Autism spectrum disorders (ASDs) are a group of devastating neurodevelopmental syndromes that affect up to 1 in 68 children. Despite advances in the identification of ASD risk genes, the mechanisms ...underlying ASDs remain unknown. Homozygous loss-of-function mutations in Contactin Associated Protein-like 2 (CNTNAP2) are strongly linked to ASDs. Here we investigate the function of Cntnap2 and undertake pharmacological screens to identify phenotypic suppressors. We find that zebrafish cntnap2 mutants display GABAergic deficits, particularly in the forebrain, and sensitivity to drug-induced seizures. High-throughput behavioral profiling identifies nighttime hyperactivity in cntnap2 mutants, while pharmacological testing reveals dysregulation of GABAergic and glutamatergic systems. Finally, we find that estrogen receptor agonists elicit a behavioral fingerprint anti-correlative to that of cntnap2 mutants and show that the phytoestrogen biochanin A specifically reverses the mutant behavioral phenotype. These results identify estrogenic compounds as phenotypic suppressors and illuminate novel pharmacological pathways with relevance to autism.
•Zebrafish mutants of the autism risk gene cntnap2 have GABAergic neuron deficits•High-throughput behavioral profiling identifies nighttime hyperactivity in mutants•cntnap2 mutants exhibit altered responses to GABAergic and glutamatergic compounds•Estrogenic compounds suppress the cntnap2 mutant behavioral phenotype
Hoffman et al. use zebrafish mutants of the autism risk gene Contactin Associated Protein-like 2 (CNTNAP2) to conduct high-throughput quantitative behavioral profiling and pharmacological screening. This approach reveals dysregulation of GABAergic and glutamatergic systems in mutants and identifies estrogenic compounds as suppressors of the mutant behavioral phenotype.
DNA methylation serves as an important epigenetic mark in both eukaryotic and prokaryotic organisms. In eukaryotes, the most common epigenetic mark is 5-methylcytosine, whereas prokaryotes can have ...6-methyladenine, 4-methylcytosine, or 5-methylcytosine. Single-molecule, real-time sequencing is capable of directly detecting all three types of modified bases. However, the kinetic signature of 5-methylcytosine is subtle, which presents a challenge for detection. We investigated whether conversion of 5-methylcytosine to 5-carboxylcytosine using the enzyme Tet1 would enhance the kinetic signature, thereby improving detection.
We characterized the kinetic signatures of various cytosine modifications, demonstrating that 5-carboxylcytosine has a larger impact on the local polymerase rate than 5-methylcytosine. Using Tet1-mediated conversion, we show improved detection of 5-methylcytosine using in vitro methylated templates and apply the method to the characterization of 5-methylcytosine sites in the genomes of Escherichia coli MG1655 and Bacillus halodurans C-125.
We have developed a method for the enhancement of directly detecting 5-methylcytosine during single-molecule, real-time sequencing. Using Tet1 to convert 5-methylcytosine to 5-carboxylcytosine improves the detection rate of this important epigenetic marker, thereby complementing the set of readily detectable microbial base modifications, and enhancing the ability to interrogate eukaryotic epigenetic markers.
Clinical ethics: medical tourism in children Birchley, Giles; Linney, Mike; Turner, Stephen W ...
Archives of disease in childhood,
12/2021, Volume:
106, Issue:
12
Journal Article
Peer reviewed
Open access
Decisions to seek treatment abroad are made privately by parents and usually uncontested by National Health Service (NHS) staff, despite some high-profile court cases. Treatment abroad often lacks ...continuity of care, with variable (or absent) sharing of medical information between countries if there are problems. Because medical treatment is expensive, crowdfunding websites have become a popular method of raising funds. The emphasis on public sympathy means that crowdfunding does not distribute resources equitably, and minority groups may raise less funds through crowdfunding than their non-minority peers.3 Crowdfunding publicises the private health information of a child with potentially lifelong impacts. Depending on what went wrong, it could be difficult to get problems treated on the NHS.5 Furthermore, even where treatment commenced overseas is successful it may not be funded on the child/young person’s return to the UK (although the best interests of the child will be the deciding factor in these cases).
