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Several studies have demonstrated a high incidence of autistic spectrum features in individuals with Smith-Lemli-Opitz syndrome (SLOS). However, do these findings imply a converse relationship that has diagnostic utility? Is SLOS testing implicated when autism spectrum disorder (ASD) is the only clinical indication? AIM To determine if there is any correlation with a clinical indication of ASD and a biochemical diagnosis of SLOS, based on historical test request and assay data. METHODS Six years (2008-2013) of clinical test requests for 7-dehydrocholesterol (7-DHC) level were classified and summarised according to indication and final test result. RESULTS From the audit period, 988 valid test results from post-natal samples were identified. In plasma/serum, mean 7-DHC level was 264.7 μmol/L (normal range

Massively parallel sequencing has markedly improved mendelian diagnostic rates. This study assessed the effects of custom alterations to a diagnostic genomic bioinformatic pipeline in response to clinical need and derived practice recommendations relative to diagnostic rates and efficiency. The Genomic Annotation and Interpretation Application (GAIA) bioinformatics pipeline was designed to detect panel, exome, and genome sample integrity and prioritize gene variants in mendelian disorders. Reanalysis of selected negative cases was performed after improvements to the pipeline. GAIA improvements and their effect on sensitivity are described, including addition of a PubMed search for gene-disease associations not in the Online Mendelian Inheritance of Man database, inclusion of a process for calling low-quality variants (known as QPatch), and gene symbol nomenclature consistency checking. The new pipeline increased the diagnostic rate and reduced staff costs, resulting in a saving of US$844.34 per additional diagnosis. Recommendations for genomic analysis pipeline requirements are summarized. Clinically responsive bioinformatics pipeline improvements increase diagnostic sensitivity and increase cost-effectiveness.

The cause of neurodegeneration in MPS mouse models is the focus of much debate and what the underlying cause of disease pathology in MPS mice is. The timing of development of pathology and when this can be reversed or impacted is the key to developing suitable therapies in MPS. This study is the first of its kind to correlate the biochemical changes with the functional outcome as assessed using non-invasive behaviour testing across multiple mucopolysaccharidosis (MPS) mouse models. In the MPS brain, the primary lysosomal enzyme dysfunction leads to accumulation of primary glycosaminoglycans (GAGs) with gangliosides (G(M2) and G(M3)) being the major secondary storage products. With a focus on the neuropathology, a time course experiment was conducted in MPS I, MPS IIIA, MPS VII (severe and attenuated models) in order to understand the relative timing and level of GAG and ganglioside accumulation and how this correlates to behaviour deficits. Time course analysis from 1 to 6 months of age was conducted on brain samples to assess primary GAG (uronic acid), beta-hexosaminidase enzyme activity and levels of G(M2) and G(M3) gangliosides. This was compared to a battery of non-invasive behaviour tests including open field, inverted grid, rotarod and water cross maze were assessed to determine effects on motor function, activity and learning ability. The results show that the GAG and ganglioside accumulation begins prior to the onset of detectable changes in learning ability and behaviour. Interestingly, the highest levels of GAG and ganglioside accumulation was observed in the MPS IIIA mouse despite having 3% residual enzyme activity. Deficits in motor function were clearly observed in the severe Gus(mps/mps), which were significantly delayed in the attenuated Gus(tm(L175F)Sly) model despite their minimal increase in detectable enzyme activity. This suggests that genotype and residual enzyme activity are not indicative of severity of disease pathology in MPS disease and there exists a window when there are considerable storage products without detectable functional deficits which may allow an alteration to occur with therapy. usc Refereed/Peer-reviewed

