Protocol for microfluidic single-cell cultivation and live-cell imaging of Chinese hamster ovary suspension cell lines.

Microfluidic single-cell cultivation (MSCC) is a powerful tool for investigating the cellular behavior of various cell types at the single-cell level. Here, we present a protocol specifically developed for the reliable and reproducible MSCC of industrially relevant Chinese hamster ovary (CHO) suspension cell lines. We summarize critical experimental steps from the initial seed train up to the final MSCC experiment, with a special focus on pre-culture management and medium preparation, device inoculation, and the establishment of a constant medium perfusion.

Social participation in the city: exploring the moderating effect of walkability on the associations between active mobility, neighborhood perceptions, and social activities in urban adults.

BACKGROUND: Living in urban environments is associated with several health risks (e.g., noise, and air pollution). However, there are also beneficial aspects such as various opportunities for social activities, which might increase levels of social participation and (physically) active mobility that in turn have positive effects on health and well-being. However, how aspects of the environment, active mobility, and social participation are associated is not well established. This study investigates the moderating effect of low vs. high walkability neighborhoods on the associations between active mobility, and social participation and integrates individuals' subjective perception of the neighborhood environment they are living in. METHODS: Cross-sectional data from 219 adults (48% female, mean age = 46 ± 3.8 years) from 12 urban neighborhoods (six low, six high walkability) were analyzed: First, social participation, active mobility, and subjective neighborhood perceptions were compared between people living in a low vs. high walkability neighborhood via t-tests. Second, multigroup path analyses were computed to explore potential differences in the associations between these variables in low vs. high walkability neighborhoods. RESULTS: Social participation, active mobility, and subjective neighborhood perceptions didn't differ in low vs. high walkability neighborhoods (p: 0.37 - 0.71). Active mobility and subjective neighborhood perceptions were significantly stronger related to social participation in low vs. high walkability neighborhoods (active mobility in low: ß = 0.35, p < .01 vs. high: ß = 0.09, p = .36; subjective neighborhood perceptions in low: ß = 0.27, p < .01 vs. high: ß = 0.15, p = .18). CONCLUSIONS: Despite living in neighborhoods with objectively different walkability, participants rated social participation and active mobility equally and perceived their neighborhoods similarly. However, zooming into the interrelations of these variables reveals that social participation of residents from low walkability neighborhoods depends stronger on active mobility and perceiving the environment positively. Positive perceptions of the environment and active mobility might buffer the objectively worse walkability. Future research should focus on underlying mechanisms and determinants of subjective neighborhood perceptions and active mobility, especially in low walkability neighborhoods.

Scoping review: longitudinal effects of the COVID-19 pandemic on child and adolescent mental health.

The COVID-19 pandemic and associated containment measures have massively changed the daily lives of billions of children and adolescents worldwide. To investigate the global longitudinal effects on various mental health outcomes over a period of 1.5 years, we conducted a scoping review in accordance with the guidelines of the Preferred Reporting Items for Systematic Reviews and Meta-Analyses extension for Scoping Reviews (PRISMA-ScR). We included the peer-reviewed articles from PubMed, Web of Science, and APA PsycInfo that were published between December 2019 and December 2021, followed a longitudinal or repeated cross-sectional design, and quantitatively assessed with clinical questionnaires the effect of the COVID-19 pandemic or a related stressor on mental health indicators in community samples of children and adolescents.The results of our qualitative analysis of 69 studies indicate a general trend of less psychological well-being and more mental health problems, such as heightened stress, and depressive and anxiety symptoms during the pandemic. Data suggest that both protection measure intensity and infection dynamics were positively associated with severity of the psychopathology. The most reported influencing factors were age, gender, socio-economic status, previous state of mental and physical health, self-regulation abilities, parental mental health, parenting quality, family functioning, social support, isolation and loneliness, health-related worries, and consistent routines and structure. Our results demonstrate that children and adolescents worldwide have experienced more mental health problems due to the COVID-19 pandemic. They call for improved access to child and adolescent mental health care and prioritisation of child and adolescent welfare in political decision making.

