A balancing act: integrating the expertise of youth peer workers in child and adolescent mental health services.

The socio-relational focus of youth peer support workers (YPSWs) poses a challenge when YPSWs are embedded in medical oriented contexts common to child and adolescent mental health services (CAMHS); as it requires YPSWs to find a balance between being a peer on one hand, and adhering to professional boundaries and medical standards set out by CAMHS on the other. To create a suitable position for YPSWs in CAMHS, this study investigated the unique socio-relational contributions YPSWs can make to CAMHS in addition to clinicians, and identified how these contributions can be embedded within CAMHS. This study reports on 37 semi-structured interviews conducted in the Netherlands with youth (n = 10), YPSWs (n = 10), and clinicians (n = 17). Overall, the unique socio-relational contributions YPSWs can make include: their ability to build authentic trusting relationships with youth by providing empowerment, promoting autonomy, valuing stillness in recovery, reducing isolation, recognizing strengths, and navigating life inside and outside of (residential) mental healthcare and beyond classification. Moreover, prerequisites to safeguard the integration of YPSWs and these socio-relational contributions were also identified, including YPSWs achieving stability in recovery, recent lived experiences with mental health challenges, and organizational support in terms of suitable treatment climate, resources to enhance flexibility of YPSWs, and shared goals regarding youth peer support work. Overall, YPSWs view youth holistically and foster a connection with youth based on youthfulness and recent lived experience. Involving YPSWs is an important step forward to drive positive transformation in CAMHS.

Multi-functional T-DNA/Ds tomato lines designed for gene cloning and molecular and physical dissection of the tomato genome.

In order to make the tomato genome more accessible for molecular analysis and gene cloning, we have produced 405 individual tomato (Lycopersicon esculentum) lines containing a characterized copy of pJasm13, a multifunctional T-DNA/modified Ds transposon element construct. Both the T-DNA and the Ds element in pJasm13 harbor a set of selectable marker genes to monitor excision and reintegration of Ds and additionally, target sequences for rare cutting restriction enzymes (I-PpoI, SfiI, NotI) and for site-specific recombinases (Cre, FLP, R). Blast analysis of flanking genomic sequences of 174 T-DNA inserts revealed homology to transcribed genes in 69 (40%), of which about half are known or putatively identified as genes and ESTs. The map position of 140 individual inserts was determined on the molecular genetic map of tomato. These inserts are distributed over the 12 chromosomes of tomato, allowing targeted and non-targeted transposon tagging, marking of closely linked genes of interest and induction of chromosomal rearrangements including translocations or creation of saturation-deletions or inversions within defined regions linked to the T-DNA insertion site. The different features of pJasm13 were successfully tested in tomato and Arabidopsis thaliana, thus providing a new tool for molecular/genetic dissection studies, including molecular and physical mapping, mutation analysis and cloning strategies in tomato and potentially, in other plants as well.

The tomato Mi-1 gene confers resistance to both root-knot nematodes and potato aphids.

Mi-1, a Lycopersicon peruvianum gene conferring resistance to the agricultural pests, root-knot nematodes, and introgressed into tomato, has been cloned using a selective restriction fragment amplification based strategy. Complementation analysis of a susceptible tomato line with a 100 kb cosmid array yielded a single cosmid clone capable of conferring resistance both to the root-knot nematode Meloidogyne incognita and to an unrelated pathogen, the potato aphid Macrosiphum euphorbiae. This resistance was stable. The Mi-1 gene encodes a protein sharing structural features with the nucleotide-binding site leucine-rich repeat-containing type of plant resistance genes.

Dissection of the fusarium I2 gene cluster in tomato reveals six homologs and one active gene copy.