Mucopolysaccharidosis type IVA (MPS IVA) is an inborn error of glycosaminoglycan (GAG) catabolism characterized by a deficiency of the lysosomal enzyme, N‐acetylgalactosamine 6‐sulphatase (GALNS). Consequently, partially degraded GAG, chondroitin 6‐sulfate (CS) and keratan sulfate (KS), accumulate in the lysosomes of affected cells, primarily in cartilage resulting in skeletal disease. Excessive urinary excretion of these GAG is often used as the initial biochemical parameter to inform a laboratory diagnosis. Here we present the utility of a CS‐disaccharide with a non‐reducing 6‐sulfated N‐acetylgalactosamine residue (HNAc‐UA (1S))—the enzyme's substrate—for the diagnosis and biochemical monitoring of MPS IVA patients. Following implementation of this method into the diagnostic laboratory, we identified one MPS IVA patient over 3 years of MPS urine screening, with no false positive results from 2050 urines tested. Uniquely, urinary concentrations of HNAc‐UA (1S) are independent of age meaning that age‐related reference ranges are not required. Urinary HNAc‐UA (1S) was also able to identify two MPS IVA siblings who remained misdiagnosed with spondyloepiphyseal dysplasia for 5 years because of normal urinary GAG. HNAc‐UA (1S) could also be used as a biomarker for monitoring response to enzyme replacement therapy (ERT) as there was a drop in urinary concentration following the administration of ERT in all 12 patients and concentrations correlated with urinary KS (R 2 = 0.92). In conclusion, HNAc‐UA (1S) is a reliable, sensitive and specific biomarker for the diagnosis of MPS IVA and can be used to biochemically monitor the response to ERT.

Gaucher disease (GD) is an inherited metabolic disorder characterised by impaired catabolism of the glycosphingolipid, glucosylceramide. The deacetylated derivative, glucosylsphingosine (GluSph, lyso-Gb1) has materialised as a biomarker for GD. Further appraisal of the clinical utility of GluSph is required in terms of its prognostic power to inform disease course and pre-symptomatic testing. In this study, we show that plasma GluSph concentrations are significantly higher in GD patients with neuronopathic disease compared with non-neuronopathic disease, even in the neonatal period. A neonate diagnosed at 1 day of age (homozygous for N370S) due to an affected older sibling, returned GluSph of 70 nmol/L compared with 1070-2620 nmol/L for four neuronopathic patients diagnosed

Expanding treatments for the mucopolysaccharidoses—a family of genetic disorders—place unprecedented demands for accurate, timely diagnosis because best outcomes are seen with early initiation of appropriate therapies. Here we sought to improve the diagnostic odyssey by measuring specific glycosaminoglycan fragments with terminal residues complicit with the genetic defect resulting in precise diagnosis rather than the usual first-line, ambiguous total glycosaminoglycan determinations that return poor diagnostic yield. A derivatizing reagent was added to urine aliquots (0.5 μmol creatinine) before separation of the glycosaminoglycan fragments by liquid chromatography and quantification with electrospray ionization–tandem mass spectrometry using multiple reaction monitoring for 10 targeted fragments plus the internal standard. All 93 mucopolysaccharidosis patients were correctly identified as 1 of 10 subtypes from a total of 723 de-identified subjects—blinded to diagnosis—based on the presence of specific “signature” glycosaminoglycan fragments. Employing reference intervals calculated from 630 unaffected urines, with 99% confidence intervals, provided a laboratory test with 100% specificity and sensitivity. This novel urine assay allows diagnosis of 10 mucopolysaccharidosis subtypes in a single test. The precise quantification of unique glycosaminoglycan fragments also enables longitudinal biochemical monitoring following therapeutic interventions.

A young woman with a history of previously undiagnosed episodic neurological deterioration since early childhood associated with viral illnesses developed fulminant encephalitis. Standard investigations gave no diagnosis. Next-generation sequencing identified a heterozygous mutation c.1754C>T (p.Thr585Met) in the RANBP2 gene, giving the diagnosis of recurrent acute necrotising encephalopathy type 1. This condition is probably underdiagnosed, especially in adults, and should be considered in patients with recurrent encephalopathy.

Maple syrup urine disease is caused by a deficiency of branched-chain alpha-ketoacid dehydrogenase, responsible for degradation of leucine, isoleucine, and valine. Biallelic pathogenic variants in BCKDHA, BCKDHB, or DBT genes result in enzyme deficiency. We report the case of a female infant who presented with mild gross motor delay at 4 months, and seizures with hypoglycaemia at 5 months. Newborn screening returned total leucine/isoleucine at the 99.5th centile of the population; however, as second-tier testing reported minimal alloisoleucine, the results were considered inconsistent with MSUD. Plasma amino acid and urine organic acid analyses at 5 months were, however, consistent with a diagnosis of MSUD. A brain MRI showed bilateral symmetrical T2 hyperintense signal abnormalities involving white matter, globus pallidus, thalamus, brainstem, and dentate nuclei with restricted diffusion. A repeat MRI 10 months post-dietary-intervention showed the resolution of these changes and progression in myelination. Her clinical phenotype, including protein tolerance, correlated with intermediate MSUD. Molecular analysis of all three genes identified two variants of uncertain significance, c.434-15_434-4del and c.365A>G (p. Tyr122Cys) in the DBT gene. The rate of leucine decarboxylation in fibroblasts was reduced, but not to the extent observed in classical MSUD patients, supporting an intermediate form of MSUD. Previously reported mRNA splicing studies supported a deleterious effect of the c.434-15_434-4del variant. This functional evidence and confirmation that the variants were in trans, permitted their reclassification as pathogenic and likely pathogenic, respectively, facilitating subsequent prenatal testing. This report highlights the challenges in identifying intermediate MSUD by newborn screening, reinforcing the importance of functional studies to confirm variant pathogenicity in this era of molecular diagnostics.

