered by endogenous catecholamine release despite their importance in brain functions. To address this issue, we expressed channelrhodopsin ...in locus coeruleus neurons and protein kinase-A activity biosensors in cortical pyramidal neurons and combined two-photon imaging of biosensors with photostimulation of locus coeruleus cortical axons, in acute slices and in vivo. Burst photostimulation of axons for 5–10 s elicited robust, minutes-lasting kinase-A activation in individual neurons, indicating that a single burst firing episode of synchronized locus coeruleus neurons has rapid and lasting effects on cortical network. Responses were mediated by β1 adrenoceptors, dampened by co-activation of α2 adrenoceptors, and dramatically increased upon inhibition of noradrenaline reuptake transporter. Dopamine receptors were not involved, showing that kinase-A activation was due to noradrenaline release. Our study shows that noradrenergic transmission can be characterized with high spatiotemporal resolution in brain slices and in vivo using optogenetic tools.
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•We report all-optogenetic stimulation and imaging of NA transmission in cortex•NA fiber stimulation elicits robust PKA activation in neurons, in slice and in vivo•PKA activation involves both β1 and α2 adrenoceptors but not dopamine receptors•Noradrenaline reuptake transporter activity greatly limits response intensity
Optical Imaging; Neuroscience; Cell Biology
Aim
To verify the reliability and validity of Multidimensional Measure of Leader–Member Exchange Japanese version (LMX‐MDM‐J) for staff nurses.
Background
Staff nurses who have a high‐quality ...relationship with nurse managers tend to have low turnover intention. However, the validity and reliability of LMX‐MDM Japanese version have not been confirmed.
Methods
Staff nurses (n = 870) working in wards of four Japanese hospitals were surveyed using self‐administered questionnaires. Reliability was tested by internal consistency. Validity was tested by the confirmatory factor analysis for construct validity and relationships with external criteria for criterion‐related validity.
Results
There were 450 valid responses. Cronbach's α coefficients of the overall scale and each dimension were 0.97 and 0.86–0.95, respectively. The goodness‐of‐fit indices of the confirmatory factor analysis showed CFI = 0.981 and RMSEA = 0.076. Correlation coefficients with external criteria were 0.57 for job satisfaction, 0.80 for relationship satisfaction with the nurse manager and −0.36 for turnover intention (all p < .001).
Conclusion
The reliability and validity of LMX‐MDM‐J were determined to be adequate for staff nurses.
Implications for Nursing Management
LMX‐MDM‐J can accurately measure the quality of the dyadic relationship between nurse managers and staff nurses. This measurement indicates whether nurse managers are providing leadership.
Recent reports point to critical roles of glutamate receptor subunit delta2 (GluD2) at excitatory synapses and link GluD1 gene alteration to schizophrenia but the expression patterns of these ...subunits in the brain remain almost uncharacterized. We examined the distribution of GluD1–2 mRNAs and proteins in the adult rodent brain, focusing mainly on GluD1. In situ hybridization revealed widespread neuronal expression of the GluD1 mRNA, with higher levels occurring in several forebrain regions and lower levels in cerebellum. Quantitative RT-PCR assessed differential GluD1 expression in cortex and cerebellum, and revealed GluD2 expression in cortex, albeit at markedly lower level than in cerebellum. Likewise, a high GluD1/GluD2 mRNA ratio was observed in cortex and a low ratio in cerebellum. GluD1 and GluD2 mRNAs were co-expressed in single cortical and hippocampal neurons, with a large predominance of GluD1. Western blots using GluD1- and GluD2-specific antibodies showed expression of both subunits in various brain structures, but not in non-nervous tissues examined. Both delta subunits were upregulated during postnatal development. Widespread neuronal expression of the GluD1 protein was confirmed using immunohistochemistry. Examination at the electron microscopic level in the hippocampus revealed that GluD1 was mainly localized at postsynaptic density of excitatory synapses on pyramidal cells. Control experiments performed using mice carrying deletion of the GluD1- or the GluD2-encoding gene confirmed the specificity of the present mRNA and protein analyses. Our results support a role for the delta family of glutamate receptors at excitatory synapses in neuronal networks throughout the adult brain.
•Salivary IL-6 levels are elevated for 20min following acute psychosocial stress.•Individuals with sustained elevation of IL-6 levels showed a lower cortisol response.•A greater IL-6 response was ...associated with a higher heart rate during the task.
Although interleukin-6 (IL-6) has been investigated frequently in stress research, knowledge regarding the biological processes of IL-6 in association with psychosocial stress remains incomplete. This study focused on salivary IL-6 and reports its temporal variation and biological correlates following acute psychosocial stress. Fifty healthy young adults (39 male and 11 female students) were subjected to the psychosocial stress test ‘Trier Social Stress Test’ (TSST), wherein the participants were asked to deliver a speech and perform a mental arithmetic task in front of 2 audiences. Collection of saliva samples, measurement of heart rate, and assessment of negative moods by visual analogue scales were conducted before, during, and after TSST. Salivary IL-6 levels increased by approximately 50% in response to the TSST and remained elevated for 20min after the stress tasks were completed. Cluster analyses revealed that individuals with sustained elevation of IL-6 levels following the TSST exhibited a lower cortisol response compared to individuals with lower IL-6 levels. In the correlation analyses, a greater IL-6 response was associated with a higher heart rate during the mental arithmetic task (r=.351, p<.05) and with a lower cortisol response (r=−.302, p<.05). This study demonstrates that salivary IL-6 levels are elevated for a relatively long period following acute psychosocial stress, and suggests that sympathetic activity and cortisol secretion are involved in elevation of salivary IL-6 levels.