BackgroundThe General and Adolescent Paediatric Research Network in the UK and Ireland (GAPRUKI) was established in 2016. The aims of GAPRUKI are to unite general paediatricians around the UK and ...Ireland, to develop research ideas and protocols, and facilitate delivery of multicentre research.ObjectivesTo undertake a research prioritisation exercise among UK and Ireland general paediatricians.MethodsThis was a four-phase study using a modified Delphi survey. The first phase asked for suggested research priorities. The second phase developed ideas and ranked them in priority. In the third phase, priorities were refined; and the final stage used the Hanlon Prioritisation Process to agree on the highest priorities.ResultsIn phase one, there were 250 questions submitted by 61 GAPRUKI members (66% of the whole membership). For phase two, 92 priorities were scored by 62 members and the mean Likert scale (1–7) scores ranged from 3.13 to 5.77. In a face-to-face meeting (phases three and four), 17 research questions were identified and ultimately 14 priorities were identified and ranked. The four priorities with the highest ranking focused on these three respiratory conditions: asthma, bronchiolitis and acute wheeze. Other priorities were in the diagnosis or management of constipation, urinary tract infection, fever, gastro-oesophageal reflux and also new models of care for scheduled general paediatric clinics.ConclusionResearch priorities for child health in the UK and Ireland have been identified using a robust methodology. The next steps are for studies to be designed and funded to address these priorities.
Quality of life: what matters? Hain, Richard D W; Turner, Stephen W
Archives of disease in childhood,
06/2022, Volume:
107, Issue:
6
Journal Article
Peer reviewed
National Institute for Health and Care Excellence (NICE) evaluates cost-effectiveness using a measure that combines the likelihood of prolonging an individual’s life and improving her QL: ‘QALYs ...(Quality-Adjusted Life Years) are calculated by estimating the years of life remaining for a patient and weighting each year with a quality-of-life score (on a 0 to 1 scale)’.8 NICE will usually recommend a treatment which costs £20 000–30 000 per QALY. The future benefits and harms of puberty-blocking drugs, for example, will extend over several decades and need to be appropriately weighted when considered alongside their effects on the child’s QL in the present moment. Since contributors to the quality of a child’s life, and its normal duration, are both different from those of an adult, it is perhaps surprising that there is no specifically paediatric QALY ‘tariff’ set by NICE. Summary Most paediatricians intuitively recognise that compassionate medical care should not restrict itself to curing a child’s condition but should also attend to enabling her to enjoy her life as much as possible (for example, treating enuresis to allow sleep-overs). The role of Response-Shift in studies assessing quality of life outcomes among cancer patients: a systematic review.
A novel template design for single-molecule sequencing is introduced, a structure we refer to as a SMRTbellTM template. This structure consists of a double-stranded portion, containing the insert of ...interest, and a single-stranded hairpin loop on either end, which provides a site for primer binding. Structurally, this format resembles a linear double-stranded molecule, and yet it is topologically circular. When placed into a single-molecule sequencing reaction, the SMRTbell template format enables a consensus sequence to be obtained from multiple passes on a single molecule. Furthermore, this consensus sequence is obtained from both the sense and antisense strands of the insert region. In this article, we present a universal method for constructing these templates, as well as an application of their use. We demonstrate the generation of high-quality consensus accuracy from single molecules, as well as the use of SMRTbell templates in the identification of rare sequence variants.
Maternal intake of vitamin D in pregnancy is a potentially modifiable but understudied risk factor for the development of asthma in children.
We investigated whether maternal vitamin D intake in ...pregnancy is associated with decreased risks of wheezing symptoms in young children.
Subjects were from a birth cohort recruited in utero with the primary objective of identifying associations between maternal diet during pregnancy and asthma and allergies in children. A random sample of 2000 healthy pregnant women was recruited while attending antenatal clinics at the Aberdeen Maternity Hospital, Scotland, at approximately 12 wk gestation. Maternal vitamin D intake was ascertained from a food-frequency questionnaire completed at 32 wk of gestation. The main outcome measures were wheezing symptoms, spirometry, bronchodilator response, atopic sensitization, and exhaled nitric oxide at 5 y.
Respiratory details through 5 y and maternal food-frequency-questionnaire data were available for 1212 children. In models adjusted for potential confounders, including the children's vitamin D intake, a comparison of the highest and lowest quintiles of maternal total vitamin D intake conferred lower risks for ever wheeze odds ratio (OR): 0.48; 95% CI: 0.25, 0.91, wheeze in the previous year (OR: 0.35; 95% CI: 0.15, 0.83), and persistent wheeze (OR: 0.33; 95% CI: 0.11, 0.98) in 5-y-old children. In addition, lower maternal total vitamin D intakes in pregnancy were also associated with decreased bronchodilator response (P = 0.04). No associations were observed between maternal vitamin D intakes and spirometry or exhaled nitric oxide concentrations.