In about half of all patients with a suspected monogenic disease, genomic investigations fail to identify the diagnosis. A contributing factor is the difficulty with repetitive regions of the genome, such as those generated by segmental duplications. TheWhole exome, whole genome and long-read DNA sequencing techniques combined with studies of RNA and quantitative proteomics were used to investigate 17 subjects from 16 unrelated families with suspected mitochondrial disease.We report six differentAustralian NHMRC, US Department of Defense, Japanese AMED and JSPS agencies, Australian Genomics Health Alliance and Australian Mito Foundation.

Context 4H or POLR3-related leukodystrophy is an autosomal recessive disorder typically characterized by hypomyelination, hypodontia, and hypogonadotropic hypogonadism, caused by biallelic pathogenic variants in POLR3A, POLR3B, POLR1C, and POLR3K. The endocrine and growth abnormalities associated with this disorder have not been thoroughly investigated to date. Objective To systematically characterize endocrine abnormalities of patients with 4H leukodystrophy. Design An international cross-sectional study was performed on 150 patients with genetically confirmed 4H leukodystrophy between 2015 and 2016. Endocrine and growth abnormalities were evaluated, and neurological and other non-neurological features were reviewed. Potential genotype/phenotype associations were also investigated. Setting This was a multicenter retrospective study using information collected from 3 predominant centers. Patients A total of 150 patients with 4H leukodystrophy and pathogenic variants in POLR3A, POLR3B, or POLR1C were included. Main Outcome Measures Variables used to evaluate endocrine and growth abnormalities included pubertal history, hormone levels (estradiol, testosterone, stimulated LH and FSH, stimulated GH, IGF-I, prolactin, ACTH, cortisol, TSH, and T4), and height and head circumference charts. Results The most common endocrine abnormalities were delayed puberty (57/74; 77% overall, 64% in males, 89% in females) and short stature (57/93; 61%), when evaluated according to physician assessment. Abnormal thyroid function was reported in 22% (13/59) of patients. Conclusions Our results confirm pubertal abnormalities and short stature are the most common endocrine features seen in 4H leukodystrophy. However, we noted that endocrine abnormalities are typically underinvestigated in this patient population. A prospective study is required to formulate evidence-based recommendations for management of the endocrine manifestations of this disorder., Canadian Institutes of Health Research [201610PJT-377869, MOP-G2-341146-159133-BRIDG]; Fondation Les Amis d'Elliot; Leuco-Action; Fondation Lueur d'Espoir pour Ayden; Fondation le Tout pour Loo; Reseau de Medecine Genetique Appliquee of the Fonds de Recherche du Quebec-Sante; Compute Canada; Fonds de Recherche du Quebec en Sante (FRQS) Doctoral Scholarship; Fondation du Grand defi Pierre Lavoie Doctoral Scholarship; McGill Faculty of Medicine F. S.B. Miller Fellowship; Research Institute of the McGill University Health Centre Desjardins Studentship in Child Health Research; Directorate of Higher Education Overseas Scholarship-Dikti Scholarship, Ministry of National Education, Republic of Indonesia; CIHR [201603PJT-148695]; BC Children's Hospital Foundation through its intramural Investigator Grant Award Program (IGAP); National Institute for Neurological Disorders and Stroke [R01NS082094]; Jakob Kamens Chair in Translational Neurotherapeutics; Fonds de Recherche du Quebec-Sante (FRQS); Canadian Institutes of Health Research; European Reference Network for Rare Neurological Disorders (ERN-RND) [739510], This study was supported by grants from the Canadian Institutes of Health Research (201610PJT-377869, MOP-G2-341146-159133-BRIDG), Fondation Les Amis d'Elliot, Leuco-Action, Fondation Lueur d'Espoir pour Ayden, Fondation le Tout pour Loo, and Reseau de Medecine Genetique Appliquee of the Fonds de Recherche du Quebec-Sante to G. Bernard. This research was enabled in part by support provided by Compute Canada (www.computecanada.ca).S.Perrier is supported by the Fonds de Recherche du Quebec en Sante (FRQS) Doctoral Scholarship, the Fondation du Grand defi Pierre Lavoie Doctoral Scholarship, the McGill Faculty of Medicine F. S.B. Miller Fellowship, and the Research Institute of the McGill University Health Centre Desjardins Studentship in Child Health Research. F. K.C. is a recipient of the Directorate of Higher Education Overseas Scholarship-Dikti Scholarship, Ministry of National Education, Republic of Indonesia. W.T. G. received funding from the CIHR (201603PJT-148695) and is supported by the BC Children's Hospital Foundation through its intramural Investigator Grant Award Program (IGAP). B.L. F. was supported by the National Institute for Neurological Disorders and Stroke (R01NS082094). A.V. receives funding from the Jakob Kamens Chair in Translational Neurotherapeutics. G. Bernard has received a Research Scholar Junior 1 award from the Fonds de Recherche du Quebec-Sante (FRQS) (2012-2016) and the New Investigator Salary Award from the Canadian Institutes of Health Research (2017-2022). I.K.M., M. Synofzik, D. Tonduti, B.P.v.d.W., M.S. V.d.K., and N.I.W. are members of the European Reference Network for Rare Neurological Disorders (ERN-RND), project ID 739510.