The I2 locus in tomato confers resistance to race 2 of the soil-borne fungus Fusarium oxysporum f sp lycopersici. The selective restriction fragment amplification (AFLP) positional cloning strategy was used to identify I2 in the tomato genome. A yeast artificial chromosome (YAC) clone covering approximately 750 kb encompassing the I2 locus was isolated, and the AFLP technique was used to derive tightly linked AFLP markers from this YAC clone. Genetic complementation analysis in transgenic R1 plants using a set of overlapping cosmids covering the I2 locus revealed three cosmids giving full resistance to F. o. lycopersici race 2. These cosmids shared a 7-kb DNA fragment containing an open reading frame encoding a protein with similarity to the nucleotide binding site leucine-rich repeat family of resistance genes. At the I2 locus, we identified six additional homologs that included the recently identified I2C-1 and I2C-2 genes. However, cosmids containing the I2C-1 or I2C-2 gene could not confer resistance to plants, indicating that these members are not the functional resistance genes. Alignments between the various members of the I2 gene family revealed two significant variable regions within the leucine-rich repeat region. They consisted of deletions or duplications of one or more leucine-rich repeats. We propose that one or both of these leucine-rich repeats are involved in Fusarium wilt resistance with I2 specificity.

AFLP-based fine mapping of the Mlo gene to a 30-kb DNA segment of the barley genome.

Resistance of barley (Hordeum vulgare) to the powdery mildew fungus Erysiphe graminis f.sp. hordei is conferred by several dominant genes, but also by recessive alleles of the Mlo locus mapping on the long arm of chromosome 4. In addition, this single-factor-mediated resistance is active against all known physiological races of the parasite. Thus the mechanism underlying mlo-mediated resistance should differ substantially from that mediated by the dominant genes. A positional cloning strategy to isolate the Mlo gene from the barley genome, the size of which is almost double the size of the human genome, has been designed. The AFLP technique was employed to identify markers tightly linked to the Mlo locus and to produce a local high-resolution genetic map. The use of this high-volume marker technology allowed the rapid screening of approximately 250,000 loci for linkage to Mlo. A large number of Mlo-linked AFLP markers were identified, one of which cosegregated with Mlo on the basis of more than 4000 meiotic events. A four-genome-equivalent barley YAC library (average insert size 480 kb) was constructed and screened with this cosegregating marker. Four YACs containing this marker were isolated and subsequent characterization by AFLP-based physical mapping allowed the physical delimitation of the Mlo locus to a DNA segment of 30 kb.

The barley Mlo gene: a novel control element of plant pathogen resistance.

Mutation-induced recessive alleles (mlo) of the barley Mlo locus confer a leaf lesion phenotype and broad spectrum resistance to the fungal pathogen, Erysiphe graminis f. sp. hordei. The gene has been isolated using a positional cloning approach. Analysis of 11 mutagen-induced mlo alleles revealed mutations leading in each case to alterations of the deduced Mlo wild-type amino acid sequence. Susceptible intragenic recombinants, isolated from mlo heteroallelic crosses, show restored Mlo wild-type sequences. The deduced 60 kDa protein is predicted to be membrane-anchored by at least six membrane-spanning helices. The findings are compatible with a dual negative control function of the Mlo protein in leaf cell death and in the onset of pathogen defense; absence of Mlo primes the responsiveness for the onset of multiple defense functions.

Splicing features in maize streak virus virion- and complementary-sense gene expression.

The single-stranded DNA geminiviruses produce transcripts from both strands (virion- and complementary-sense) of a nuclear double-stranded DNA molecule. In maize streak virus (MSV)-infected maize plants, approximately 80% of the complementary-sense transcripts produce the C1 protein, whilst the remaining 20% are spliced to remove a 92 nt intron and produce a C1:C2 fusion protein (Rep). Disruption of the complementary-sense 3' splice site abolished virus replication. The majority of the virion-sense transcripts initiated one nucleotide upstream of the V1 (movement protein) gene and a minority a further 141 nucleotides upstream. A 76 nt intron, with features typical of plant introns, was identified within the V1 gene, upstream of the coat protein gene. Spliced and unspliced forms of each virion-sense transcript were produced, but they differed in splicing efficiency. Approximately 50% of the major transcript and less than 10% of the minor transcript were processed. Mutagenesis of the consensus 5' splice site in the V1 gene resulted in the use of alternative cryptic splice sites, confirming the importance of splicing for MSV infection. Spliced virion-sense transcripts were also identified in tissue infected with the closely-related Digitaria streak virus (DSV) but not with another subgroup I geminivirus, wheat dwarf virus. Collectively, the multiple transcript initiation sites and different splicing efficiencies suggest that splicing is an important feature in the regulation of both early and late gene expression in MSV and DSV.