Reliable cell retention of mammalian suspension cells in microfluidic cultivation chambers.

Microfluidic cultivation, with its high level of environmental control and spatio-temporal resolution of cellular behavior, is a well-established tool in today's microfluidics. Yet, reliable retention of (randomly) motile cells inside designated cultivation compartments still represents a limitation, which prohibits systematic single-cell growth studies. To overcome this obstacle, current approaches rely on complex multilayer chips or on-chip valves, which makes their application for a broad community of users infeasible. Here, we present an easy-to-implement cell retention concept to withhold cells inside microfluidic cultivation chambers. By introducing a blocking structure into a cultivation chamber's entrance and nearly closing it, cells can be manually pushed into the chamber during loading procedures but are unable to leave it autonomously in subsequent long-term cultivation. CFD simulations as well as trace substance experiments confirm sufficient nutrient supply within the chamber. Through preventing recurring cell loss, growth data obtained from Chinese hamster ovary cultivation on colony level perfectly match data determined from single-cell data, which eventually allows reliable high throughput studies of single-cell growth. Due to its transferability to other chamber-based approaches, we strongly believe that our concept is also applicable for a broad range of cellular taxis studies or analyses of directed migration in basic or biomedical research.

Analyzing Person-Place Interactions During Walking Episodes: Innovative Ambulatory Assessment Approach of Walking-Triggered e-Diaries.

BACKGROUND: Walking behavior is positively associated with physiological and mental health as much evidence has already shown. Walking is also becoming a critical issue for health promotion in urban environments as it is the most often used form of active mobility and helps to replace carbon dioxide emissions from motorized forms of transport. It therefore contributes to mitigate the negative effects of climate change and heat islands within cities. However, to promote walking among urban dwellers and to utilize its health-enhancing potential, we need to know more about the way in which physical and social environments shape individual experiences during walking episodes. Such person-place interactions could not adequately be analyzed in former studies owing to methodological constraints. OBJECTIVE: This study introduces walking-triggered e-diaries as an innovative ambulatory assessment approach for time-varying associations, and investigates its accuracy with 2 different validation strategies. METHODS: The walking trigger consists of a combination of movement acceleration via an accelerometer and mobile positioning of the cellphone via GPS and transmission towers to track walking activities. The trigger starts an e-diary whenever a movement acceleration exceeds a predetermined threshold and participants' locations are identified as nonstationary outside a predefined place of residence. Every 420 (±300) seconds, repeated e-diaries were prompted as long as the trigger conditions were met. Data were assessed on 10 consecutive days. First, to investigate accuracy, we reconstructed walking routes and calculated a percentage score for all triggered prompts in relation to all walking routes where a prompt could have been triggered. Then, to provide data about its specificity, we used momentary self-reports and objectively assessed movement behavior to describe activity levels before the trigger prompted an e-diary. RESULTS: Data of 67 participants could be analyzed and the walking trigger led to 3283 e-diary prompts, from which 2258 (68.8%) were answered. Regarding accuracy, the walking trigger prompted an e-diary on 732 of 842 (86.9%) reconstructed walking routes. Further, in 838 of 1206 (69.5%) triggered e-diaries, participants self-reported that they were currently walking outdoors. Steps and acceleration movement was higher during these self-reported walking episodes than when participants denied walking outdoors (steps: 106 vs 32; acceleration>0.2 g in 58.4% vs 19% of these situations). CONCLUSIONS: Accuracy analysis revealed that walking-triggered e-diaries are suitable to collect different data of individuals' current experiences in situations in which a person walks outdoors. Combined with environmental data, such an approach increases knowledge about person-place interactions and provides the possibility to gain knowledge about user preferences for health-enhancing urban environments. From a methodological viewpoint, however, specificity analysis showed how changes in trigger conditions (eg, increasing the threshold for movement acceleration) lead to changes in accuracy.