Summary Background In about half of all patients with a suspected monogenic disease, genomic investigations fail to identify the diagnosis. A contributing factor is the difficulty with repetitive regions of the genome, such as those generated by segmental duplications. The ATAD3 locus is one such region in which recessive deletions and dominant duplications have recently been reported to cause lethal perinatal mitochondrial diseases characterized by pontocerebellar hypoplasia or cardiomyopathy, respectively. Methods Whole-exome, whole-genome, and long-read DNA sequencing techniques combined with studies of RNA and quantitative proteomics were used to investigate 17 subjects from 16 unrelated families with suspected mitochondrial disease. Findings We report 6 different de novo duplications in the ATAD3 gene locus causing a distinctive presentation, including lethal perinatal cardiomyopathy, persistent hyperlactacidemia, and frequently, corneal clouding or cataracts and encephalopathy. The recurrent 68-kb ATAD3 duplications are identifiable from genome and exome sequencing but usually missed by microarrays. The ATAD3 duplications result in the formation of identical chimeric ATAD3A/ATAD3C proteins, altered ATAD3 complexes, and a striking reduction in mitochondrial oxidative phosphorylation complex I and its activity in heart tissue. Conclusions ATAD3 duplications appear to act in a dominant-negative manner and the de novo inheritance infers a low recurrence risk for families, unlike most pediatric mitochondrial diseases. More than 350 genes underlie mitochondrial diseases. In our experience, the ATAD3 locus is now one of the five most common causes of nuclear-encoded pediatric mitochondrial disease, but the repetitive nature of the locus means ATAD3 diagnoses may be frequently missed by current genomic strategies. Funding Australian NHMRC, US Department of Defense, US National Institutes of Health, Japanese AMED and JSPS agencies, Australian Genomics Health Alliance, and Australian Mito Foundation.

CBS deficient individuals undergoing betaine supplementation without sufficient dietary methionine restriction can develop severe hypermethioninemia and brain edema. Brain edema has also been observed in individuals with severe hypermethioninemia without concomitant betaine supplementation. We systematically evaluated reports from 11 published and 4 unpublished patients with CBS deficiency and from additional four cases of encephalopathy in association with elevated methionine. We conclude that, while betaine supplementation does greatly exacerbate methionine accumulation, the primary agent causing brain edema is methionine rather than betaine. Clinical signs of increased intracranial pressure have not been seen in patients with plasma methionine levels below 559 μmol/L but occurred in one patient whose levels did not knowingly exceed 972 μmol/L at the time of manifestation. While levels below 500 μmol/L can be deemed safe it appears that brain edema can develop with plasma methionine levels close to 1000 μmol/L. Patients with CBS deficiency on betaine supplementation need to be regularly monitored for concordance with their dietary plan and for plasma methionine concentrations. Recurrent methionine levels above 500 μmol/L should alert clinicians to check for clinical signs and symptoms of brain edema and review dietary methionine intake. Levels approaching 1000 μmol/L do increase the risk of complications and levels exceeding 1000 μmol/L, despite best dietetic efforts, should be acutely addressed by reducing the prescribed betaine dose.