Increasing maternal vitamin D intakes during pregnancy may decrease the risk of wheeze symptoms in early childhood.
The state of computer network technologies has continually advanced at a rapid pace. Software Defined Networking (SDN) and Blockchain (BC) have emerged as complementary technologies providing support ...that facilitates greater security and greater network performance for many domains of application, including the Internet of Things (IoT) ecosystem, ideally resulting in an improvement in our collective quality of life. The proliferation of IoT devices, driven by a wide variety of use cases and its ubiquitous availability, combined with the emergence of SDN and BC, presents rich opportunities for various emerging research efforts. This paper presents a comprehensive survey of the studies in which BC and SDN have been integrated into the IoT ecosystem, referred to hereafter as BC-enabled Software- Defined IoT (BC-SDIoT). First, we discuss the motivations and drivers for integrating BC-enabled SDN and BC-SDIoT, as well as the benefits and drawbacks. Second, we categorize the relevant studies according to six key implementation objectives and ideas that combine BC, SDN, and IoT technologies to create smart, secure, and effective frameworks: Security, computing paradigms (edge and fog computing), trust management, access control & authentication, privacy, and networking. In the corresponding sections, we present the categories (i.e., problem domains) of the aforementioned novel taxonomy and discuss related studies (i.e., solutions) in depth. Finally, we outline potential major challenges, open issues, and future prospects that require further research attention and intensive endeavors for complete and ground-breaking frameworks to broaden newer research domains in BC-SDIoT. This survey paper may be a fruitful primer for a reader investigating the exploitation of BC in SDN and IoT ecosystems.
Zero-mode waveguides provide a powerful technology for studying single-molecule real-time dynamics of biological systems at physiological ligand concentrations. We customized a commercial zero-mode ...waveguide-based DNA sequencer for use as a versatile instrument for single-molecule fluorescence detection and showed that the system provides long fluorophore lifetimes with good signal to noise and low spectral cross-talk. We then used a ribosomal translation assay to show real-time fluidic delivery during data acquisition, showing it is possible to follow the conformation and composition of thousands of single biomolecules simultaneously through four spectral channels. This instrument allows high-throughput multiplexed dynamics of single-molecule biological processes over long timescales. The instrumentation presented here has broad applications to single-molecule studies of biological systems and is easily accessible to the biophysical community.
Hospital one was in an urban region, and serves a larger and more deprived population than hospital two (which was located in a rural area). Table 1 Summary of findings Hospital one Jan 2013–Dec 2022 ...Hospital two July 2015–Dec 2022 Urgent admissions Urgent zero day admissions % zero day to all urgent admissions Urgent admissions Urgent zero day admissions % zero day to all urgent admissions Trend before RRC 0.047 (0.029, 0.065)* 0.024 (0.016, 0.032)* 0.0008 (0.0001, 0.0015)† 0.014 (0.003, 0.024)† 0.010 (0.003, 0.17)† 0.0004 (0.0001, 0.0007)† Trend after RRC 0.037 (−0.055, 0.129)‡ −0.004 (−0.041, 0.033)‡ −0.003 (−0.005, –0.0001)† −0.019 (−0.072, 0.034)‡ −0.005 (−0.020, 0.001)‡ 0.0023 (−0.0116, 0.0162)‡ Trend after RRC relative to trend before RRC −0.010 (−0.104, 0.084)‡ 0.029 (−0.067, 0.009)‡ −0.0036 (−0.0056, –0.0016)* −0.246 (−0.382, –0.110)* −0.153 (−0.206, –0.099)* 0.0066 (−0.0498, 0.0631)‡ Consultant on-call rota 1 in 10 1 in 7 Number of children <16 years 118 165 23 592 Number of children per square mile 134 10 Median deprivation quintile (IQR, 1=most deprived) 2 (1–4) 3 (2–4) Mean distance to hospital (km) 12 32 Number of GPs/10 000 population 4.9 6.4 Zero day admission is an urgent admission where the child is admitted and discharged on the same day. When the number of admissions/1000/month was compared before and after the RRC started, there were negative changes in the number of urgent and zero day admissions after RRC started in hospital two and a negative change in the percentage of zero day/all admissions after RRC started in hospital one (table 1).
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