How do people feel while walking in the city? Using walking-triggered e-diaries to investigate the association of social interaction and environmental greenness during everyday life walking.

Background: Light to moderate physical activity, which includes walking, is associated with positive effects on physical and mental health. However, concerning mental health, social and physical environmental factors are likely to play an important role in this association. This study investigates person-place interactions between environmental characteristics (greenness, social interaction) and momentary affective states during walking episodes. A within-subject design is implemented, in which affective states and environmental characteristics are assessed while participants are walking outside. Methods: On smartphones, coupled with a motion sensor (move3), e-diaries were triggered as soon as people walked 100 m outside. E-diaries assessed momentary affective states (valence, calmness, energetic arousal), and social interaction (walking alone; seeing other people while walking; interacting with other people; walking with a known person) between 6 am and 10 pm over nine days. The percentage of greenness was determined afterward from recorded GPS and GIS data. Demographics were collected in advance via an online questionnaire. Multilevel models were calculated with R for 46 individuals (age = 41.2, ± 13.2; 52% female). Results: Affective state dimension energetic arousal showed a significant association with social interaction and greenness, i.e., participants rated energetic arousal lower when walking alone, and if there was less greenness vs. when interacting shortly with someone while walking (ß = 0.13, p = 0.02), and being in situations with more greenness (ß = 0.08, p = 0.02). Furthermore, associations with social interaction and greenness were found for dimension calmness: walking together with someone was associated with higher calmness (ß = 0.16, p = 0.02), and the higher the proportion of surrounding greenness during a walk, the higher calmness was rated, i.e., participants were calmer (ß = 0.09, p = 0.01). Significant associations with valence were not present. Conclusion: The findings indicate that the affective states varied significantly due to different social and physical environmental factors. In the future, the importance of environmental factors should be further investigated, e.g., by assessing environmental factors right in situations contrary to a subsequent imputation. Within-subject designs, and in particular triggered assessments with the addition of GPS, can aid in developing interventions for health-promoting urban environments.

Single-Cell Analysis of CHO Cells Reveals Clonal Heterogeneity in Hyperosmolality-Induced Stress Response.

Hyperosmolality can occur during industrial fed-batch cultivation processes of Chinese hamster ovary (CHO) cells as highly concentrated feed and base solutions are added to replenish nutrients and regulate pH values. Some effects of hyperosmolality, such as increased cell size and growth inhibition, have been elucidated by previous research, but the impact of hyperosmolality and the specific effects of the added osmotic-active reagents have rarely been disentangled. In this study, CHO cells were exposed to four osmotic conditions between 300 mOsm/kg (physiologic condition) and 530 mOsm/kg (extreme hyperosmolality) caused by the addition of either high-glucose-supplemented industrial feed or mannitol as an osmotic control. We present novel single-cell cultivation data revealing heterogeneity in mass gain and cell division in response to these treatments. Exposure to extreme mannitol-induced hyperosmolality and to high-glucose-oversupplemented feed causes cell cycle termination, mtDNA damage, and mitochondrial membrane depolarization, which hints at the onset of premature stress-induced senescence. Thus, this study shows that both mannitol-induced hyperosmolality (530 mOsm/kg) and glucose overfeeding induce severe negative effects on cell growth and mitochondrial activity; therefore, they need to be considered during process development for commercial production.

Assessing Visual Avoidance of Faces During Real-Life Social Stress in Children with Social Anxiety Disorder: A Mobile Eye-Tracking Study.

This study measured visual attention (fixation count, dwell time) during two real-life social stress tasks using mobile eye-tracking glasses in children (9-13 years) diagnosed with social anxiety disorder (SAD; n = 25) and a healthy control group (HC; n = 30). The influence of state anxiety on attention allocation and negative self-evaluation biases regarding gaze behavior were also examined. Compared to the HC group, children with SAD showed visual avoidance (i.e., fewer fixations) of the faces of interaction partners during the second social stress task. While visual avoidance in HC children decreased with declining state anxiety from the first to the second social stress task, no such effect was found in children with SAD. A negative self-evaluation bias regarding gaze behavior in children with SAD was not found. In sum, measuring visual attention during real-life social situations may help enhance our understanding of social attention in childhood SAD.

