Quantification of cytosolic plasmid DNA degradation using high‐throughput sequencing: implications for gene delivery

Background Although cytosolic DNA degradation plays an important role in decreasing transgene expression, the plasmid degradation pattern remains largely unexplored. Methods Illumina dye sequencing was employed to provide degradation site information for S1 and cytosolic nucleases. S1 nuclease provided a positive control for a comparison between the agarose gel method and sequencing approaches. Results The poly(A) region between the β‐lactamase gene and the cytomegalovirus (CMV) promoter was identified as the most likely cut site for polyplex‐treated cytosol. The second most likely site, at the 5' end of the β‐lactamase gene, was identified by gel electrophoresis and sequencing. Additional sites were detected in the OriC region, the SV40/poly(A) region, the luciferase gene and the CMV promoter. Sequence analysis of plasmid treated with cytosol from control cells showed the greatest cut activity in the OriC region, the β‐lactamase gene and the poly(A) region following the luciferase gene. Additional regions of cut activity include the SV40 promoter and the β‐lactamase poly(A) termination sequence. Both cytosolic nucleases and the S1 nuclease showed substantial activity at the bacterial origin of replication ( OriC ). Conclusions High‐throughput plasmid sequencing revealed regions of the luciferase plasmid DNA sequence that are sensitive to cytosolic nuclease degradation. This provides new targets for improving plasmid and/or polymer design to optimize the likelihood of protein expression. Copyright © 2014 John Wiley & Sons, Ltd.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/106947/1/jgm2761.pd

Crowding and Queuing in Entrance Scenarios: Influence of Corridor Width in Front of Bottlenecks

In this paper, we present results of an entrance experiment investigating the effect of the corridor width in front of a bottleneck on the density. The idea is based on a previous study suggesting that a guiding system in front of an entrance can reduce pushing of the waiting people and thus the density at the entrance. In our study we aim to find out to what extend the corridor width has an impact on crowding or queuing behavior and with that on the density. The results of the presented experiment suggest that the transition takes place between a corridor width of 1.2 m and 2.3 m. The total duration of each experimental run is not significantly influenced by the corridor width but by the width of the entrance itself, the number of participants and partly by the motivation. In general, the density in front of the gate as well as the area of high density is increased by widening the corridor and by intensifying the motivation of the participants. However, the results also suggest that also the number of participants significantly influences the occurrence of pushing and the level of density

Screening zur frühen Identifizierung des psychosozialen Unterstützungsbedarfs bei Kindern psychisch erkrankter Eltern

Background: Children of mentally ill parents have an increased risk for mental disorders themselves, which is increased once again by the experience of abuse and neglect. Objective: The aim of this study is to present the development and preliminary piloting of a specific screening procedure that enables an early identification of a possible need for support in children with mentally ill parents. Material and methods: For children of mentally ill parents, two screening procedures were developed. In the parent questionnaire of the "children screening" mental abnormalities of the children and adolescents (6-18 years old) as well as resources are collated in an abbreviated form (mental abnormalities: 26 items, available resources: 4 items). In the "family screening" familial risk and stress factors (8 items) are also enquired about in a self-report of the parents as well as indications for abuse and neglect in the family (9 items). Results: Both screening procedures are easy to evaluate with the help of a traffic light system. The responsible psychiatrist receives a preliminary estimation and definite indications on the current need for psychosocial treatment and support of the whole family and particularly the children. Subsequently, they can initiate further steps for the integrated care of the family. Conclusion: The children and family screening procedures are economical in time and easily understandable screening procedures for identification of the need for treatment and support of the family. Subsequently, after the final validation it is planned to systematically implement the screening procedures in the psychiatric care of mentally ill parents. In this way the far-reaching negative consequences for these children can be prevented

Almost 2 years into the COVID-19 pandemic: an update on parental stress, parent mental health, and the occurrence of child maltreatment

Increased parental stress, poorer mental health, and an increase in the occurrence of child maltreatment (CM) have been reported in earlier phases of the COVID-19 pandemic. However, data from later phases of the pandemic are not yet available. We conducted a cross-sectional, representative survey among 1087 parents (48.8% female; mean age 41.72 years, SD = 9.15) in Germany in December 2021. Data were compared to a previous representative sample, assessed in August 2020 (N = 1024), and to normative scores of the outcome measures. Predictors for the occurrence of CM were analyzed by logistic regression. Pandemic-related stress and general stress were higher and physical and mental health were poorer in the December 2021 sample than in the August 2020 sample. Occurrence rates of CM varied between 5 and 56%. Verbal emotional abuse (n = 607, 56%), witnessing domestic violence (n = 446, 41%), and emotional neglect (n = 435, 40%) were most frequently reported. For these subtypes, parental risk for alcohol abuse (OR 2.1–2.7) and parental recent experience of violence (OR 2.1–5.1) were the strongest predictors. Across all subtypes of CM, parents reporting child maltreatment showed poorer scores on all stress outcomes, with medium–large-effect sizes. Results confirm a high burden within the families, almost 2 years into the pandemic. Occurrence rates of a broad spectrum of CM subtypes raise further concerns for the well-being of children. Family-oriented intervention efforts are needed to stabilize families and provide targeted support. Longitudinal studies are needed for a description of families at risk for poorer outcomes

Probiotics and the microbiota-gut-brain axis: focus on psychiatry

Purpose of Review Probiotics are living bacteria, which when ingested in adequate amounts, confer health benefits. Gutmicrobes are suggested to play a role in many psychiatric disorders and could be a potential therapeutic target. Between the gut and the brain, there is a bi-directional communication pathway called the microbiota-gut-brain axis. The purpose of this review is to examine data from recent interventional studies focusing on probiotics and the gut-brain axis for the treatment of depression, anxiety and schizophrenia.Recent Findings Probiotics are likely to improve depression but not schizophrenia. Regarding anxiety, there is only one trial which showed an effect of a multispecies probiotic. However, determinants like the duration of treatment, dosage and interactions have not been thoroughly investigated and deserve more scientific attention.Summary Microbiome-based therapies such as probiotics could be cautiously recommended for depression to enhance beneficial bacteria in the gut and to improve mood through the gut-brain axis

Spatiotemporal association between COVID-19 incidence and type 1 diabetes incidence among children and adolescents: a register-based ecological study in Germany

ObjectiveStudies have shown an increased incidence of pediatric type 1 diabetes during the COVID-19 pandemic, but the detailed role of SARS-CoV-2 infection in the incidence increase in type 1 diabetes remains unclear. We investigated the spatiotemporal association of pediatric type 1 diabetes and COVID-19 incidence at the district level in Germany.MethodsFor the period from March 2020 to June 2022, nationwide data on incident type 1 diabetes among children and adolescents aged <20 years and daily documented COVID-19 infections in the total population were obtained from the German Diabetes Prospective Follow-up Registry and the Robert Koch Institute, respectively. Data were aggregated at district level and seven time periods related to COVID-19 pandemic waves. Spatiotemporal associations between indirectly standardized incidence rates of type 1 diabetes and COVID-19 were analyzed by Spearman correlation and Bayesian spatiotemporal conditional autoregressive Poisson models.ResultsStandardized incidence ratios of type 1 diabetes and COVID-19 in the pandemic period were not significantly correlated across districts and time periods. A doubling of the COVID-19 incidence rate was not associated with a significant increase in the incidence rate of type 1 diabetes (relative risk 1.006, 95% CI 0.987; 1.019).ConclusionOur findings based on data from the pandemic period indirectly indicate that a causal relationship between SARS-COV-2 infection and type 1 diabetes among children and adolescents is unlikely