Talking and Making Meaning About Parental Mental Health Problems: the Role of Children's Family Caregivers

When parents with severe and enduring mental health problems (MHP) are less able to meet their children’s needs, other family members often help with childcare. Research with children and parents with MHP has pointed to the vital role family caregivers play in helping children make meaning and communicate about their parents’ MHP. The caregiver’s perspective on this, however, is barely known. The purpose of this study was to examine how children’s family caregivers contribute to children’s meaning-making about parental MHP. In-depth interviews were conducted with 19 adults living in England and Wales who supported at least one related child aged 4-17 who had a parent with MHP. A substantive theory was developed using constructivist Grounded Theory. It was found that participants engaged in a core social process of providing protection in uncertainty, through which they sought to protect the child, the parent, the self, and the wider family. In making meaning and communicating with children, participants were concerned with shaping the interactional space, communicating through the developmental process, and engendering a sense of safety. Participants appeared to position themselves to the childcare role by assessing the child’s needs, the parent’s needs and their own needs. This was conceptualised as developing a caregiver identity, and seen to shape their contributions to children’s meaning-making. A large amount of communication was found to occur nonverbally, and seemingly without caregiver intent or awareness. The findings support the agenda for family-focused provision of mental health and social care. Clinical recommendations are made for better psychoeducation for caregivers and their inclusion in interventions with children and parents with MHP

Critical analysis of the governance of the Sainte Luce Locally Managed Marine Area (LMMA), southeast Madagascar

The Marine Protected Area Governance (MPAG) framework is applied to critically assess the governance of the Sainte Luce Locally Managed Marine Area (LMMA), southeast Madagascar. Madagascar experiences rapid population growth, widespread poverty, corruption and political instability, which hinders natural resource governance. Community-based natural resource management (CBNRM) has been repeatedly employed to circumvent the lack of state capacity. This includes the LMMA model, which has rapidly proliferated, represented by MIHARI, Madagascar's LMMA network. The lobster fishing is the primary source of income for households in the impoverished community of Sainte Luce, one of the key landing sites in the regional export industry. However, fishers, industry actors and available data suggest a significant decline of local and regional stocks, likely due to over-exploitation driven by poverty and migration. In 2013, SEED Madagascar a UK NGO, worked to establish community-based fishery management in Sainte Luce, setting up a local management committee, which introduced a periodic no take zone (NTZ). Despite the community's efforts and some significant achievements, the efficacy of management is limited. To date, limited state support and the lack of engagement by actors throughout the value chain have hampered effective governance. The study reinforces the finding that resilient governance relies on a diversity of actors and the incentives they collectively employ. Here and elsewhere, there is a limit to what can be achieved by bottom-up approaches in isolation. Resilient management of marine resources in Madagascar relies on improving the capacity of community, state, NGO and industry actors to collectively govern resources

Mitochondrially targeted ZFNs for selective degradation of pathogenic mitochondrial genomes bearing large‐scale deletions or point mutations

We designed and engineered mitochondrially targeted obligate heterodimeric zinc finger nucleases (mtZFNs) for site‐specific elimination of pathogenic human mitochondrial DNA (mtDNA). We used mtZFNs to target and cleave mtDNA harbouring the m.8993T>G point mutation associated with neuropathy, ataxia, retinitis pigmentosa (NARP) and the “common deletion” (CD), a 4977‐bp repeat‐flanked deletion associated with adult‐onset chronic progressive external ophthalmoplegia and, less frequently, Kearns‐Sayre and Pearson's marrow pancreas syndromes. Expression of mtZFNs led to a reduction in mutant mtDNA haplotype load, and subsequent repopulation of wild‐type mtDNA restored mitochondrial respiratory function in a CD cybrid cell model. This study constitutes proof‐of‐principle that, through heteroplasmy manipulation, delivery of site‐specific nuclease activity to mitochondria can alleviate a severe biochemical phenotype in primary mitochondrial disease arising from deleted mtDNA species

Genome editing in mitochondria corrects a pathogenic mtDNA mutation in vivo.

