A putative silencer variant in a spontaneous canine model of retinitis pigmentosa

Author summary Retinitis pigmentosa (RP) is a blinding eye disease that affects nearly two million people worldwide. Several genes and variants have been associated with the disease, but still 30-80% of the patients lack genetic diagnosis. There is currently no standard treatment for RP, and much is expected from gene therapy. A similar disease, called progressive retinal atrophy (PRA), affects many dog breeds. We performed clinical, genetic and functional analyses to find the genetic cause for PRA in Miniature Schnauzers. We discovered two forms of PRA in the breed, named type 1 and 2, and show that they are genetically distinct as they map to different chromosomes, 15 and X, respectively. Further genetic, bioinformatic and functional analyses discovered a fully penetrant recessive variant in a putative silencer region for type 1 PRA. Silencer regions are important for gene regulation and we found that two of its predicted target genes, EDN2 and COL9A2, were overexpressed in the retina of the affected dog. Defects in both EDN2 and COL9A2 have been associated with retinal degeneration. This study provides new insights to retinal biology while the genetic test guides better breeding choices. Retinitis pigmentosa (RP) is the leading cause of blindness with nearly two million people affected worldwide. Many genes have been implicated in RP, yet in 30-80% of the RP patients the genetic cause remains unknown. A similar phenotype, progressive retinal atrophy (PRA), affects many dog breeds including the Miniature Schnauzer. We performed clinical, genetic and functional experiments to identify the genetic cause of PRA in the breed. The age of onset and pattern of disease progression suggested that at least two forms of PRA, types 1 and 2 respectively, affect the breed, which was confirmed by genome-wide association study that implicated two distinct genomic loci in chromosomes 15 and X, respectively. Whole-genome sequencing revealed a fully segregating recessive regulatory variant in type 1 PRA. The associated variant has a very recent origin based on haplotype analysis and lies within a regulatory site with the predicted binding site of HAND1::TCF3 transcription factor complex. Luciferase assays suggested that mutated regulatory sequence increases expression. Case-control retinal expression comparison of six best HAND1::TCF3 target genes were analyzed with quantitative reverse-transcriptase PCR assay and indicated overexpression of EDN2 and COL9A2 in the affected retina. Defects in both EDN2 and COL9A2 have been previously associated with retinal degeneration. In summary, our study describes two genetically different forms of PRA and identifies a fully penetrant variant in type 1 form with a possible regulatory effect. This would be among the first reports of a regulatory variant in retinal degeneration in any species, and establishes a new spontaneous dog model to improve our understanding of retinal biology and gene regulation while the affected breed will benefit from a reliable genetic testing.Peer reviewe

E.U. paediatric MOG consortium consensus: Part 4 - Outcome of paediatric myelin oligodendrocyte glycoprotein antibody-associated disorders

There is increasing knowledge on the role of antibodies against myelin oligodendrocyte glycoprotein (MOG-abs) in acquired demyelinating syndromes and autoimmune encephalitis in children. Better understanding and prediction of outcome is essential to guide treatment protocol decisions. Therefore, this part of the Paediatric European Collaborative Consensus provides an oversight of existing knowledge of clinical outcome assessment in paediatric MOG-ab-associated disorders (MOGAD). The large heterogeneity in disease phenotype, disease course, treatment and follow-up protocols is a major obstacle for reliable prediction of outcome. However, the clinical phenotype of MOGAD appears to be the main determinant of outcome. Patients with a transverse myelitis phenotype in particular are at high risk of accruing neurological disability (motor and autonomic), which is frequently severe. In contrast, having a single episode of optic neuritis any time during disease course is broadly associated with a lower risk of persistent disability. Furthermore, MOG-ab-associated optic neuritis often results in good functional visual recovery, although retinal axonal loss may be severe. The field of cognitive and behavioural outcome and epilepsy following demyelinating episodes has not been extensively explored, but in recent studies acute disseminated encephalomyelitis (-like) phenotype in the young children was associated with cognitive problems and epilepsy in long-term follow-up. In conclusion, main domains of importance in determining clinical outcome in paediatric MOGAD are visual, motor, autonomic and cognitive function. A standardised evaluation of these outcome domains in all children is of importance to allow adequate rehabilitation and follow-up

The SNARE Protein SNAP23 and the SNARE-Interacting Protein Munc18c in Human Skeletal Muscle Are Implicated in Insulin Resistance/Type 2 Diabetes

