Genetic and phenotypic characterization of NKX6‐2‐related spastic ataxia and hypomyelination

Background and purpose Hypomyelinating leukodystrophies are a heterogeneous group of genetic disorders with a wide spectrum of phenotypes and a high rate of genetically unsolved cases. Bi‐allelic mutations in NKX6‐2 were recently linked to spastic ataxia 8 with hypomyelinating leukodystrophy. Methods Using a combination of homozygosity mapping, exome sequencing, and detailed clinical and neuroimaging assessment a series of new NKX6‐2 mutations in a multicentre setting is described. Then, all reported NKX6‐2 mutations and those identified in this study were combined and an in‐depth analysis of NKX6‐2‐related disease spectrum was provided. Results Eleven new cases from eight families of different ethnic backgrounds carrying compound heterozygous and homozygous pathogenic variants in NKX6‐2 were identified, evidencing a high NKX6‐2 mutation burden in the hypomyelinating leukodystrophy disease spectrum. Our data reveal a phenotype spectrum with neonatal onset, global psychomotor delay and worse prognosis at the severe end and a childhood onset with mainly motor phenotype at the milder end. The phenotypic and neuroimaging expression in NKX6‐2 is described and it is shown that phenotypes with epilepsy in the absence of overt hypomyelination and diffuse hypomyelination without seizures can occur. Conclusions NKX6‐2 mutations should be considered in patients with autosomal recessive, very early onset of nystagmus, cerebellar ataxia with hypotonia that rapidly progresses to spasticity, particularly when associated with neuroimaging signs of hypomyelination. Therefore, it is recommended that NXK6‐2 should be included in hypomyelinating leukodystrophy and spastic ataxia diagnostic panels

Biallelic mutations in SORD cause a common and potentially treatable hereditary neuropathy with implications for diabetes

Here we report biallelic mutations in the sorbitol dehydrogenase gene (SORD) as the most frequent recessive form of hereditary neuropathy. We identified 45 individuals from 38 families across multiple ancestries carrying the nonsense c.757delG (p.Ala253GlnfsTer27) variant in SORD, in either a homozygous or compound heterozygous state. SORD is an enzyme that converts sorbitol into fructose in the two-step polyol pathway previously implicated in diabetic neuropathy. In patient-derived fibroblasts, we found a complete loss of SORD protein and increased intracellular sorbitol. Furthermore, the serum fasting sorbitol levels in patients were dramatically increased. In Drosophila, loss of SORD orthologs caused synaptic degeneration and progressive motor impairment. Reducing the polyol influx by treatment with aldose reductase inhibitors normalized intracellular sorbitol levels in patient-derived fibroblasts and in Drosophila, and also dramatically ameliorated motor and eye phenotypes. Together, these findings establish a novel and potentially treatable cause of neuropathy and may contribute to a better understanding of the pathophysiology of diabetes

PDXK mutations cause polyneuropathy responsive to pyridoxal 5'-phosphate supplementation.

OBJECTIVE: To identify disease-causing variants in autosomal recessive axonal polyneuropathy with optic atrophy and provide targeted replacement therapy. METHODS: We performed genome-wide sequencing, homozygosity mapping, and segregation analysis for novel disease-causing gene discovery. We used circular dichroism to show secondary structure changes and isothermal titration calorimetry to investigate the impact of variants on adenosine triphosphate (ATP) binding. Pathogenicity was further supported by enzymatic assays and mass spectroscopy on recombinant protein, patient-derived fibroblasts, plasma, and erythrocytes. Response to supplementation was measured with clinical validated rating scales, electrophysiology, and biochemical quantification. RESULTS: We identified biallelic mutations in PDXK in 5 individuals from 2 unrelated families with primary axonal polyneuropathy and optic atrophy. The natural history of this disorder suggests that untreated, affected individuals become wheelchair-bound and blind. We identified conformational rearrangement in the mutant enzyme around the ATP-binding pocket. Low PDXK ATP binding resulted in decreased erythrocyte PDXK activity and low pyridoxal 5'-phosphate (PLP) concentrations. We rescued the clinical and biochemical profile with PLP supplementation in 1 family, improvement in power, pain, and fatigue contributing to patients regaining their ability to walk independently during the first year of PLP normalization. INTERPRETATION: We show that mutations in PDXK cause autosomal recessive axonal peripheral polyneuropathy leading to disease via reduced PDXK enzymatic activity and low PLP. We show that the biochemical profile can be rescued with PLP supplementation associated with clinical improvement. As B6 is a cofactor in diverse essential biological pathways, our findings may have direct implications for neuropathies of unknown etiology characterized by reduced PLP levels. ANN NEUROL 2019;86:225-240

The Animal Trypanosomiases and their chemotherapy:a review

Pathogenic animal trypanosomes affecting livestock have represented a major constraint to agricultural development in Africa for centuries, and their negative economic impact is increasing in South America and Asia. Chemotherapy and chemoprophylaxis represent the main means of control. However, research into new trypanocides has remained inadequate for decades, leading to a situation where the few compounds available are losing efficacy due to the emergence of drug-resistant parasites. In this review, we provide a comprehensive overview of the current options available for the treatment and prophylaxis of the animal trypanosomiases, with a special focus on the problem of resistance. The key issues surrounding the main economically important animal trypanosome species and the diseases they cause are also presented. As new investment becomes available to develop improved tools to control the animal trypanosomiases, we stress that efforts should be directed towards a better understanding of the biology of the relevant parasite species and strains, to identify new drug targets and interrogate resistance mechanisms

AMPA receptor GluA2 subunit defects are a cause of neurodevelopmental disorders.

