The sessile nature of plants necessitates a phenotypic plasticity that enables plants to respond to changes in environment throughout growth and development. Orchestration of molecular components at various levels in a spatial and temporal dimension is required. However, how signals are integrated to produce a specific response, e.g. how a simple hydrocarbon, the plant hormone ethylene, can cause a diverse set of morphological response, remains elusive. To better understand the integration of signals involved in generating these phenotypes, we characterized the transcriptional regulation of the ethylene response by identifying key protein-DNA interactions and transcriptional profiles in a temporal manner. The plant hormone ethylene regulates numerous growth and developmental processes in plants, including stem cell division, differential cell growth, stress response, response to pathogens, germination, senescence, fruit ripening and the triple response. The triple response, a decrease in cellular elongation, increase in radial swelling, and an exaggerated apical hook, has been the hallmark of the ethylene response in dark grown seedlings. To determine the molecular mechanisms of the ethylene response, we characterized the dynamic ethylene transcriptional response in etiolated seedlings. We identified targets of the master regulator of the ethylene signaling pathway, ETHYLENE INSENSITIVE3 (EIN3), using temporal chromatin immunoprecipitation sequencing (ChIP- Seq) and transcript sequencing (mRNA-Seq). First we identified the minimum complement of genes required for the ethylene transcriptional response in etiolated seedlings. Then we characterized targets of EIN3 based on their transcriptional state as well as their function in relation of the ethylene signaling pathway and response. We found a singular EIN3 binding pattern that increased upon ethylene treatment, which did not correspond to target gene transcription. Binding of EIN3 established feedforward transcriptional cascades, feedback circuitry of the ethylene signaling pathway, and interconnections between hormone response pathways at a multitude of levels, e.g. hormone biosynthesis, signal reception, signaling, and transcriptional response. Finally, we showed that mutants in a gene family targeted by EIN3 exhibit hormone response phenotypes in several developmental stages, thus demonstrating the integral role EIN3 plays in the orchestration of hormone crosstalk in plant growth and development

Depression during sensitive periods of social development may have consequences that extend well beyond mental health, and far into adulthood. This study compared the social functioning of adults with adolescent-onset depression (ages 10-20) to those with adult-onset depression (ages 21+). Participants were 3,360 adults (67.2% female; ages 42 ± 15) who had experienced major depression. Adult functional outcomes were marital status, divorce, number of children, years of education, employment status, household income, dependency on welfare, and obesity. Participants with depression during adolescence were less likely to get married, have children, and more likely to have lower household incomes. Depression during adolescence may be associated with broader functional outcomes that impact individuals and society, and may be mitigated through intervention and effective policy.

Background

An increasing number of mental health apps (MHapps) are being developed for youth. In addition, youth are high users of both technologies and MHapps. However, little is known about their perspectives on MHapps. MHapps might be particularly well suited to reach the youth underserved by traditional mental health resources, and incorporating their perspectives is especially critical to ensure such tools are useful to them.

Objective

The goal of this study was to develop and pilot a process for eliciting youth perspectives on MHapps in a structured and collaborative way. We also sought to generate learnings on the perspectives of Latinx youth on MHapps and their use in ways that might facilitate discovery, activation, or engagement in MHapps, especially in Latinx populations.

Methods

We created a series of focus groups consisting of 5 sessions. The groups introduced different categories of MHapps (cognitive behavioral therapy apps, mindfulness apps, and miscellaneous apps). Within each category, we selected 4 MHapps that participants chose to use for a week and provided feedback through both between-session and in-session activities. We recruited 5 youths ranging in age from 15 to 21 (mean 18, SD 2.2) years. All the participants identified as Hispanic or Latinx. After completing all 5 focus groups, the participants completed a brief questionnaire to gather their impressions of the apps they had used.

Results

Our focus group methodology collected detailed and diverse information about youth perspectives on MHapps. However, we did identify some aspects of our methods that were less successful at engaging the youth, such as our between-session activities. The Latinx youth in our study wanted apps that were accessible, relatable, youth centric, and simple and could be integrated with their offline lives. We also found that the mindfulness apps were viewed most favorably but that the miscellaneous and cognitive behavioral therapy apps were viewed as more impactful.

Conclusions

Eliciting youth feedback on MHapps is critical if these apps are going to serve a role in supporting their mental health and well-being. We refined a process for collecting feedback from the youth and identified factors that were important to a set of Latinx youth. Future work could be broader, that is, recruit larger samples of more diverse youth, or deeper, that is, collect more information from each youth around interests, needs, barriers, or facilitators or better understand the various impacts of MHapps by using qualitative and quantitative measures. Nevertheless, this study advances the formative understanding of how the youth, particularly Latinx youth, might be viewing these tools.