Mutations of the mitochondrial genome (mtDNA) underlie a substantial portion of mitochondrial disease burden. These disorders are currently incurable and effectively untreatable, with heterogeneous penetrance, presentation and prognosis. To address the lack of effective treatment for these disorders, we exploited a recently developed mouse model that recapitulates common molecular features of heteroplasmic mtDNA disease in cardiac tissue: the m.5024C>T tRNAAla mouse. Through application of a programmable nuclease therapy approach, using systemically administered, mitochondrially targeted zinc-finger nucleases (mtZFN) delivered by adeno-associated virus, we induced specific elimination of mutant mtDNA across the heart, coupled to a reversion of molecular and biochemical phenotypes. These findings constitute proof of principle that mtDNA heteroplasmy correction using programmable nucleases could provide a therapeutic route for heteroplasmic mitochondrial diseases of diverse genetic origin

The Atrial Fibrillation Risk Score for Hyperthyroidism Patients

Thyrotoxicosis (TT) is associated with an increase in both total and cardiovascu-lar mortality. One of the main thyrotoxicosis risks is Atrial Fibrillation (AF). Right AF predicts help medical personal prescribe the correct medicaments and correct surgical or radioiodine therapy. The main goal of this study is creating a method for practical treatment and diagnostic AF. This study proposes a new method for assessing the risk of occurrence atrial fibrillation for patients with TT. This method considers both the features of the complication and the specifics of the chronic disease. A model is created based on case histories of patients with thyrotoxicosis. We used Machine Learning methods for creating several models. Each model has advantages and disadvantages depending on the diagnostic and medical purposes. The resulting models show high results in the different metrics of the prediction of AF. These models interpreted and simple for use. Therefore, models can be used as part of the support and decision-making system (DSS) by medical specialists in the treatment and diagnostic of AF

Heterozygous SSBP1 start loss mutation co-segregates with hearing loss and the m.1555A>G mtDNA variant in a large multigenerational family.

The m.1555A>G mtDNA variant causes maternally inherited deafness, but the reasons for the highly variable clinical penetrance are not known. Exome sequencing identified a heterozygous start loss mutation in SSBP1, encoding the single stranded binding protein 1 (SSBP1), segregating with hearing loss in a multi-generational family transmitting m.1555A>G, associated with mtDNA depletion and multiple deletions in skeletal muscle. The SSBP1 mutation reduced steady state SSBP1 levels leading to a perturbation of mtDNA metabolism, likely compounding the intra-mitochondrial translation defect due to m.1555A>G in a tissue-specific manner. This family demonstrates the importance of rare trans-acting genetic nuclear modifiers in the clinical expression of mtDNA disease

Studies Directed Toward Improving the Spatial Resolution of the Distribution of Plutonium in Bone

Of several methods which have been discussed for the improvement of resolution in fission fragment track detectors for neutron-induced autoradiography, emphasis had been on the use of absorber layers inserted between the sample and the detector. Scanning electron microscopy was shown to be beneficial for viewing only the entrance holes of fission fragment tracks in the detector foil. The primary disadvantage of using thick absorbers lies in the factor of 50 to 100 percent loss in sensitivity over bare detectors. One promising solution to this problem is the use of glass detectors with high critical angles, no absorbers, and short etching times. Such detectors gave the best resolution of any system tested (+-2 $mu$), though they suffered at fluences greater than 10$sup 16$ n/sub th/ cm$sup 2$ from high backgrounds as a result of their natural uranium content. Two samples of bone, each of 1 mg were dissolved in HNO$sub 3$ spiked with $sup 244$Pu, and analyzed for total $sup 239$Pu content in an isotopic abundance mass spectrometer. The $sup 239$Pu concentration was 1 ppm by weight; bones with 10 ppm of $sup 239$Pu are required, therefore, for neutron-induced autoradiography. Bones in suitable form for IMMA required only to be coated with a thin conductive coating of gold. In a fast scan mode, all elements were searched in 200 x 200 areas. Good quality micrographs were obtained showing Na, K, Ca, and P as major constituents with minor elements present, such as Cr and Fe. (CH

Near-complete elimination of mutant mtDNA by iterative or dynamic dose-controlled treatment with mtZFNs.