AMPA receptors (AMPARs) are tetrameric ligand-gated channels made up of combinations of GluA1-4 subunits encoded by GRIA1-4 genes. GluA2 has an especially important role because, following post-transcriptional editing at the Q607 site, it renders heteromultimeric AMPARs Ca2+-impermeable, with a linear relationship between current and trans-membrane voltage. Here, we report heterozygous de novo GRIA2 mutations in 28 unrelated patients with intellectual disability (ID) and neurodevelopmental abnormalities including autism spectrum disorder (ASD), Rett syndrome-like features, and seizures or developmental epileptic encephalopathy (DEE). In functional expression studies, mutations lead to a decrease in agonist-evoked current mediated by mutant subunits compared to wild-type channels. When GluA2 subunits are co-expressed with GluA1, most GRIA2 mutations cause a decreased current amplitude and some also affect voltage rectification. Our results show that de-novo variants in GRIA2 can cause neurodevelopmental disorders, complementing evidence that other genetic causes of ID, ASD and DEE also disrupt glutamatergic synaptic transmission

PDXK mutations cause polyneuropathy responsive to PLP supplementation

OBJECTIVE: To identify disease-causing variants in autosomal recessive axonal polyneuropathy with optic atrophy and provide targeted replacement therapy. METHODS: We performed genome-wide sequencing, homozygosity mapping and segregation analysis for novel disease-causing gene discovery. We used circular dichroism to show secondary structure changes and isothermal titration calorimetry to investigate the impact of variants on ATP-binding. Pathogenicity was further supported by enzymatic assays and mass spectroscopy on recombinant protein, patient-derived fibroblasts, plasma and erythrocytes. Response to supplementation was measured with clinical validated rating scales, electrophysiology and biochemical quantification. RESULTS: We identified bi-allelic mutations in PDXK in five individuals from two unrelated families with primary axonal polyneuropathy and optic atrophy. The natural history of this disorder suggests that untreated, affected individuals become wheelchair-bound and blind. We identified conformational rearrangement in the mutant enzyme around the ATP-binding pocket. Low PDXK ATP-binding resulted in decreased erythrocyte PDXK activity and low pyridoxal 5'-phosphate (PLP) concentrations. We rescued the clinical and biochemical profile with PLP supplementation in one family, improvement in power, pain and fatigue contributing to patients regaining their ability to ambulate during the first year of PLP normalization. INTERPRETATION: We show that mutations in PDXK cause autosomal recessive axonal peripheral polyneuropathy leading to disease via reduced PDXK enzymatic activity and low PLP. We show that the biochemical profile can be rescued with PLP supplementation associated with clinical improvement. As B6 is a cofactor in diverse essential biological pathways, our findings may have direct implications for neuropathies of unknown aetiology characterised by reduced PLP levels. This article is protected by copyright. All rights reserved

Solute transport dynamics by high-resolution dye tracer experiments - Image analysis and time moments

Accurate measurements of solute concentration are needed to conduct studies of solute transport process in unsaturated soil. In this paper we present a method of obtaining accurate measurements in and space using dye infiltration and image analysis. The soil color was related to the dye concentration in the soil (C-s) using 74 small calibration samples. The overall root mean square error (RMSE) 0.057 g dm(-3), however, for C-s < 0.75 g dm(-3), the RMSE was only 0.032 g dm(-3). Variability of the concentration estimates at the pixel scale could be reduced by using an average filter. We used the calibration relationship during four infiltration experiments in a 0.95 by 0.975 m large Plexiglas Hele-Shaw cell to calculate dye concentration patterns. Using the first and second order time moments, the dispersivity lambda was calculated for nine different artificial column widths, from 0.0014 (local-scale) to 0.72 m (meso-scale). The horizontally averaged lambda proved to be identical for column widths from 0.0014 to 0.045 m. For larger scales, lambda gradually increased. We noticed that the two experiments with higher flow (1 and 2) and the two experiments with lower flow (3 and 4) showed an almost identical variation of meso-scale lambda with depth. We concluded that above a specific critical value of theta (similar to 0.22 m(3) m(-3)), solute mixing is enhanced, leading to a lower lambda, and that solute transport can be described as a convective-dispersive process. When theta is lower than this critical level, part of the porosity is deactivated and mixing between individual stream tubes decreases, which implies that transport then occurs as a stochastic-convective process

Is there association between hyperdense middle cerebral artery sign on CT scan and time from stroke onset within the first 24-hours?

