Dataset on Galanin Receptor 3 mutants that improve recombinant receptor expression and stability in an agonist and antagonist bound form.

Galanin Receptor 3 (GALR3) is a G-protein-coupled receptor with a widespread distribution in the brain and plays a role in a variety of physiologic processes including cognition/memory, sensory/pain processing, hormone secretion, and feeding behavior. Therefore, GALR3 is considered an attractive CNS drug target (Freimann et al., 2015) [1]. This dataset contains GALR3 point mutants that improve recombinant protein expression and thermal stability of the receptor contained in virus-like particles (VLPs) or obtained by detergent-purification of baculovirus-infected insect cells. The mutations listed can be grouped in those that improve the stability of the agonist-bound and the antagonist-bound form of the receptor. Protein characteristics in terms of protein expression and thermal stability were comparable between GPCR-VLP and GPCR overexpressing Sf9 cultures. The further analysis and detailed results of these mutants as well as their impact on biophysical assay development for drug discovery can be found in "Method for Rapid Optimization of Recombinant GPCR Protein Expression and Stability using Virus-Like Particles" (Ho et al., 2017) [2].

Method for rapid optimization of recombinant GPCR protein expression and stability using virus-like particles.

Recent innovative approaches to stabilize and crystallize GPCRs have resulted in an unprecedented breakthrough in GPCR crystal structures as well as application of the purified receptor protein in biophysical and biochemical ligand binding assays. However, the protein optimization process to enable these technologies is lengthy and requires iterative overexpression, solubilization, purification and functional analysis of tens to hundreds of protein variants. Here, we report a new and versatile method to screen in parallel hundreds of GPCR variants in HEK293 produced virus-like particles (VLPs) for protein yield, stability, functionality and ligand binding. This approach reduces the time and resources during GPCR construct optimization by eliminating lengthy protein solubilization and purification steps and by its adaptability to many binding assay formats (label or label-free detection). We exemplified the robustness of our VLP method by screening 210 GALR3-VLP variants in a radiometric agonist-based binding assay and a subset of 88 variants in a label-free antagonist-based assay. The resulting GALR3 agonist or antagonist stabilizing variants were then further used for recombinant protein expression in transfected insect cells. The final purified protein variants were successfully immobilized on a biosensor chip and used in a surface plasmon resonance binding assay.

Post-concussion symptoms in mild traumatic brain injury: findings from a paediatric outpatient clinic.

Purpose Mild traumatic brain injury (mTBI) is common among children and is associated with a range of symptomatology and clinical presentations. This study uses data from a paediatric outpatient TBI clinic to (1) investigate characteristics associated with more severe post-concussive symptoms and (2) examine differences in the proportion of individuals endorsing specific post-concussion symptoms based on group (e.g., sex, type of injury, and psychiatric history). Methods Data from the Children's Hospital of Richmond's TBI outpatient programme were analysed (N = 157). Results Gender and sports injury were associated with severity of symptoms. In addition, females endorsed a greater number of overall symptoms than males. A number of specific symptoms were found to be endorsed to a greater extent based on psychiatric history and type of injury; however, overall total number of symptoms endorsed did not differ based on these characteristics. Conclusions Findings from this study provide further evidence that mTBI affects a wide range of youth and that associated symptomatology can indeed be varied. Moreover, results revealed differences in endorsement of specific symptoms and symptom severity based on patient and injury characteristics which have implications for concussion assessment and treatment. Implications for Rehabilitation Symptoms following mild traumatic brain injury (mTBI) in children and adolescents can have varied presentation, ranging from minimal to severe. Females and those with non-sports-related injuries are more likely to endorse greater symptoms following concussion. Symptom evaluation is an essential component of the concussion assessment and treatment of paediatric patients following mTBI, and clinicians should be aware of patient characteristics associated with increased symptoms, especially when baseline symptom data are not available.

Traumatic brain injury: a comparison of inpatient functional outcomes between children and adults.

OBJECTIVES: To examine age-related differences in functional outcomes following traumatic brain injury. PARTICIPANTS AND PROCEDURE: Seventy-six patients admitted to a pediatric acute rehabilitation hospital were compared with 2548 adult patients in the National Institute on Disability and Rehabilitation Research-funded traumatic brain injury model systems national database. MAIN OUTCOME MEASURES: Functional Independence Measure totals during inpatient rehabilitation. RESULTS: Increasing age was significantly associated with improved outcome in children and with poorer outcome in adults. CONCLUSION: The relationship between age and functional outcome is different within different age groups (pediatric vs adult), and the effect of moderating variables differs by age group.

Comparison of endodontist versus generalist regarding preference for postendodontic use of cotton pellets in pulp chamber.

A degree of uncertainty exists about the use of cotton pellets in the pulp chamber after canal obturation. The purpose of this study was to poll practicing endodontists and generalists regarding their preference. Forty-six endodontists and an equal number of general dentists were polled in a mail survey regarding their preferences for the use of a cotton pellet in the pulp chamber after canal obturation. Eighty-seven percent of the endodontists and 54% of the generalists responded. Of the endodontists, 62.5% said they thought that the general dentist wanted a cotton pellet, contrasted to 80% of the generalists. The difference in proportions was not statistically significant.