Pain and itch are closely related sensations, yet qualitatively quite distinct. Despite recent advances in brain imaging techniques, identifying the differences between pain and itch signals in the ...brain cortex is difficult due to continuous temporal and spatial changes in the signals. The high spatial resolution of positron emission tomography (PET) and functional magnetic resonance imaging (fMRI) has substantially advanced research of pain and itch, but these are uncomfortable because of expensiveness, importability and the limited operation in the shielded room. Here, we used near infrared spectroscopy (NIRS), which has more conventional usability. NIRS can be used to visualize dynamic changes in oxygenated hemoglobin and deoxyhemoglobin concentrations in the capillary networks near activated neural circuits in real-time as well as fMRI. We observed distinct activation patterns in the frontal cortex for acute pain and histamine-induced itch. The prefrontal cortex exhibited a pain-related and itch-related activation pattern of blood flow in each subject. Although it looked as though that activation pattern for pain and itching was different in each subject, further cross correlation analysis of NIRS signals between each channels showed an overall agreement with regard to prefrontal area involvement. As a result, pain-related and itch-related blood flow responses (delayed responses in prefrontal area) were found to be clearly different between pain (τ = +18.7 sec) and itch (τ = +0.63 sec) stimulation. This is the first pilot study to demonstrate the temporal and spatial separation of a pain-induced blood flow and an itch-induced blood flow in human cortex during information processing.
Recent studies have demonstrated that depletion of amino acids results in the induction of several genes and that a genomic cis-element termed amino acid response element (AARE) is required for the ...induction. System xc- is an anionic amino acid transport system highly specific for cystine and glutamate, and its activity is known to be induced by cystine deprivation. This transporter is composed of two protein components, xCT and 4F2 heavy chain, and xCT is thought to mediate the transport activity. In the present study, the molecular mechanism for the induction of xCT by amino acid deprivation has been investigated. In mouse NIH3T3 cells, the activity of system xc- and xCT mRNA is induced not only by deprivation of cystine but also by deprivation of other amino acids. Two AAREs, each located in the opposite direction with an intervening sequence, were found in the 5′-flanking region of the mouse xCT gene. Promoter analysis revealed that both AAREs were necessary for the maximal induction of xCT mRNA in response to the amino acid deprivation. Glucose deprivation had no effect on the induction of the activity of system xc-. Electrophoretic mobility shift assay showed that ATF4, but not ATF2, is involved in the amino acid control of xCT expression. These results demonstrate that xCT is a new member of the proteins whose transcriptional control by the amino acid deprivation is mediated by AARE.
A cross-sectional survey was conducted to simultaneously evaluate sleep quality, duration, and phase in school-aged children and correlations between each dimension of sleep and daytime sleepiness ...were comprehensively examined. A cross-sectional survey was conducted with school-aged children enrolled in four public elementary schools in Joetsu city, Niigata prefecture in Japan (
n
= 1683). Among the collected responses (
n
= 1290), 1134 valid responses (547 boys and 587 girls) were analyzed (valid response rate was 87.90%). Data on daytime sleepiness, sleep quality (problems in sleeping at night), sleep duration (the average sleeping time during a week), and sleep phase (sleep timing: bedtime and rising time on weekdays, and sleep regularity: differences in bedtime and rising time between on weekdays and weekends) were collected. The results of multivariate logistic regression analysis indicated that the following dimensions were significantly correlated with daytime sleepiness: the decline in sleep quality adjusted odds ratio (AOR) = 2.62, 95% confidence interval (CI) = 1.71–4.00, bedtime after 21:30 on weekdays (AOR = 1.58, 95% CI = 1.15–2.18), bedtime delay on weekends, compared to weekdays (AOR = 1.75, 95% CI = 1.27–2.41), and bedtime advance on weekends, compared to weekdays (AOR = 3.33, 95% CI = 1.78–6.20). Sleep dimensions that significantly affected daytime sleepiness in school-aged children are sleep quality, bedtime-timing, and regularity of bedtime. It is important to detect problems in night sleep and establish treatments, as well as to provide support for early bedding on weekdays and for a regular bedtime both on weekdays and on weekends to prevent daytime sleepiness in school-aged children.
Objectives To develop a stressor scale for Japanese overseas residents that takes their general and specific circumstances into consideration. Methods In Study 1, an original version of the Stressor ...Scale for Japanese Overseas Residents - United Kingdom version (SSJR-UK) consisting of 47 items was drafted based on the Daily Hassles Scale and 13 items extracted using the KJ Method. Next, we administered it to Japanese residents in the UK (n = 369) to assess its reliability and construct validity. In Study 2, a subset of the UK group (n = 100) was compared to three other groups: Nordic countries (n = 65), Southeast Asia (n = 61), and Japan (n = 148) to identify stressors specific to life in the UK. Results Factor analysis was conducted using promax rotation and the following six factors were identified: daily life circumstances, psychological pressure, sense of fulfilment, health and future of the family, interpersonal relationships, and oral communication. Overall Cronbach’s alpha was .90, which provides evidence of the scale’s high internal consistency. Pearson product-moment correlation coefficients between each SSJR-UK factor scores and the CES-D were .28-.58 (p < .01) and between SSJR-UK factor scores and the SRS18 were .38-.67 (p <.01), which are indicative of the construct validity of the scale. The discriminability of the scale indicated that only the overall score and scores for the factors specific to conditions in the UK were significantly higher than in the other countries: F (4,370) = 9.19 (p < .001) and F (4,370) = 16.48 (p < .001), respectively. Conclusion The SSJR-UK has sufficient reliability and validity to measure the stressors of Japanese residents in the UK.
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