BACKGROUND: The hyperdense artery sign (HAS) on CT brain scan is an assumed radiological marker of acute intra-arterial thrombotic occlusion. However, the relationship between HAS between time of stroke onset has not been adequately investigated, leading to uncertainty regarding its validity as a marker of acute ischaemia. We attempted to determine if the presence of the hyperdense artery sign is associated with time from stroke onset. METHODS: Retrospective cross-sectional study conducted in a tertiary referral centre. Consecutive patients with acute ischaemic stroke and confirmed middle cerebral arterial occlusion on initial CT angiogram from 2007-2011 were included. Visual estimation and manual measurement of Hounsfield units of affected and corresponding non-affected artery on non-contrast CT was completed and mean density was calculated from four separate readings. Primary outcome measures were Time from stroke onset and HAS on both visual estimation and the ratio of mean value in Hounsfield Units (HU) of affected to non-affected artery. RESULTS: One hundred and fifty-four subjects with confirmed arterial occlusion on CT Angiogram were included in the study. There were no significant differences in age distribution or vascular risk factor presence between subjects with or without HAS. Subjects with HAS were less likely to be male (50.9% vs 70.8%, p = 0.02).) HAS was found in 106 (68.8%) of all subjects. Median NIHSS score at presentation was significantly higher in the HAS group (17 vs 12, p = 0.02). No statistically significant association between HAS and stroke onset time or density ratio between affected and non-affected artery was detected overall within either the first 24-h or on subgroup analysis of those in the first 4.5-h. A small subgroup of three patients with stroke onset greater than 24-h all had absent HAS. CONCLUSIONS: No evidence of a correlation between time of stroke onset and presence of a HAS within the first 24-h post acute ischaemic stroke was identified. The HAS was associated with a higher NIHSS score at presentation

Tobacco and tobacco branding in films most popular in the UK from 2009 to 2017

Background: Exposure to tobacco content in films is a cause of smoking uptake in young people. In an earlier study, we reported that tobacco content occurred in 70% of UK box office films popular between 1989 and 2008. We now report an analysis of tobacco content in a sample of the top grossing UK box office films between 2009 and 2017, and of population exposure resulting from audience exposure to the 2017 films.Methods: Occurrence of tobacco intervals (actual tobacco use, implied use, appearance of smoking paraphernalia or branding) was measured by 5 min interval coding in the 15 most commercially successful films in the UK in each year from 2009 to 2017. A nationally representative survey was used to estimate population exposure to the top 15 films from 2017.Results: We coded 3248 intervals from the 135 films. Tobacco content appeared in 245 intervals (8%, 95% CI 7% to 9%) across 56 (41%, 95% CI 33% to 49%) films. Tobacco content occurred in films in all BBFC age ratings, and 36 (64%, 95% CI 51% to 77%) of films containing tobacco imagery were classified as suitable for viewing by people aged under 15 years. Although less prevalent than in our earlier study, there was no evidence of a secular decline in tobacco content during this study period. The top 15 films from 2017 delivered approximately 21.6 (95% CI 21.06–22.14) million tobacco impressions to young people aged 10–18 years in the UK.Conclusions: Tobacco content continues to appear in UK Box Office films and is widely seen by young people, representing a major driver of smoking uptake

Ontogenetic Change in the Temporal Region of the Early Permian Parareptile <i>Delorhynchus cifellii</i> and the Implications for Closure of the Temporal Fenestra in Amniotes

<div><p>A juvenile specimen of <i>Delorhynchus cifellii</i>, collected from the Early Permian fissure-fill deposits of Richards Spur, Oklahoma, permits the first detailed study of cranial ontogeny in this parareptile. The specimen, consisting of a partially articulated skull and mandible, exhibits several features that identify it as juvenile. The dermal tuberosities that ornament the dorsal side and lateral edges of the largest skull of <i>D</i>. <i>cifellii</i> specimens, are less prominent in the intermediate sized holotype, and are absent in the new specimen. This indicates that the new specimen represents an earlier ontogenetic stage than all previously described members of this species. In addition, the incomplete interdigitation of the sutures, most notably along the fronto-nasal contact, plus the proportionally larger sizes of the orbit and temporal fenestrae further support an early ontogenetic stage for this specimen. Comparisons between this juvenile and previously described specimens reveal that the size and shape of the temporal fenestra in <i>Delorhynchus</i> appear to vary through ontogeny, due to changes in the shape and size of the bordering cranial elements. The jugal of the juvenile specimen is tri-radiate and similar in outline with those found in other amniotes with temporal fenestrae. The available growth series of <i>D</i>. <i>cifellii</i> shows that the jugal gradually becomes a more robust, tetra-radiate element, as the proportionate size of the temporal fenestra is reduced. Ontogenetic changes of other elements that form the border of the fenestra also contribute to its reduction. This growth series provides valuable new information regarding the ontogenetic trajectory of the temporal fenestra in a Palaeozoic reptile, which may be applicable to the evolutionary event of loss of temporal fenestration in other amniotes.</p></div