OBJECTIVE-Our previous studies suggest that the SNARE protein synaptosomal-associated protein of 23 kDa (SNAP23) is involved in the link between increased lipid levels and insulin resistance in cardiomyocytes. The objective was to determine whether SNAP23 may also be involved in the known association between lipid accumulation in skeletal muscle and insulin resistance/type 2 diabetes in humans, as well as to identify a potential regulator of SNAP23. RESEARCH DESIGN AND METHODS-We analyzed skeletal muscle biopsies from patients with type 2 diabetes and healthy, insulin-sensitive control subjects for expression (mRNA and protein) and intracellular localization (subcellular fractionation and immunohistochemistry) of SNAP23, and for expression of proteins known to interact with SNARE proteins. Insulin resistance was determined by a euglycemic hyperinsulinemic clamp Potential mechanisms for regulation of SNAP23 were also investigated in the skeletal muscle cell line L6. RESULTS-We showed increased SNAP23 levels in skeletal muscle from patients with type 2 diabetes compared with that from lean control subjects Moreover, SNAP23 was redistributed from the plasma membrane to the microsomal/cytosolic compartment in the patients with the type 2 diabetes Expression of the SNARE-interacting protein Munc18c was higher in skeletal muscle from patients with type 2 diabetes Studies in L6 cells showed that Munc18c promoted the expression of SNAP23. CONCLUSIONS-We have translated our previous in vitro results into humans by showing that there is a change in the distribution of SNAP23 to the interior of the cell in skeletal muscle from patients with type 2 diabetes. We also showed that Munc18c is a potential regulator of SNAP23. Diabetes 59: 1870-1878, 201

Current international trends in the treatment of multiple sclerosis in children:impact of the COVID-19 pandemic

Background: Only recently has the first disease-modifying therapy been approved for children with multiple sclerosis (MS) and practice patterns including substantial off-label use have evolved. Understanding attitudes towards treatment of paediatric MS and whether this has changed due to the ongoing COVID-19 pandemic is vital to guide future therapeutic trials and for developing guidelines that reflect practice. Methods: We performed an online survey within the International Paediatric Multiple Sclerosis Study Group between July and September 2020. The survey was sent to 130 members from 25 countries and consisted of five sections: demographic data, treatment, disease modifying therapies and COVID-19, outcome and three patient cases. Results: The survey was completed by 66 members (51%), both paediatric neurologists and adult neurologists. Fingolimod and β-interferons were the most frequently used disease-modifying therapies, especially among paediatric neurologists. Almost a third (31%) of respondents had altered their prescribing practice due to COVID-19, in particular at the beginning of the pandemic. Conclusions: The survey results indicate a tendency of moving from the traditional escalation therapy starting with injectables towards an early start with newer, highly effective disease modifying therapies. The COVID-19 pandemic only slightly affected prescribing patterns and treatment choices in paediatric MS

The multiple functions of miR-574-5p in the neuroblastoma tumor microenvironment

Neuroblastoma is the most common extracranial solid tumor in childhood and arises from neural crest cells of the developing sympathetic nervous system. Prostaglandin E2 (PGE2) has been identified as a key pro-inflammatory mediator of the tumor microenvironment (TME) that promotes neuroblastoma progression. We report that the interaction between the microRNA miR-574-5p and CUG-binding protein 1 (CUGBP1) induces the expression of microsomal prostaglandin E2 synthase 1 (mPGES-1) in neuroblastoma cells, which contributes to PGE2 biosynthesis. PGE2 in turn specifically induces the sorting of miR-574-5p into small extracellular vesicles (sEV) in neuroblastoma cell lines. sEV are one of the major players in intercellular communication in the TME. We found that sEV-derived miR-574-5p has a paracrine function in neuroblastoma. It acts as a direct Toll-like receptor 7/8 (TLR7/8) ligand and induces α-smooth muscle actin (α-SMA) expression in fibroblasts, contributing to fibroblast differentiation. This is particularly noteworthy as it has an opposite function to that in the TME of lung carcinoma, another PGE2 dependent tumor type. Here, sEV-derived miR-574-5p has an autokrine function that inhibits PGE2 biosynthesis in lung cancer cells. We report that the tetraspanin composition on the surface of sEV is associated with the function of sEV-derived miR-574-5p. This suggests that the vesicles do not only transport miRs, but also appear to influence their mode of action

Identification of a Novel Topoisomerase Inhibitor Effective in Cells Overexpressing Drug Efflux Transporters

