Degree of differentiation impacts neurobiological signature and resistance to hypoxia of SH-SY5Y cells.

Objective.SH-SY5Y cells are valuable neuronalin vitromodels for studying patho-mechanisms and treatment targets in brain disorders due to their easy maintenance, rapid expansion, and low costs. However, the use of various degrees of differentiation hampers appreciation of results and may limit the translation of findings to neurons or the brain. Here, we studied the neurobiological signatures of SH-SY5Y cells in terms of morphology, expression of neuronal markers, and functionality at various degrees of differentiation, as well as their resistance to hypoxia. We compared these to neurons derived from human induced pluripotent stem cells (hiPSCs), a well-characterized neuronalin vitromodel.Approach.We cultured SH-SY5Y cells and neurons derived from hiPSCs on glass coverslips or micro-electrode arrays. We studied expression of mature neuronal markers, electrophysiological activity, and sensitivity to hypoxia at various degrees of differentiation (one day up to three weeks) in SH-SY5Y cells. We used hiPSC derived neurons as a reference.Main results.Undifferentiated and shortly differentiated SH-SY5Y cells lacked neuronal characteristics. Expression of neuronal markers and formation of synaptic puncta increased during differentiation. Longer differentiation was associated with lower resistance to hypoxia. At three weeks of differentiation, MAP2 expression and vulnerability to hypoxia were similar to hiPSC-derived neurons, while the number of synaptic puncta and detected events were significantly lower. Our results show that at least three weeks of differentiation are necessary to obtain neurobiological signatures that are comparable to those of hiPSC-derived neurons, as well as similar sensitivities to metabolic stress. Significance.This indicates that extended differentiation protocols should be used to study neuronal characteristics and to model brain disorders with SH-SY5Y cells. We provided insights that may offer the basis for the utilization of SH-SY5Y cells as a more relevant neuronal model in the study of brain disorders.

The relation between social anxiety and biased interpretations in adolescents with mild intellectual disabilities.

BACKGROUND: Cognitive theories of anxiety emphasize the importance of cognitive processes in the onset and maintenance of anxiety disorders. However, little is known about these processes in children and adolescents with Mild Intellectual Disabilities (MID). AIMS: The aim of this study was to investigate interpretation bias and its content-specificity in adolescents with MID who varied in their levels of social anxiety. METHOD AND PROCEDURES: In total, 631 adolescents from seven special secondary schools for MID filled in questionnaires to measure their levels of social anxiety. They also completed the Interpretation Recognition Task to measure how they interpret ambiguous situations. OUTCOMES AND RESULTS: Adolescents with higher self-reported levels of social anxiety interpreted ambiguous scenarios as more negative than adolescents with lower self-reported social anxiety. Furthermore, this negative interpretation was specific for social situations; social anxiety was only associated with ambiguous social anxiety-related scenarios, but not with other anxiety-related scenarios. CONCLUSIONS AND IMPLICATIONS: These findings support the hypothesis that socially anxious adolescents with MID display an interpretation bias that is specific for stimuli that are relevant for their own anxiety. This insight is useful for improving treatments for anxious adolescents with MID by targeting content-specific interpretation biases.

Imagine the bright side of life: A randomized controlled trial of two types of interpretation bias modification procedure targeting adolescent anxiety and depression.

INTRODUCTION: Anxiety and depression are highly prevalent during adolescence and characterized by negative interpretation biases. Cognitive bias modification of interpretations (CBM-I) may reduce such biases and improve emotional functioning. However, as findings have been mixed and the traditional scenario training is experienced as relatively boring, a picture-based type of training might be more engaging and effective. METHODS: The current study investigated short- and long-term effects (up to 6 months) and users' experience of two types of CBM-I procedure in adolescents with heightened symptoms of anxiety or depression (N = 119, aged 12-18 year). Participants were randomized to eight online sessions of text-based scenario training, picture-word imagery training, or neutral control training. RESULTS: No significant group differences were observed on primary or secondary emotional outcomes. A decrease in anxiety and depressive symptoms, and improvements in emotional resilience were observed, irrespective of condition. Scenario training marginally reduced negative interpretation bias on a closely matched assessment task, while no such effects were found on a different task, nor for the picture-word or control group. Subjective evaluations of all training paradigms were relatively negative and the imagery component appeared particularly difficult for adolescents with higher symptom levels. CONCLUSIONS: The current results question the preventive efficacy and feasibility of both CBM-I procedures as implemented here in adolescents.