Mitochondrial diseases are frequently associated with mutations in mitochondrial DNA (mtDNA). In most cases, mutant and wild-type mtDNAs coexist, resulting in heteroplasmy. The selective elimination of mutant mtDNA, and consequent enrichment of wild-type mtDNA, can rescue pathological phenotypes in heteroplasmic cells. Use of the mitochondrially targeted zinc finger-nuclease (mtZFN) results in degradation of mutant mtDNA through site-specific DNA cleavage. Here, we describe a substantial enhancement of our previous mtZFN-based approaches to targeting mtDNA, allowing near-complete directional shifts of mtDNA heteroplasmy, either by iterative treatment or through finely controlled expression of mtZFN, which limits off-target catalysis and undesired mtDNA copy number depletion. To demonstrate the utility of this improved approach, we generated an isogenic distribution of heteroplasmic cells with variable mtDNA mutant level from the same parental source without clonal selection. Analysis of these populations demonstrated an altered metabolic signature in cells harbouring decreased levels of mutant m.8993T>G mtDNA, associated with neuropathy, ataxia, and retinitis pigmentosa (NARP). We conclude that mtZFN-based approaches offer means for mtDNA heteroplasmy manipulation in basic research, and may provide a strategy for therapeutic intervention in selected mitochondrial diseases.Medical Research Council, UK; EMBO Fellowship [ALTF 701-2013 to L.V.H.]; PhD fellowship from the Foundation for Science and Technology, Portugal through the GABBA Program, University of Porto (to P.R.G.); Experiments undertaken in the J-PC laboratory were supported by ANR Investissement d’Avenir [ANR-IIINSB-0014] and AFM [18566].This is the final version of the article. It first appeared from Oxford University Press via http://dx.doi.org/10.1093/nar/gkw67

Heterozygous SSBP1 start loss mutation co-segregates with hearing loss and the m.1555A>G mtDNA variant in a large multigenerational family

The m.1555A>G mtDNA variant causes maternally inherited deafness, but the reasons for the highly variable clinical penetrance are not known. Exome sequencing identified a heterozygous start loss mutation in SSBP1, encoding the single stranded binding protein 1 (SSBP1), segregating with hearing loss in a multi-generational family transmitting m.1555A>G, associated with mtDNA depletion and multiple deletions in skeletal muscle. The SSBP1 mutation reduced steady state SSBP1 levels leading to a perturbation of mtDNA metabolism, likely compounding the intra-mitochondrial translation defect due to m.1555A>G in a tissue-specific manner. This family demonstrates the importance of rare trans-acting genetic nuclear modifiers in the clinical expression of mtDNA disease

Near-complete elimination of mutant mtDNA by iterative or dynamic dose-controlled treatment with mtZFNs

Mitochondrial diseases are frequently associated with mutations in mitochondrial DNA (mtDNA). In most cases, mutant and wild-type mtDNAs coexist, resulting in heteroplasmy. The selective elimination of mutant mtDNA, and consequent enrichment of wild-type mtDNA, can rescue pathological phenotypes in heteroplasmic cells. Use of the mitochondrially targeted zinc finger-nuclease (mtZFN) results in degradation of mutant mtDNA through site-specific DNA cleavage. Here, we describe a substantial enhancement of our previous mtZFN-based approaches to targeting mtDNA, allowing near-complete directional shifts of mtDNA heteroplasmy, either by iterative treatment or through finely controlled expression of mtZFN, which limits off-target catalysis and undesired mtDNA copy number depletion. To demonstrate the utility of this improved approach, we generated an isogenic distribution of heteroplasmic cells with variable mtDNA mutant level from the same parental source without clonal selection. Analysis of these populations demonstrated an altered metabolic signature in cells harbouring decreased levels of mutant m.8993T>G mtDNA, associated with neuropathy, ataxia, and retinitis pigmentosa (NARP). We conclude that mtZFN-based approaches offer means for mtDNA heteroplasmy manipulation in basic research, and may provide a strategy for therapeutic intervention in selected mitochondrial diseases