ABSTRACT
Transcription factor (TF)-promoter pairs have been repurposed from native hosts to provide tools to measure intracellular biochemical production titer and dynamically control gene expression. Most often, native TF-promoter systems require rigorous screening to obtain desirable characteristics optimized for biotechnological applications. High-throughput techniques may provide a rational and less labor-intensive strategy to engineer user-defined TF-promoter pairs using fluorescence-activated cell sorting and deep sequencing methods (sort-seq). Based on the designed promoter library's distribution characteristics, we elucidate sequence-function interactions between the TF and DNA. In this work, we use the sort-seq method to study the sequence-function relationship of a σ54-dependent, butanol-responsive TF-promoter pair, BmoR-PBMO derived from Thauera butanivorans, at the nucleotide level to improve biosensor characteristics, specifically an improved dynamic range. Activities of promoters from a mutagenized PBMO library were sorted based on gfp expression and subsequently deep sequenced to correlate site-specific sequences with changes in dynamic range. We identified site-specific mutations that increase the sensor output. Double mutant and a single mutant, CA(129,130)TC and G(205)A, in PBMO promoter increased dynamic ranges of 4-fold and 1.65-fold compared with the native system, respectively. In addition, sort-seq identified essential sites required for the proper function of the σ54-dependent promoter biosensor in the context of the host. This work can enable high-throughput screening methods for strain development.
ABSTRACT
Meyer, Erdberg, and Shaffer (2007) pointed to significant variability across nonpatient samples of children and adolescents included in the Comprehensive System (CS; Exner, 2003) international project. Consequently, Meyer et al. stated that applying standard interpretive guidelines for children and adolescents would result in some inaccurate inferences about mental health and recommended inferring difficulties in a child's protocol in cases in which a score on a given variable deviates from the most extreme normative samples' mean. In this Comment, we explore this idea and provide the clinician with basic guidelines for interpreting young patients' protocols in which structural data are used jointly with developmental and contextual factors. We present these guidelines in relation to a case study.
Subject(s)
Mental Health , Personality Assessment/statistics & numerical data , Rorschach Test/statistics & numerical data , Adolescent , Age Factors , Child , Humans , Mental Disorders/diagnosis , Meta-Analysis as Topic , Psychometrics , Reference Values , Reproducibility of Results , Research Design/standards , Surveys and QuestionnairesSubject(s)
Affect , Anorexia Nervosa/therapy , Bulimia Nervosa/therapy , Dependency, Psychological , Psychoanalytic Therapy/methods , Adolescent , Anorexia Nervosa/psychology , Bulimia Nervosa/psychology , Defense Mechanisms , Female , Humans , Longitudinal Studies , Mind-Body Relations, Metaphysical , Outcome Assessment, Health Care , Reality TestingABSTRACT
The study examined changes in ego functions and mental distress among eating disorder (ED) adolescent inpatients following integrative treatment. Participants included 31 restricting anorexia nervosa and 25 bingeing/purging spectrum ED inpatients. Rorschach Comprehensive System (CS; Exner, 2003) and self-report measures were administered on admission and discharge. We found that improvement in ego functions, mental distress and ED symptomatology occurred in both ED groups at discharge. Nevertheless, the two ED subgroups exhibited different configurations of change as well as varying patterns of relations between changes in mental distress and ego functions and improvement in ED symptoms. These data further indicate that the restricting and bingeing/purging patients may require different treatment approaches.
Subject(s)
Complementary Therapies/statistics & numerical data , Ego , Feeding and Eating Disorders/psychology , Feeding and Eating Disorders/therapy , Inpatients/psychology , Stress, Psychological/therapy , Adolescent , Anorexia Nervosa/psychology , Anorexia Nervosa/therapy , Body Mass Index , Bulimia Nervosa/psychology , Bulimia Nervosa/therapy , Feeding and Eating Disorders/diagnosis , Female , Humans , Patient Admission , Patient Discharge , Personality Inventory , Psychiatric Status Rating Scales , Self Concept , Surveys and Questionnaires , Treatment OutcomeABSTRACT
Conceptualizing eating disorders (EDs) as forming a coping mechanism against facing psychic pain, in this study, we examined the contribution of weight stabilization and ED behaviors to psychological outcomes of affective distress, defensiveness, and contact with inner/external reality among 2 subgroups of adolescent inpatients with (a) "restricting type" anorexia (AN-R) and (b) bingeing/purging type EDs (B/P). We administered Rorschach Comprehensive System (CS; Exner, 2003) and self-report measures at acute admission and on discharge. Data indicated that in both subgroups, stabilization of weight and disordered eating behaviors was accompanied by reduced explicit (self-reported) affective distress, yet by elevated implicit affective distress (D, SumShd). Subgroups' different Rorschach change trends indicated different dynamic processes in defensive style and contact with inner/external reality.