Fabry disease (FD) results from impaired globotriaosylceramide (Gb3) catabolism, due to a deficiency of the lysosomal hydrolase, α-galactosidase A (α-GalA). As a direct consequence, the deacetylated derivative, globotriaosylsphingosine (lyso-Gb3), is produced and contemporary evidence exemplifies its use as a biomarker. Here we developed a simple method to enable quantification of lyso-Gb3 in just 0.01mL of plasma and explored its concentration in a cohort of 73 Australian FD patients, as well as in individuals with other sphingolipidoses. In 2000 patients without FD, but with related metabolic conditions, lyso-Gb3 returned concentrations of5pmol/mL. In the FD cohort, 53/60 patients with classical mutations returned lyso-Gb3 concentrations≥5pmol/mL whereas only 4/13 patients with "late-onset" mutations had lyso-Gb3≥5pmol/mL. Five females with normal α-GalA activity and genetically confirmed FD returned lyso-Gb3≥5pmol/mL. The prevalence of clinically significant disease including cardiomyopathy, nephropathy and cerebrovascular disease was congruent with higher lyso-Gb3 concentrations. Repeat testing was available for 51 patients-26 undergoing enzyme replacement therapy-and concentrations of lyso-Gb3 remained unaltered throughout 6-18 months independent of sex, mutation or treatment status. Our data suggest that the optimum use of lyso-Gb3 resides in laboratory confirmation of classical FD and for monitoring at least the initial response to therapeutic intervention. There is no evidence that lyso-Gb3 can inform on clinical events.

The vendace (Coregonus albula) is the U.K.’s rarest freshwater fish with only four native populations ever having been recorded. These occur or occurred in two lochs (Castle Loch and Mill Loch) in south-west Scotland and in two lakes (Bassenthwaite Lake and Derwent Water) in north-west England. The two vendace populations in Scotland were extirpated many decades ago, but since 1995 and 1998 those in Bassenthwaite Lake and Derwent Water, respectively, have been monitored by gill netting and hydroacoustics within annual fish community surveys. The population in Bassenthwaite Lake was recorded up to 2000 but not in the following 12 years, while that in Derwent Water has been consistently recorded each year. Likely reasons for the apparent extirpation in Bassenthwaite Lake include the multiple stressors of eutrophication, sedimentation and species introductions including roach (Rutilus rutilus) and ruffe (Gymnocephalus cernuus). Measures are in progress to improve environmental conditions, although lake recovery is anticipated to be slow. In 2013, the fish community survey of Bassenthwaite Lake unexpectedly recorded a single underyearling vendace representing a ‘reappearance’ of this species after an apparent absence of 12 years. In 2014, this was followed by the recording of two adult vendace. There are three possible origins for these individuals. First, vendace may have actually survived locally at very low abundance below the limit of detection and may now be increasing in abundance. Second, the fish may have arrived in Bassenthwaite Lake by moving down the River Derwent from Derwent Water. Third, such downstream movement may have happened some time ago and the individuals are the locally-spawned offspring of such recolonisers. Continued monitoring of the fish community of Bassenthwaite Lake promises to be highly informative for both local conservation management and fundamental coregonid population biology.

We describe a sibling pair displaying an early infantile-onset, progressive neurodegenerative phenotype, with symptoms of developmental delay and epileptic encephalopathy developing from 12 to 14 months of age. Using whole exome sequencing, compound heterozygous variants were identified in SLC5A6, which encodes the sodium-dependent multivitamin transporter (SMVT) protein. SMVT is an important transporter of the B-group vitamins biotin, pantothenate, and lipoate. The protein is ubiquitously expressed and has major roles in vitamin uptake in the digestive system, as well as transport of these vitamins across the blood–brain barrier. Pathogenicity of the identified variants was demonstrated by impaired biotin uptake of mutant SMVT. Identification of this vitamin transporter as the genetic basis of this disorder guided targeted therapeutic intervention, resulting clinically in improvement of the patient’s neurocognitive and neuromotor function. This is the second report of biallelic mutations in SLC5A6 leading to a neurodegenerative disorder due to impaired biotin, pantothenate and lipoate uptake. The genetic and phenotypic overlap of these cases confirms mutations in SLC5A6 as the genetic cause of this disease phenotype. Recognition of the genetic disorder caused by SLC5A6 mutations is essential for early diagnosis and to facilitate timely intervention by triple vitamin (biotin, pantothenate, and lipoate) replacement therapy.