Pluripotent stem cells offer great therapeutic promise for personalized treatment platforms for numerous injuries, disorders, and diseases. Octamer-binding transcription factor 4 (OCT4) is a key regulatory gene maintaining pluripotency and self-renewal of mammalian cells. With site-specific integration for gene correction in cellular therapeutics, use of the OCT4 promoter may have advantages when expressing a suicide gene if pluripotency remains. However, the human OCT4 promoter region is 4 kb in size, limiting the capacity of therapeutic genes and other regulatory components for viral vectors, and decreasing the efficiency of homologous recombination. The purpose of this investigation was to characterize the functionality of a novel 967bp OCT4-short response element during pluripotency and to examine the OCT4 titer-dependent response during differentiation to human derivatives not expressing OCT4. Our findings demonstrate that the OCT4-short response element is active in pluripotency and this activity is in high correlation with transgene expression in vitro, and the OCT4-short response element is inactivated when pluripotent cells differentiate. These studies demonstrate that this shortened OCT4 regulatory element is functional and may be useful as part of an optimized safety component in a site-specific gene transferring system that could be used as an efficient and clinically applicable safety platform for gene transfer in cellular therapeutics.

Abstract

BACKGROUND

Chemotherapy for high grade gliomas (HGG) is limited by the blood-brain-barrier (BBB). Convection enhanced delivery (CED) improves chemotherapy delivery by utilizing fluid convection obviating the challenges of crossing the BBB while minimizing systemic toxicity. CED of nanoliposomal-irinotecan (Onivyde) showed to be a superior delivery route for anti-tumor activity in animal models. An advance of this trial is the development and use of real time CED, which utilizes MRI to visualize the CED process with the aid of co-convected contrast agents, monitoring delivery into the brain and affording for corrective action.

METHODS

This is a 3 + 3 single dose escalation trial with 2 cohorts: 20mg/ml and 40mg/ml. Onivyde and GAD were co-infused via the same catheters in a one-time delivery. The total volume of infusate, and consequently total dose, were personalized based on the patient’s tumor volume, and ranged from 20–680 mg of Onivyde, given via up to 4 catheters. Tumor diameters were allowed to be 1 – 4 cm, with injection volumes ranging from 2 – 17 mL of infusate.

RESULTS

10 patients have been treated on this protocol, all in under 5 hours. There were 7 GBs, 2 anaplastic astrocytomas, and 1 oligoastrocytoma. Seven patients lived over a year after treatment, which is remarkable since median survival rates for multiply recurrent: GBs = 8 months, AAs = 11 months. Utilizing imaging software, we correlated pre-infusion modeling of the drug distribution with post-infusion imaging. A number of technical challenges were overcome by real time monitoring; the total volume of distribution (Vd), and the Vd to volume infused (Vi) ratio for each infusion was ~2.

CONCLUSIONS

Image-guided distribution allows for safe real-time placement and adjustment of CED cannula of Onivyde into patient’s brains. Such methods allow for maximum tumor coverage and warrant further studies with repeat dosing.

Urea cycle disorders are incurable enzymopathies that affect nitrogen metabolism and typically lead to hyperammonemia. Arginase deficiency results from a mutation in Arg1, the enzyme regulating the final step of ureagenesis and typically results in developmental disabilities, seizures, spastic diplegia, and sometimes death. Current medical treatments for urea cycle disorders are only marginally effective, and for proximal disorders, liver transplantation is effective but limited by graft availability. Advances in human induced pluripotent stem cell research has allowed for the genetic modification of stem cells for potential cellular replacement therapies. In this study, we demonstrate a universally-applicable CRISPR/Cas9-based strategy utilizing exon 1 of the hypoxanthine-guanine phosphoribosyltransferase locus to genetically modify and restore arginase activity, and thus ureagenesis, in genetically distinct patient-specific human induced pluripotent stem cells and hepatocyte-like derivatives. Successful strategies restoring gene function in patient-specific human induced pluripotent stem cells may advance applications of genetically modified cell therapy to treat urea cycle and other inborn errors of metabolism.

Background

Evaluation and interpretation of the literature on obstructive sleep apnea (OSA) allows for consolidation and determination of the key factors important for clinical management of the adult OSA patient. Toward this goal, an international collaborative of multidisciplinary experts in sleep apnea evaluation and treatment have produced the International Consensus statement on Obstructive Sleep Apnea (ICS:OSA).

Methods

Using previously defined methodology, focal topics in OSA were assigned as literature review (LR), evidence-based review (EBR), or evidence-based review with recommendations (EBR-R) formats. Each topic incorporated the available and relevant evidence which was summarized and graded on study quality. Each topic and section underwent iterative review and the ICS:OSA was created and reviewed by all authors for consensus.

Results

The ICS:OSA addresses OSA syndrome definitions, pathophysiology, epidemiology, risk factors for disease, screening methods, diagnostic testing types, multiple treatment modalities, and effects of OSA treatment on multiple OSA-associated comorbidities. Specific focus on outcomes with positive airway pressure (PAP) and surgical treatments were evaluated.

Conclusion

This review of the literature consolidates the available knowledge and identifies the limitations of the current evidence on OSA. This effort aims to create a resource for OSA evidence-based practice and identify future research needs. Knowledge gaps and research opportunities include improving the metrics of OSA disease, determining the optimal OSA screening paradigms, developing strategies for PAP adherence and longitudinal care, enhancing selection of PAP alternatives and surgery, understanding health risk outcomes, and translating evidence into individualized approaches to therapy.