Online visual search attentional bias modification for adolescents with heightened anxiety and depressive symptoms: A randomized controlled trial.

Anxiety and depression, which are highly prevalent in adolescence, are both characterized by a negative attentional bias. As Attentional Bias Modification (ABM) can reduce such a bias, and might also affect emotional reactivity, it could be a promising early intervention. However, a growing number of studies also report comparable improvements in both active and placebo groups. The current study investigated the effects of eight online sessions of visual search (VS) ABM compared to both a VS placebo-training and a no-training control group in adolescents with heightened symptoms of anxiety and/or depression (n = 108). Attention bias, interpretation bias, and stress-reactivity were assessed pre- and post-training. Primary outcomes of anxiety and depressive symptoms, and secondary measures of emotional resilience were assessed pre- and post-training and at three and six months follow-up. Results revealed that VS training reduced attentional bias compared to both control groups, with stronger effects for participants who completed more training sessions. Irrespective of training condition, an overall reduction in symptoms of anxiety and depression and an increase in emotional resilience were observed up to six months later. The training was evaluated relatively negatively. Results suggest that online ABM as employed in the current study has no added value as an early intervention in adolescents with heightened symptoms.

Emotional working memory training as an online intervention for adolescent anxiety and depression: A randomised controlled trial.

OBJECTIVE: Anxiety and depression are highly prevalent disorders in adolescence. They are associated with deficits in working memory (WM), which also appears to increase rumination, worry, and negative mood. WM training, especially in an emotional context, might help in reducing or preventing these disorders. The current study investigated the direct effects of online emotional WM training on WM capacity, and short- and long-term effects on symptoms of anxiety and depression, and secondary measures of emotional functioning. METHODS: Unselected adolescents (n = 168, aged 11-18) were randomised over an active or placebo emotional WM training. WM was assessed before and after 4 weeks of bi-weekly training. Emotional functioning was assessed pre- and post-training and at 3, 6, and 12 months follow-up. RESULTS: Improvements in WM capacity and both short- and long-term emotional functioning were found in both training groups, with the only group difference being a trend for a larger increase in self-esteem in the active group compared to the placebo group. CONCLUSIONS: The general improvements irrespective of training condition suggest non-specific training or time effects, or some shared active ingredient in both conditions. Future research is necessary to detect potentially effective components of (emotional) WM training and to increase adolescent engagement with online training.

Online attentional bias modification training targeting anxiety and depression in unselected adolescents: Short- and long-term effects of a randomized controlled trial.

Based on information processing models of anxiety and depression, we investigated the efficacy of multiple sessions of online attentional bias modification training to reduce attentional bias and symptoms of anxiety and depression, and to increase emotional resilience in youth. Unselected adolescents (N = 340, age: 11-18 years) were randomly allocated to eight sessions of a dot-probe, or a visual search-based attentional training, or one of two corresponding placebo control conditions. Cognitive and emotional measures were assessed pre- and post-training; emotional outcome measures also at three, six and twelve months follow-up. Only visual search training enhanced attention for positive information, and this effect was stronger for participants who completed more training sessions. Symptoms of anxiety and depression reduced, whereas emotional resilience improved. However, these effects were not especially pronounced in the active conditions. Thus, this large-scale randomized controlled study provided no support for the efficacy of the current online attentional bias modification training as a preventive intervention to reduce symptoms of anxiety or depression or to increase emotional resilience in unselected adolescents. However, the absence of biased attention related to symptomatology at baseline, and the large drop-out rates at follow-up preclude strong conclusions.

Seasonal changes in prescribing of long-acting beta-2-agonists-containing drugs.