Fifty percent of inborn errors of metabolism are present in later childhood and adulthood, with crises commonly precipitated by minor viral illnesses or increased protein ingestion. Many physicians only consider IEM after more common conditions (such as sepsis) have been considered. In view of the large number of inborn errors, it might appear that their diagnosis requires precise knowledge of a large number of biochemical pathways and their interrelationship. As a matter of fact, an adequate diagnostic approach can be based on the proper use of only a few screening tests. A detailed history of antecedent events, together with these simple screening tests, can be diagnostic, leading to life-saving, targeted treatments for many disorders. Unrecognised, IEM can lead to significant mortality and morbidity. Advice is available 24/7 through the metabolic service based at the major paediatric hospital in each state and Starship Children's Health in New Zealand.

Expanding treatments for the mucopolysaccharidoses-a family of genetic disorders-place unprecedented demands for accurate, timely diagnosis because best outcomes are seen with early initiation of appropriate therapies. Here we sought to improve the diagnostic odyssey by measuring specific glycosaminoglycan fragments with terminal residues complicit with the genetic defect resulting in precise diagnosis rather than the usual first-line, ambiguous total glycosaminoglycan determinations that return poor diagnostic yield.A derivatizing reagent was added to urine aliquots (0.5 μmol creatinine) before separation of the glycosaminoglycan fragments by liquid chromatography and quantification with electrospray ionization-tandem mass spectrometry using multiple reaction monitoring for 10 targeted fragments plus the internal standard.All 93 mucopolysaccharidosis patients were correctly identified as 1 of 10 subtypes from a total of 723 de-identified subjects-blinded to diagnosis-based on the presence of specific "signature" glycosaminoglycan fragments. Employing reference intervals calculated from 630 unaffected urines, with 99% confidence intervals, provided a laboratory test with 100% specificity and sensitivity.This novel urine assay allows diagnosis of 10 mucopolysaccharidosis subtypes in a single test. The precise quantification of unique glycosaminoglycan fragments also enables longitudinal biochemical monitoring following therapeutic interventions.

Short stature is a characteristic feature of most of the mucopolysaccharidoses, a group of inherited lysosomal storage disorders caused by a single enzyme deficiency. MPS patients present with progressive skeletal defects from an early age, including short stature due to impaired cartilage-to-bone conversion (endochondral ossification). The aim of this study was to determine which murine MPS model best reproduces the bone length reduction phenotype of human MPS and use this model to determine the earliest developmental stage when disrupted endochondral ossification first appears. Gusmps/mps mice representing severe MPS VII displayed the greatest reduction in bone elongation and were chosen for histopathological analysis. Tibial development was assessed from E12.5 to 6 months of age. Chondrocytes in the region of the future primary ossification center became hypertrophic at a similar age to normal in the MPS VII mouse fetus, but a delay in bone deposition was observed with an approximate 1 day delay in the formation of the primary ossification centre. Likewise, chondrocytes in the region of the future secondary ossification center also became hypertrophic at the same age as normal in the MPS VII early postnatal mouse. Bone deposition in the secondary ossification centre was delayed by two days in the MPS VII proximal tibia (observed at postnatal day 14 (P14) compared to P12 in normal). The thickness of the tibial growth plate was larger in MPS VII mice from P9 onwards. Abnormal endochondral ossification starts in utero in MPS VII and worsens with age. It is characterized by a normal timeframe for chondrocyte hypertrophy but a delay in the subsequent deposition of bone in both the primary and secondary ossification centres, accompanied by an increase in growth plate thickness. This suggests that the signals for vascular invasion and bone deposition, some of which are derived from hypertrophic chondrocytes, are altered in MPS VII. Refereed/Peer-reviewed

Antenatal screening for fetal anomalies has provided women and their partners with information to make reproductive choices based on the risk of serious chromosomal or structural defects since the 1990s. Alternative tests include first-trimester screening (combined ultrasound and maternal serum markers), second-trimester maternal serum markers and noninvasive cell-free DNA testing. The recent recommendations by the Royal Australian and New Zealand College of Obstetrics and Gynaecology and the Human Genetics Society of Australasia against second-trimester triple testing are based on unsound performance criteria, raise several contestable issues around access and equity and challenge the principles of governments providing affordable options.