Genetically Encoded Ratiometric pH Sensors for the Measurement of Intra- and Extracellular pH and Internalization Rates.

pH-sensitive fluorescent proteins as genetically encoded pH sensors are promising tools for monitoring intra- and extracellular pH. However, there is a lack of ratiometric pH sensors, which offer a good dynamic range and can be purified and applied extracellularly to investigate uptake. In our study, the bright fluorescent protein CoGFP_V0 was C-terminally fused to the ligand epidermal growth factor (EGF) and retained its dual-excitation and dual-emission properties as a purified protein. The tandem fluorescent variants EGF-CoGFP-mTagBFP2 (pK' = 6.6) and EGF-CoGFP-mCRISPRed (pK' = 6.1) revealed high dynamic ranges between pH 4.0 and 7.5. Using live-cell fluorescence microscopy, both pH sensor molecules permitted the conversion of fluorescence intensity ratios to detailed intracellular pH maps, which revealed pH gradients within endocytic vesicles. Additionally, extracellular binding of the pH sensors to cells expressing the EGF receptor (EGFR) enabled the tracking of pH shifts inside cultivation chambers of a microfluidic device. Furthermore, the dual-emission properties of EGF-CoGFP-mCRISPRed upon 488 nm excitation make this pH sensor a valuable tool for ratiometric flow cytometry. This high-throughput method allowed for the determination of internalization rates, which represents a promising kinetic parameter for the in vitro characterization of protein-drug conjugates in cancer therapy.

Development and Psychometric Properties of a Computer-Based Standardized Emotional Competence Inventory (MeKKi) for Preschoolers and School-Aged Children.

A computer-based emotional competence inventory for preschoolers and school-aged children (MeKKi) was developed to assess five components of emotional competence: emotion vocabulary, emotion identification (situational, visual, auditory), emotion understanding, emotion expression, and emotion regulation. Validity, reliability, and factor structure were examined in a community sample of 313 preschoolers and school-aged children (164 boys, 145 girls, 4 n.a.) age 4-11 years (M = 6.35 years, SD = 1.85). Item statistics and Cronbach's α were calculated for the subscales. The unidimensionality of the subscales was additionally tested via item response theory or confirmatory factor analysis. Internal consistency (α) was overall satisfactory at 0.82, though the consistencies of the Visual and Auditory Emotion Identification subscales were lower. Unidimensionality was demonstrated for all subscales except Emotion Understanding. Results provide support for the use of the MeKKi in research and clinical settings to assess emotional competence.

Effects of Cognitive Reappraisal on Subjective and Neural Reactivity to Angry Faces in Children with Social Anxiety Disorder, Clinical Controls with Mixed Anxiety Disorders and Healthy Children.

Cognitive models of social anxiety suggest that social anxiety disorder (SAD) is characterized by both enhanced emotional reactivity and deficits in emotion regulation. Emotional reactivity to socially threatening children's faces and their modulation through reappraisal were measured via subjective ratings and electrocortical responses in children (age 10-13) with SAD (n = 28), clinical controls with mixed anxiety disorders (n = 28), and healthy controls (n = 29). Children with SAD showed higher subjective reactivity to the images of angry children's faces while all children reported reduced reactivity in their subjective ratings following reappraisal. Reduced electrocortical reactivity after reappraisal was only evident in older children and boys and was unrelated to anxiety. The present study indicates that cognitive reappraisal may be beneficial in reducing subjective reactivity in children with anxiety disorders, while neural effects of reappraisal may emerge at older ages.

Are you looking or looking away? Visual exploration and avoidance of disgust- and fear-stimuli: An eye-tracking study.