BACKGROUND: For patients with asthma, COPD, or asthma-COPD overlap syndrome (ACOS), inter-country comparisons of seasonal changes in drug prescriptions are scarce or missing. Hence, we aimed to compare seasonal changes in prescription rates of long-acting beta-2-agonist (LABA) in four European countries. METHODS: A common study protocol was applied to six health care databases (Germany, Spain, the Netherlands (2), and the UK (2)) to calculate age- and sex-standardized point prevalence rates (PPRs) of LABA-containing prescriptions by the 1st of March, June, September, and December of each year during the study period 2002-2009. Seasonal variation of PPRs was quantified using seasonal indexes (SIs; based on the ratio-to-moving-average-method) and SIs averaged over the study period (aSI) stratified by sex, age, and indication (asthma, COPD, or ACOS). RESULTS: There was a moderate seasonal change in LABA-containing prescriptions which was more pronounced in asthma or COPD patients compared to ACOS patients. For asthma and ACOS patients, highest seasonal variation was found for patients living in Spain (aSI: 87.3-110.7, aSI: 93.2-103.1) whereas for COPD highest seasonal variation was revealed for the NPCRD database (the Netherlands) (aSI: 92.2-105.6). Regarding age and sex, highest seasonal variation was found in Spanish boys under 10 years of age having a diagnosis of asthma. CONCLUSIONS: By applying a common analysis in six databases, we could observe moderate overall seasonal changes in LABA-containing prescription rates in patients with asthma, COPD, or ACOS.

Amygdala activation during emotional face processing in adolescents with affective disorders: the role of underlying depression and anxiety symptoms.

Depressive and anxiety disorders are often first diagnosed during adolescence and it is known that they persist into adulthood. Previous studies often tried to dissociate depressive and anxiety disorders, but high comorbidity makes this difficult and maybe even impossible. The goal of this study was to use neuroimaging to test what the unique contribution is of depression and anxiety symptomatology on emotional processing and amygdala activation, and to compare the results with a healthy control group. We included 25 adolescents with depressive and/or anxiety disorders and 26 healthy adolescents. Participants performed an emotional face processing task while in the MRI scanner. We were particularly interested in the relation between depression/anxiety symptomatology and patterns of amygdala activation. There were no significant differences in activation patterns between the control group and the clinical group on whole brain level and ROI level. However, we found that dimensional scores on an anxiety but not a depression subscale significantly predicted brain activation in the right amygdala when processing fearful, happy and neutral faces. These results suggest that anxiety symptoms are a better predictor for differentiating activation patterns in the amygdala than depression symptoms. Although the current study includes a relatively large sample of treatment naïve adolescents with depression/anxiety disorders, results might be influenced by differences between studies in recruitment strategies or methodology. Future research should include larger samples with a more equal distribution of adolescents with a clinical diagnosis of depression and/or anxiety. To conclude, this study shows that abnormal amygdala responses to emotional faces in depression and anxiety seems to be more dependent on anxiety symptoms than on depression symptoms, and thereby highlights the need for more research to better characterize clinical groups in future studies.

Visual search attentional bias modification reduced social phobia in adolescents.

BACKGROUND AND OBJECTIVES: An attentional bias for negative information plays an important role in the development and maintenance of (social) anxiety and depression, which are highly prevalent in adolescence. Attention Bias Modification (ABM) might be an interesting tool in the prevention of emotional disorders. The current study investigated whether visual search ABM might affect attentional bias and emotional functioning in adolescents. METHODS: A visual search task was used as a training paradigm; participants (n = 16 adolescents, aged 13-16) had to repeatedly identify the only smiling face in a 4 × 4 matrix of negative emotional faces, while participants in the control condition (n = 16) were randomly allocated to one of three placebo training versions. An assessment version of the task was developed to directly test whether attentional bias changed due to the training. Self-reported anxiety and depressive symptoms and self-esteem were measured pre- and post-training. RESULTS: After two sessions of training, the ABM group showed a significant decrease in attentional bias for negative information and self-reported social phobia, while the control group did not. There were no effects of training on depressive mood or self-esteem. LIMITATIONS: No correlation between attentional bias and social phobia was found, which raises questions about the validity of the attentional bias assessment task. Also, the small sample size precludes strong conclusions. CONCLUSIONS: Visual search ABM might be beneficial in changing attentional bias and social phobia in adolescents, but further research with larger sample sizes and longer follow-up is needed.