BACKGROUND:Natural product structures have high chemical diversity and are attractive as lead structures for discovery of new drugs. One of the disease areas where natural products are most frequently used as therapeutics is oncology. METHOD AND FINDINGS:A library of natural products (NCI Natural Product set) was screened for compounds that induce apoptosis of HCT116 colon carcinoma cells using an assay that measures an endogenous caspase-cleavage product. One of the apoptosis-inducing compounds identified in the screen was thaspine (taspine), an alkaloid from the South American tree Croton lechleri. The cortex of this tree is used for medicinal purposes by tribes in the Amazonas basin. Thaspine was found to induce conformational activation of the pro-apoptotic proteins Bak and Bax, mitochondrial cytochrome c release and mitochondrial membrane permeabilization in HCT116 cells. Analysis of the gene expression signature of thaspine-treated cells suggested that thaspine is a topoisomerase inhibitor. Inhibition of both topoisomerase I and II was observed using in vitro assays, and thaspine was found to have a reduced cytotoxic effect on a cell line with a mutated topoisomerase II enzyme. Interestingly, in contrast to the topoisomerase II inhibitors doxorubicin, etoposide and mitoxantrone, thaspine was cytotoxic to cell lines overexpressing the PgP or MRP drug efflux transporters. We finally show that thaspine induces wide-spread apoptosis in colon carcinoma multicellular spheroids and that apoptosis is induced in two xenograft mouse models in vivo. CONCLUSIONS:The alkaloid thaspine from the cortex of Croton lechleri is a dual topoisomerase inhibitor effective in cells overexpressing drug efflux transporters and induces wide-spread apoptosis in multicellular spheroids

Relation of academic motivation and attitude towards studies of students of High Scholl X

Sweden has formulated ambitious energy goals which signify that by year 2050 the energy usage should be halved. One of the objectives is the reduction of energy usage by 20% by year 2020 and that it should be compared to the usage of energy from 1995. In parallel with increased energy requirements for buildings the demand on housing conservation, by highlighting also buildings from the last 50 years as part of the building heritage, continues to enhance, which in theory means that there is a conflict between these. The law will never be able to point out unique buildings and PBL (planning and building act) is not especially concrete. The National Housing Board has tried to make this more concrete by the new BBR regulations though there are still an enormous variety of interpretations. This report deals with the conflict between conservation requirements and additional insulation of the building envelope and is limited to multi-residential buildings built between the years 1945 and 1975. It is the stock with the greatest energy consumption and that is where the big challenge lies. The question is whether in practice it is no conflict between conservation requirements and additional insulation of the building envelope and what it in this case depends. With the support of background facts, interview questions were formulated for different participants. By interviews with property-owners, municipalities and other partners it has been shown that the creation of zoning determines whether a property has been preserved. In municipalities with a metropolitan character the interest in conservation issues are the greatest and Stockholm municipality distinguishes itself, providing significant space in regards to this subject. The smaller municipalities do not believe in that the preservation of our multi-residential buildings are central but that there lies a greater problem in areal density and that eventually great portions of the genuine country will disappear. Only in isolated cases, conservation requirements became an obstacle in performing façade renovations. Property owners are interested in the conservation issue; however, it must be weighed against the economic condition and therefore is it in sight of what is a good conservation.Sverige har formulerat ambitiösa energimål som innebär att till år 2050 ska energianvändningen halveras, där ett delmål är minskning med 20 % till år 2020 och det ska jämföras med energianvändningen från år 1995. Parallellt med ökade energikrav för byggnader ställs det även allt högre krav på husens bevarande, vilket teoretiskt innebär att det ligger en konflikt mellan dessa. Lagen kommer aldrig att kunna peka ut unika byggnader och PBL (plan- och bygglagen) är inte särskilt konkret. Nu har Boverket försökt konkretisera genom nya BBR reglerna, men det finns ändå oerhört mycket plats för tolkningar. Denna rapport behandlar konflikten mellan bevarandekrav och tilläggsisolering av klimatskalet och är begränsad till flerbostadshus byggda mellan åren 1945 och 1975. Det är det bestånd som har störst energiåtgång och det är där den stora utmaningen ligger. Frågan är om det i praktiken ligger någon konflikt mellan bevarandekrav och tilläggsisolering av klimatskalet och vad den i så fall beror på? Med stöd av bakgrundsfakta har intervjufrågor formulerats till olika aktörer. I intervjuer med fastighetsägare, kommuner och övriga parter har det visat sig att utformningen av detaljplanen är avgörande för om en fastighet kan bevaras. I kommuner med storstadskaraktär är intresset för bevarandefrågorna störst och Stockholms kommun utmärker sig, där dessa frågor getts störst utrymme. De mindre kommunerna anser inte att bevarandet av våra flerbostadshus är centralt utan där är det ett större problem att områdena förtätas och att på sikt stora delar av den genuina landsbygden kommer att försvinna. Endast i enstaka fall har bevarandekraven blivit ett hinder för att utföra fasadrenoveringar. Fastighetsägare är intresserade för bevarande frågan men detta måste vägas mot ekonomiska förutsättningar och därför skiljer det i synen om vad som är ett gott bevarand