The present eye-tracking study investigates whether individuals with different levels of contamination fear are more likely to focus their attention on disgusting stimuli to explore them or to pull their attention away to avoid them. Ninety-two nonclinical participants with varying degrees of contamination fear performed a dot probe task. Eye-tracking methodology was employed to record gaze patterns during the task. In each trial participants looked at two pictures (disgust-neutral, fear-neutral, or neutral-neutral). Participants were further asked about the contents of the picture and the confidence of their answer. The results show that reaction times were slower and fixations were longer and more frequent for disgust targets compared with fear and neutral targets, which was further amplified in individuals with high contamination fear. However, image details of disgusting pictures were remembered less correctly than details of fearful pictures. These findings confirm the idea that attention on disgusting stimuli is maintained. Further, the findings indicate that disgust is processed more superficially and might also be related to disgust-associated uncertainty and ambiguity. The results may help to understand the difficult-to-change nature of disgust, which could be relevant to habituation and extinction processes in exposure therapy. (PsycInfo Database Record (c) 2022 APA, all rights reserved).

How to Perform a Microfluidic Cultivation Experiment-A Guideline to Success.

As a result of the steadily ongoing development of microfluidic cultivation (MC) devices, a plethora of setups is used in biological laboratories for the cultivation and analysis of different organisms. Because of their biocompatibility and ease of fabrication, polydimethylsiloxane (PDMS)-glass-based devices are most prominent. Especially the successful and reproducible cultivation of cells in microfluidic systems, ranging from bacteria over algae and fungi to mammalians, is a fundamental step for further quantitative biological analysis. In combination with live-cell imaging, MC devices allow the cultivation of small cell clusters (or even single cells) under defined environmental conditions and with high spatio-temporal resolution. Yet, most setups in use are custom made and only few standardised setups are available, making trouble-free application and inter-laboratory transfer tricky. Therefore, we provide a guideline to overcome the most frequently occurring challenges during a MC experiment to allow untrained users to learn the application of continuous-flow-based MC devices. By giving a concise overview of the respective workflow, we give the reader a general understanding of the whole procedure and its most common pitfalls. Additionally, we complement the listing of challenges with solutions to overcome these hurdles. On selected case studies, covering successful and reproducible growth of cells in MC devices, we demonstrate detailed solutions to solve occurring challenges as a blueprint for further troubleshooting. Since developer and end-user of MC devices are often different persons, we believe that our guideline will help to enhance a broader applicability of MC in the field of life science and eventually promote the ongoing advancement of MC.

Growth and eGFP Production of CHO-K1 Suspension Cells Cultivated From Single Cell to Laboratory Scale.

Scaling down bioproduction processes has become a major driving force for more accelerated and efficient process development over the last decades. Especially expensive and time-consuming processes like the production of biopharmaceuticals with mammalian cell lines benefit clearly from miniaturization, due to higher parallelization and increased insights while at the same time decreasing experimental time and costs. Lately, novel microfluidic methods have been developed, especially microfluidic single-cell cultivation (MSCC) devices have been proved to be valuable to miniaturize the cultivation of mammalian cells. So far, growth characteristics of microfluidic cultivated cell lines were not systematically compared to larger cultivation scales; however, validation of a miniaturization tool against initial cultivation scales is mandatory to prove its applicability for bioprocess development. Here, we systematically investigate growth, morphology, and eGFP production of CHO-K1 cells in different cultivation scales ranging from a microfluidic chip (230 nl) to a shake flask (125 ml) and laboratory-scale stirred tank bioreactor (2.0 L). Our study shows a high comparability regarding specific growth rates, cellular diameters, and eGFP production, which proves the feasibility of MSCC as a miniaturized cultivation tool for mammalian cell culture. In addition, we demonstrate that MSCC provides insights into cellular heterogeneity and single-cell dynamics concerning growth and production behavior which, when occurring in bioproduction processes, might severely affect process robustness.