The whitefish (Coregonus lavaretus (L.)) exists as only one native population in Wales, U.K., where it occurs in Llyn Tegid and is known as the gwyniad. This population is of national conservation importance but faces a number of environmental threats. Although eutrophication has not directly impacted on gwyniad habitat availability, associated low oxygen levels in the hypolimnion periodically approach its tolerance limit of 2 mg L-1. Anthropogenic water level fluctuations and a 1980 introduction of ruffe (Gymnocephalus cernuus (L.)), a predator of gwyniad eggs, also pose potential threats. Despite these concerns, gill-netting surveys in the 1960s, 1991, 2003 and 2008 have shown no marked changes in gwyniad population biology in terms of length, weight and condition factor. Furthermore, hydroacoustic surveys from 2003 to 2009 have shown an increasing trend in total fish abundance. On the basis of gill-netting data, this increase is shared by gwyniad which in July 2009 reached a population density of 2,302.2 fish ha-1 (lower and upper 95% confidence limits of 1,795.8 and 2,951.4 fish ha-1, respectively). Nevertheless, a precautionary approach has been adopted to the conservation of this unique population and from 2005 to 2007 a translocation programme transferred 81,300 eggs from 366 male and 50 female gwyniad to the nearby water body of Llyn Arenig Fawr. Monitored hatching rates were generally high and in 2009 a hydroacoustic and gill-netting survey revealed that although the fish community of this site was sparse (1.7 fish ha-1, with lower and upper 95% confidence limits of 0.8 and 3.8 fish ha-1), it now included adult gwyniad. Although it does not yet confirm successful translocation, this observation does demonstrate that the gwyniad is able to reach sexual maturity in the recipient site.

Impaired sulfatide catabolism is the primary biochemical insult in patients with the inherited neurodegenerative disease, metachromatic leukodystrophy (MLD), and sulfatide elevation in body fluids is useful in the diagnostic setting. Here we used mass spectrometry to quantify fourteen species of sulfatide, in addition to the deacetylated derivative, lyso-sulfatide, using high pressure liquid chromatography-electrospray ionisation-tandem mass spectrometry in both positive and negative ion mode. A single phase extraction of 0.01 mL of MLD plasma identified all 14 sulfatide species in the positive ion mode but none in the negative ion mode. Interrogation of seven major and seven hydroxylated molecular species, as well as lyso-sulfatide, identified the C18 isoform as the most informative for MLD. The C18 produced a linear response and was below the limit of quantification ( −1 ) in control plasma with concentrations in MLD plasma ranging from 12 to 196 pmol mL −1 . Serial plasma samples from an MLD patient post-therapeutic bone marrow transplant proved similar to non-disease controls with C18 sulfatide concentrations below the limit of quantification, as did samples from three individuals with an arylsulfatase A pseudodeficiency – a population variant which appears deficient upon enzymatic assay, without manifestation of disease. These findings emphasise the utility of the C18 sulfatide species for the diagnosis of MLD and biochemical monitoring of MLD patients. Extension of this approach to a newborn screening card correctly identified an MLD patient at birth with elevated C18 sulfatide at levels almost double that present in the newborn card from his unaffected sibling, suggesting the methodology may have applicability for newborn screening.

In Australia, 58 patients with Gaucher disease were managed by a Gaucher Disease Advisory Committee (GDAC) through a centrally adminis-tered national programme, the Life Savings Drug Program (LSDP). In June 2009, Genzyme Corporation, which manufactures imiglucerase (Cerezyme), the only enzyme replacement therapy (ERT) registered for the treatment of Gaucher disease in Australia at that time, announced that due to a viral contamination problem there would be no further shipments of Cerezyme to Australia prior to the end of 2009. The GDAC allocated available drug supplies in order to maintain treatment to those most in need on a hierarchal clinical severity basis. A cohort of 24 patients with Type 1 Gaucher disease was withdrawn from therapy, 22 of whom had no discernible clinically significant adverse effects when reviewed off therapy for up to 6 months. In this paper, we review the course of 20 of the patients who have been on imiglucerase for periods of at least 24 months after the end of their 'drug holiday'. No patient experienced a bone crisis nor clinical nor magnetic resonance imaging evidence of new avascular necrosis events during this period. Two years after recommencing ERT after a 6-month drug holiday, no patient had developed an overt irreversible complication of their Gaucher disease, with the majority returning to their previous clinical status.