Konflikten mellan bevarandekrav och tilläggsisolering : The conflict between conservation requirements and additional insulation

Sweden has formulated ambitious energy goals which signify that by year 2050 the energy usage should be halved. One of the objectives is the reduction of energy usage by 20% by year 2020 and that it should be compared to the usage of energy from 1995. In parallel with increased energy requirements for buildings the demand on housing conservation, by highlighting also buildings from the last 50 years as part of the building heritage, continues to enhance, which in theory means that there is a conflict between these. The law will never be able to point out unique buildings and PBL (planning and building act) is not especially concrete. The National Housing Board has tried to make this more concrete by the new BBR regulations though there are still an enormous variety of interpretations. This report deals with the conflict between conservation requirements and additional insulation of the building envelope and is limited to multi-residential buildings built between the years 1945 and 1975. It is the stock with the greatest energy consumption and that is where the big challenge lies. The question is whether in practice it is no conflict between conservation requirements and additional insulation of the building envelope and what it in this case depends. With the support of background facts, interview questions were formulated for different participants. By interviews with property-owners, municipalities and other partners it has been shown that the creation of zoning determines whether a property has been preserved. In municipalities with a metropolitan character the interest in conservation issues are the greatest and Stockholm municipality distinguishes itself, providing significant space in regards to this subject. The smaller municipalities do not believe in that the preservation of our multi-residential buildings are central but that there lies a greater problem in areal density and that eventually great portions of the genuine country will disappear. Only in isolated cases, conservation requirements became an obstacle in performing façade renovations. Property owners are interested in the conservation issue; however, it must be weighed against the economic condition and therefore is it in sight of what is a good conservation.Sverige har formulerat ambitiösa energimål som innebär att till år 2050 ska energianvändningen halveras, där ett delmål är minskning med 20 % till år 2020 och det ska jämföras med energianvändningen från år 1995. Parallellt med ökade energikrav för byggnader ställs det även allt högre krav på husens bevarande, vilket teoretiskt innebär att det ligger en konflikt mellan dessa. Lagen kommer aldrig att kunna peka ut unika byggnader och PBL (plan- och bygglagen) är inte särskilt konkret. Nu har Boverket försökt konkretisera genom nya BBR reglerna, men det finns ändå oerhört mycket plats för tolkningar. Denna rapport behandlar konflikten mellan bevarandekrav och tilläggsisolering av klimatskalet och är begränsad till flerbostadshus byggda mellan åren 1945 och 1975. Det är det bestånd som har störst energiåtgång och det är där den stora utmaningen ligger. Frågan är om det i praktiken ligger någon konflikt mellan bevarandekrav och tilläggsisolering av klimatskalet och vad den i så fall beror på? Med stöd av bakgrundsfakta har intervjufrågor formulerats till olika aktörer. I intervjuer med fastighetsägare, kommuner och övriga parter har det visat sig att utformningen av detaljplanen är avgörande för om en fastighet kan bevaras. I kommuner med storstadskaraktär är intresset för bevarandefrågorna störst och Stockholms kommun utmärker sig, där dessa frågor getts störst utrymme. De mindre kommunerna anser inte att bevarandet av våra flerbostadshus är centralt utan där är det ett större problem att områdena förtätas och att på sikt stora delar av den genuina landsbygden kommer att försvinna. Endast i enstaka fall har bevarandekraven blivit ett hinder för att utföra fasadrenoveringar. Fastighetsägare är intresserade för bevarande frågan men detta måste vägas mot ekonomiska förutsättningar och därför skiljer det i synen om vad som är ett gott bevarand

preoperative anxiety : Non-pharmacological strategies to help troubled children-A systematic literature study