Human Blood Serum Induces p38-MAPK- and Hsp27-Dependent Migration Dynamics of Adult Human Cardiac Stem Cells: Single-Cell Analysis via a Microfluidic-Based Cultivation Platform.

Migratory capabilities of adult human stem cells are vital for assuring endogenous tissue regeneration and stem cell-based clinical applications. Although human blood serum has been shown to be beneficial for cell migration and proliferation, little is known about its impact on the migratory behavior of cardiac stem cells and underlying signaling pathways. Within this study, we investigated the effects of human blood serum on primary human cardiac stem cells (hCSCs) from the adult heart auricle. On a technical level, we took advantage of a microfluidic cultivation platform, which allowed us to characterize cell morphologies and track migration of single hCSCs via live cell imaging over a period of up to 48 h. Our findings showed a significantly increased migration distance and speed of hCSCs after treatment with human serum compared to control. Exposure of blood serum-stimulated hCSCs to the p38 mitogen-activated protein kinase (p38-MAPK) inhibitor SB239063 resulted in significantly decreased migration. Moreover, we revealed increased phosphorylation of heat shock protein 27 (Hsp27) upon serum treatment, which was diminished by p38-MAPK-inhibition. In summary, we demonstrate human blood serum as a strong inducer of adult human cardiac stem cell migration dependent on p38-MAPK/Hsp27-signalling. Our findings further emphasize the great potential of microfluidic cultivation devices for assessing spatio-temporal migration dynamics of adult human stem cells on a single-cell level.

Rumination in Children with Social Anxiety Disorder: Effects of Cognitive Distraction and Relation to Social Stress Processing.

According to cognitive models of social anxiety disorder (SAD), both anticipatory processing and post-event processing are core mechanisms in disorder maintenance leading to dysfunctional coping with social situations through negative self-evaluation and increased anxiety. To date, little is known about these processes during late childhood, a critical period for disorder development. Further, it remains unclear if dysfunctional rumination in children can be altered through psychotherapeutic interventions such as cognitive distraction. In the current study, children aged 9 to 13 years with SAD and age- and gender-matched healthy controls (HCs, each: n = 30) participated in an experimental laboratory social stress task while anticipatory processing, post-event processing, subjective anxiety, self-evaluations, and autonomic arousal (skin conductance level) were assessed. Further, the impact of a brief cognitive distraction intervention on post-event processing was assessed. Children with SAD reported more negative anticipatory and post-event processing compared to HC children. Further, negative anticipatory processing was associated with higher subjective anxiety and reduced subjective performance ratings during the social stress task. In the aftermath of the stressor, distraction led to reduced subjective anxiety in the group with SAD and lower autonomic arousal in all children but did not alter post-event processing. The current study suggests that both anticipatory and post-event processing already play a key role in the maintenance of SAD in childhood. While distraction may be beneficial in reducing prolonged subjective anxiety and autonomic arousal after social situations, more research on interventions targeting ruminative processes is needed.

Towards an automatic analysis of CHO-K1 suspension growth in microfluidic single-cell cultivation.

MOTIVATION: Innovative microfluidic systems carry the promise to greatly facilitate spatio-temporal analysis of single cells under well-defined environmental conditions, allowing novel insights into population heterogeneity and opening new opportunities for fundamental and applied biotechnology. Microfluidics experiments, however, are accompanied by vast amounts of data, such as time series of microscopic images, for which manual evaluation is infeasible due to the sheer number of samples. While classical image processing technologies do not lead to satisfactory results in this domain, modern deep-learning technologies, such as convolutional networks can be sufficiently versatile for diverse tasks, including automatic cell counting as well as the extraction of critical parameters, such as growth rate. However, for successful training, current supervised deep learning requires label information, such as the number or positions of cells for each image in a series; obtaining these annotations is very costly in this setting. RESULTS: We propose a novel machine-learning architecture together with a specialized training procedure, which allows us to infuse a deep neural network with human-powered abstraction on the level of data, leading to a high-performing regression model that requires only a very small amount of labeled data. Specifically, we train a generative model simultaneously on natural and synthetic data, so that it learns a shared representation, from which a target variable, such as the cell count, can be reliably estimated. AVAILABILITY AND IMPLEMENTATION: The project is cross-platform, open-source and free (MIT licensed) software. We make the source code available at https://github.com/dstallmann/cell_cultivation_analysis; the dataset is available at https://pub.uni-bielefeld.de/record/2945513.

