CRISPR-Cas9 ribonucleoprotein-mediated co-editing and counterselection in the rice blast fungus

The rice blast fungus Magnaporthe oryzae is the most serious pathogen of cultivated rice and a significant threat to global food security. To accelerate targeted mutation and specific genome editing in this species, we have developed a rapid plasmid-free CRISPR-Cas9-based genome editing method. We show that stable expression of Cas9 is highly toxic to M. oryzae. However efficient gene editing can be achieved by transient introduction of purified Cas9 pre-complexed to RNA guides to form ribonucleoproteins (RNPs). When used in combination with oligonucleotide or PCR-generated donor DNAs, generation of strains with specific base pair edits, in-locus gene replacements, or multiple gene edits, is very rapid and straightforward. We demonstrate a co-editing strategy for the creation of single nucleotide changes at specific loci. Additionally, we report a novel counterselection strategy which allows creation of precisely edited fungal strains that contain no foreign DNA and are completely isogenic to the wild type. Together, these developments represent a scalable improvement in the precision and speed of genetic manipulation in M. oryzae and are likely to be broadly applicable to other fungal species

The associations between structural treatment characteristics and post-treatment functioning in compulsory residential youth care

Contains fulltext : 102944.pdf (publisher's version ) (Open Access)In 2005 a new compulsory residential treatment program was developed for adolescents in need for protection against themselves or their environment. The aim of the present study was to examine the association of structural treatment characteristics of this new residential treatment program (i.e., duration of treatment, discharge status, and group composition in terms of sex) with post-treatment functioning. Additionally, the number of pre-treatment risk factors was included in the model. A total of 301 adolescents (174 boys, 127 girls), with a mean age at time of admittance of 15.50 (SD = 1.26) participated in this study. The number of risk factors was derived from treatment files of the adolescents at time of entrance. Six months after discharge, adolescents participated in a telephone interview to measure ten post-treatment variables indicating how well they were doing. The results showed that duration of treatment was related to post-treatment living situation, in that adolescents who were in treatment for shorter durations were more likely to live on their own after treatment. For discharge status, findings suggested that adolescents who were regularly discharged had more frequent contact with their family; however, they also showed higher alcohol consumption 6 months after treatment. Group composition was related to the girls' official offending, indicating that girls placed in mixed-sex groups showed significantly fewer official police contacts than did girls in girls-only treatment groups. Overall, structural treatment characteristics were hardly related to the adolescents' functioning after treatment. Suggestions for future research are discussed.20 p

The reliability and validity of the Dominic Interactive: A computerized child report instrument for mental health problems

Item does not contain fulltextChildren and youths' self-report of mental health problems is considered essential but complicated. This study examines the psychometric properties of the Dominic Interactive, a computerized DSM-IV based self-report questionnaire and explores informant correspondence. The Dominic Interactive was administered to 214 Dutch children, 6-11 year old, 122 attended special education schools and 92 children attended public schools. Within 2 weeks 155 children were reassessed. Parents of 211 children completed a paper version of the Dominic Interactive. The findings showed moderate to good internal consistency and test-retest reliability, although stability increased with age. Factor structure concerning generalized anxiety disorder and major depressive disorder displayed ambiguous results. Comparing special education with public school children revealed that both parents and children in special education reported significantly more problems on almost all scales. The results on informant correspondence revealed that discrepancies in reports were not consistent across individual pairs; they were due to a few extreme cases. Eliminating these cases increased the agreement in reports. Although more research is needed to determine the value of the Dominic Interactive, the Dominic Interactive has the potential to contribute with its unique features to the existing self-report instruments for screening and assessing child mental health.18 p

The braincase of a specimen of Proterochampsa Reig (Archosauriformes: Proterochampsidae) from the Late Triassic of Argentina

The proterochampsids are a Triassic group of superficially crocodile-like forms belonging to the Archosauriformes. In the present contribution, we present new information regarding the braincase of the proterochampsid Proterochampsa Reig 1959, from the Ischigualasto Formation (Carnian) of Argentina, and discuss its phylogenetic considerations. Some unique neurocranial features of Proterochampsa are described, including: the prominence and thickness of the V-shaped ridge that surrounds the basisphenoidal fossa; the medially concave lateral arms of the same ridge; and the semilunar depression on the parabasisphenoid ventrolaterally exposed. Other features are only shared with likely unrelated archosauriforms, including: the great lateral development of the basipterygoid processes and caudal development of its distal end; an eight-shaped metotic foramen; laterally directed basipterygoid processes; and rostral boundary of the basisphenoidal recess V-shaped. Proterochampsa differs in many other aspects from the archosauriform Chanaresuchus, including: a proportionally shorter basioccipital basal tubera; cultriform process ovoid in cross-section; longitudinal sulcus dorsal to the basipterygoid process; deep basisphenoidal recess; and the absence of a prominent intertuberal plate. In many braincase features, Proterochampsa is more similar to archosaurs than to Euparkeria, erythrosuchids and Proterosuchus. They include a reduced semilunar depression. A ventral border of the basioccipital forming a wide convexity and a dorsoventrally thin paroccipital process likely represents a feature shared with Chanaresuchus, but not with Doswellia and other basal archosauriforms.Fil: Trotteyn, Maria Jimena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Juan; Argentina. Universidad Nacional de San Juan; ArgentinaFil: Haro, Jose Augusto. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de San Juan; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Centro de Investigaciones Paleobiológicas; Argentin