Sammanfattning Bakgrund:  Preoperativ oro kan orsaka höga nivåer av stress hos barnet bland annat på grund av separation från föräldrarna, rädsla, inte veta vad som skall hända och att befinna sig i en okänd miljö. Upp till 50% av barn som skall opereras upplever preoperativ oro vilket kan leda till oönskade konsekvenser som högre förekomst av postoperativt delirium, postoperativ oro och ökad smärta men även ge negativa effekter under en längre tid postoperativt som mardrömmar, att vakna gråtandes, separationsångest, vredesutbrott, trots riktat mot föräldrarna och nytillkommen nattlig sängvätning. Anestesisjuksköterskan har ett ansvar i att minska preoperativ oro och därmed förbättra barnets upplevelse av situationen och förebygga negativa konsekvenser. Syfte: Syftet med studien är att belysa icke farmakologiska strategier och deras effekt i arbetet med preoperativt oroliga barn vilka skall genomgå generell anestesi. Metod: Systematisk litteraturstudie. Inklusionskriterier experiment studier rörande barn 0-18 år som skulle genomgå generell anestesi inför operation,  publicerade 120101-180213, i vetenskapliga engelskspråkiga referee-granskade tidskrifter. Databassökningar utfördes i PubMED, Cinahl och PsychINFO. Resultat: Två metoder som effektivt lindrar oro framkom i materialet. Distraktion, genom film, spel eller böcker, i samband med anestesiinduktion är en metod. Även preoperativt förberedande information i form av multi-media eller specialskrivna texter (sagoböcker och informationsbroschyrer) har visat sig ha en lindrande effekt. Utvidgad information till föräldrar, med tanke att förberedda föräldrar bättre hjälper sitt barn och minskar barnets preoperativa oro, gav inte mätbar effekt på barnens oro.Abstract Background: Preoperative anxiety can cause high levels of stress in the child, for example due to separation from parents, fear, not knowing what to happen and being in an unknown environment. Up to 50% of children to be treated experience preoperative anxiety which may lead to unwanted consequences such as higher incidence of postoperative delirium, postoperative anxiety and increased pain but also give negative effects for a longer period of time postoperatively like nightmares, waking crying, separation anxiety, outbreaks of anger, temper tantrums and enuresis. Nurse anesthetist has a responsibility in reducing preoperative anxiety and thus improving the child's experience of the situation, preventing negative consequences. Purpose: The purpose of the study is to highlight non-pharmacological strategies and their effects in the work with preoperatively anxoius children, undergoing general anesthesia. Method: Systematic literature study. Inclusion criteria experimental studies on children 0-18 years who were to undergo general anesthesia prior to surgery, published 120101-180213, in scientific English-language referee-reviewed journals. Database searches were performed in PubMED, Cinahl and PsychINFO. Result: Two methods that relieve anxiety emerged in the material. Distraction, through movies, games or books, during the anesthesia indiction or preoperatively preparatory information in the form of multi-media or special-written texts (storybooks and information brochures). Expanded information to parents, given that prepared parents better help their children and reduce the child's preoperative anxiety, did not give a measurable effect. Conclusion: Preoperative, age-appropriate information for the child and distraction during the anesthesia induction are, respectively, effective methods to alleviate the preoperative anxiety of children

How can sexual and reproductive health and rights be enhanced for young people with intellectual disability? : focus group interviews with staff in Sweden

Background Different types of staff support individuals with intellectual disability (ID) in their daily life, in schools, leisure activities and in special accommodations. This study aimed to gain a deeper understanding of experiences and perceptions regarding sexual and reproductive health and rights (SRHR) among staff. Methods Data were collected in mid-Sweden in four focus groups with altogether 20 participants, 18 women and 2 men aged between 18 and 65 years. They had different professions and worked among youth and adults with ID aged 18–40 years in schools, accommodations and with leisure activities. Their working experience varied from 3 years to more than 20 years. Interviews were audio recorded, transcribed and analysed with content analysis. Results The participants generally described positive attitudes towards sexuality for people with ID, both among themselves and in society. However, many situations such as ensuring privacy, balancing between waiting and acting, issues around contraception and reproduction were difficult to address and participants had hesitations about childbearing. They described different strategies such as showing respect, enhancing self-esteem and decision making ability and using interprofessional support to cope with frustrating situations. They lacked a clear mandate from managers as well as written guidelines and policies. They requested education and support from peers, supervisors and other professionals. Conclusion Participants in the study were generally open-minded and accepting towards sexuality among young people with ID. They thought it was difficult to deal with reproduction/parenthood and felt unprepared and frustrated in certain situations. The participants requested a clear mandate from managers, organizational guidelines, more education and inter-professional support. We believe these findings can inform the development of policy and support the implementation of SRHR related guidelines to support staff working with young people with ID