Development and application of a cultivation platform for mammalian suspension cell lines with single-cell resolution.

In bioproduction processes, cellular heterogeneity can cause unpredictable process outcomes or even provoke process failure. Still, cellular heterogeneity is not examined systematically in bioprocess research and development. One reason for this shortcoming is the applied average bulk analyses, which are not able to detect cell-to-cell differences. In this study, we present a microfluidic tool for mammalian single-cell cultivation (MaSC) of suspension cells. The design of our platform allows cultivation in highly controllable environments. As a model system, Chinese hamster ovary cells (CHO-K1) were cultivated over 150 h. Growth behavior was analyzed on a single-cell level and resulted in growth rates between 0.85 and 1.16 day-1 . At the same time, heterogeneous growth and division behavior, for example, unequal division time, as well as rare cellular events like polynucleation or reversed mitosis were observed, which would have remained undetected in a standard population analysis based on average measurements. Therefore, MaSC will open the door for systematic single-cell analysis of mammalian suspension cells. Possible fields of application represent basic research topics like cell-to-cell heterogeneity, clonal stability, pharmaceutical drug screening, and stem cell research, as well as bioprocess related topics such as media development and novel scale-down approaches.

The role of emotion regulation in socially anxious children and adolescents: a systematic review.

While numerous studies suggest that emotion dysregulation is important in maintaining social anxiety among adults, the role of emotion regulation in children and adolescents with social anxiety is not yet well understood. In this systematic review, we use the process model of emotion regulation as a framework for understanding emotion regulation in children and adolescents with social anxiety. We performed a systematic literature search in the electronic data bases Medline and PsycINFO. Additional studies were identified by hand search. We identified 683 studies, screened their titles and abstracts, viewed 142 studies, and included 55 of these. Study results indicate that children and adolescents with social anxiety disorder or high social anxiety show emotion dysregulation across all five domains of emotion regulation, such as enhanced social avoidance, more safety behaviors, repetitive negative thinking, biased attention and interpretation of social information, and reduced emotional expression. While enhanced social avoidance seems to be specific to childhood social anxiety, other maladaptive emotion regulation strategies, such as repetitive negative thinking, seem to occur transdiagnostically across different childhood anxiety disorders. Implications for current theory, interventions and future research are discussed.

Behavioral observation of prosocial behavior and social initiative is related to preschoolers' psychopathological symptoms.

Current research on preschool-age psychopathology suggests specific impairments in the two domains of social competence-prosocial behavior and social initiative-in children with externalizing and internalizing symptoms. While behavioral observation methods have been largely neglected in the past, they may extend the predominating questionnaire-based assessment as they allow for a precise and objective assessment of children's social behavior. In this study, we aimed to investigate whether prosocial behavior and social initiative measured in a limited resource task are related to externalizing and internalizing symptoms in a preschool-age community sample (N = 117, M = 4.67 years, SD = 2.75 months, females = 57). Externalizing and internalizing symptoms were rated by teachers (n = 109) and parents (n = 77) using the Strengths and Difficulties Questionnaire and by children using the Berkeley Puppet Interview (n = 97). Reduced prosocial actions were related to children's higher ratings of externalizing symptoms, while reduced social initiative actions were related to parents' higher ratings of internalizing symptoms. Prosocial behavior in the behavioral task was a marginally significant positive predictor of internalizing symptoms from children's perspective. These results highlight the value of behavioral observation measures and contribute to our understanding of interpersonal deficits already related to psychopathology at preschool age.