Head-only followed by cardiac arrest electrical stunning is an effective alternative to head-only electrical stunning in pigs.

Many small slaughter facilities use head-only electrical stunning to render swine unconscious and insensible to pain before slaughter. Head-only electrical stunning is a reversible procedure that is optimally effective for approximately 15 s after stun completion. In many small North American slaughter plants, the authors have observed hoist speeds that are too slow to achieve a short enough stun-to-bleed interval to maintain insensibility through exsanguination. Unlike many European plants, there is no separate high-speed hoist for pigs and exsanguination on the floor is not condoned. As a result, a 2-stage stunning method was proposed where head-only stunning for 3 s was immediately followed by application of the same stunning wand to the cardiac region of the animal for 3 s while lying in lateral recumbancy. A paired-comparison study was conducted on 89 pigs in a small slaughter facility to compare the head-only method applied for 6 s with the head/heart method. The objective was to evaluate signs of return to sensibility, stun-to-bleed time, blood lactate concentration, muscle pH, drip loss, and fresh meat color to validate the head/heart electrical stunning method for small slaughter plants. Incidence of corneal reflex was not different (P > 0.05) between head/heart (93.8%) and head only (85%) stunning. Nose twitching was more common (P < 0.05) in head only (26.5%) than head/heart (5%) stunning. Head/heart stunning eliminated rhythmic breathing, natural blinking, eye tracking to moving objects, and righting reflex, which were all observed in head-only stunned pigs. Eye tracking to moving objects was observed in 40.8% of head-only stunned pigs. Blood lactate was not different (P > 0.05) between stunning methods (head only: 8.8 ± 0.7 mmol/L, head/heart: 7.8 ± 0.7 mmol/L). Stun-to-bleed time did not differ (P > 0.05; head only: 32 ± 1 s, head/heart: 33 ± 1 s). Mean time to loss of heartbeat with the head-only method was 121 ± 5 s. No heartbeat was observed with the head/heart method. Longissimus thoracis pH, color, and drip loss were not different (P > 0.05) between stunning methods. This study determined that the head/heart electrical stunning method reduces the incidence of signs of return to sensibility without significant effects on meat quality, plant operation speed, or blood lactate concentration. In addition, the head/heart method requires no capital investment for plants that are currently using the head-only method.

Inhibition of trehalase activity enhances trehalose accumulation in transgenic plants.

As a first step toward the exploitation of the disaccharide trehalose as a stress-protective and preservative agent in plants, we engineered trehalose biosynthesis in tobacco (Nicotiana tabacum) and potato (Solanum tuberosum) by introducing the otsA and otsB genes from Escherichia coli, which encode trehalose-6-phosphate synthase and trehalose-6-phosphate phosphatase, respectively. In leaves of transgenic tobacco plants, very low levels of trehalose accumulation were obtained (0.11 mg g-1 fresh weight), whereas in transgenic potato tubers, no trehalose accumulated at all. Plant trehalase activity was shown to affect the accumulation of trehalose in these plants. An increase in trehalose accumulation, up to 0.41 and 4.04 mg g-1 fresh weight in tobacco leaves and potato micro-tubers, respectively, was noted when the potent trehalase inhibitor validamycin A was added to in vitro plants and to hydroponically grown greenhouse plants. Stunted growth and the formation of lancet-shaped leaves by trehalose-accumulating tobacco plants suggest a negative effect of trehalose biosynthesis on N. tabacum development. It is surprising that experiments with wild-type plants cultured in the presence of validamycin A indicate that, despite current belief, the capacity to synthesize trehalose may not be restricted to primitive phyla of vascular plants and certain "resurrection plants," but may exist throughout the angiosperms.