Mucopolysaccharidosis IIIA is a heritable neurodegenerative disorder resulting from the dysfunction of the lysosomal hydrolase sulphamidase. This leads to the primary accumulation of the complex carbohydrate heparan sulphate in a wide range of tissues and the secondary neuronal storage of gangliosides G(M2) and G(M3) in the brain. G(M2) storage is associated with CNS deterioration in the G(M2) gangliosidosis group of lysosomal storage disorders and may also contribute to MPS CNS disease. N-butyldeoxynojirimycin, an inhibitor of ceramide glucosyltransferase activity and therefore of ganglioside synthesis, was administered to MPS IIIA mice both prior to maximal G(M2) and G(M3) accumulation (early treatment) and after the maximum level of ganglioside had accumulated in the brain (late treatment) to determine if behaviour was altered by ganglioside level. Ceramide glucosyltransferase activity was decreased in both treatment groups; however, brain ganglioside levels were only decreased in the late treatment group. Learning in the water cross maze was improved in both groups and the innate fear response was also restored in both groups. A reduction in the expression of inflammatory gene Ccl3 was observed in the early treatment group, while IL1 beta expression was reduced in both treatment groups. Thus, it appears that NB-DNJ elicits a transient decrease in brain ganglioside levels, some modulation of inflammatory cytokines and a functional improvement in behaviour that can be elicited both before and after overt neurological changes manifest. Synopsis: NB-DNJ improves learning and restores the innate fear response in MPS IIIA mice by decreasing ceramide glucosyltransferase activity and transiently reducing ganglioside storage and/or modulating inflammatory signals. (C) 2016 Elsevier Inc. All rights reserved. usc Refereed/Peer-reviewed

Although also formerly present in two lochs in South West Scotland, populations of vendace (Coregonus albula) were until recently believed to persist in only two U.K. lakes, i.e., Bassenthwaite Lake and Derwent Water in North West England. However, although Derwent Water has retained its natural mesotrophic conditions and the status of its vendace population is acceptable, monitoring at Bassenthwaite Lake has failed to record any specimens since 2000 and the species has recently been declared locally extinct. Bassenthwaite Lake has experienced increasing problems from eutrophication, sedimentation and the introduction of fi sh species including roach (Rutilus rutilus) and ruffe (Gymnocephalus cernuus), both of which have more recently also been introduced to Derwent Water and give concern over possible food competition and egg predation. In addition to monitoring, considerable efforts have been made in recent years to conserve vendace through the protection and improvement of their habitats and the establishment of refuge populations. The latter has resulted in the establishment of a population originating from Bassenthwaite Lake in Loch Skeen of South West Scotland, with further attempts still in progress at two other sites. Public awareness of the conservation of this species, which is the U.K.'s rarest freshwater fish, has also been actively promoted.

1. Advancement of phytoplankton peaks has been reported from a large variety of aquatic systems, attributed mostly to climate warming. Most studies have used chlorophyll a as a proxy for phytoplankton biomass. Here, we investigated phenology of three phytoplankton taxa (Asterionella formosa, Aulacoseira spp. and Cryptomonas spp.) in four lakes of the English Lake District situated within the same catchment over a 58-year period (1945–2003). 2. We used two methods (the centre of gravity method and a fitted Weibull-type function) to quantify the timing of the spring bloom. Both methods gave similar results for the diatoms, but not for Cryptomonas spp. 3. The timing of the spring bloom advanced over the study period for A. formosa but was delayed for Aulacoseira spp. and Cryptomonas spp.. These changes were consistent in all four lakes. 4. We tested whether surface water temperatures prior to the time of phytoplankton peak abundance, winter soluble reactive phosphorus (SRP) and silicate concentrations, winter taxon inoculum, Schmidt stability or a combination of these affected phytoplankton phenology in the four lakes. Results were remarkably consistent: each driver either advanced or delayed the peak of each taxon in all four lakes. However, the different drivers affected the taxa differently. 5. Contrary to our expectations, the regional driver, temperature, did not have a consis-tent effect on the three phytoplankton taxa, significantly advancing A. formosa and Aulacoseira spp. in the North Basin of Windermere but significantly delaying Cryptomonas spp. peaks in three lakes. Overall, the locally variable driver, SRP concentration, had a more consistent effect upon phenology, affecting all taxa in nearly all lakes.

1. Pike (Esox lucius) is a key and flexible piscivore in many fresh waters of the northern temperate zone, but no previous studies have provided a continuous long-term perspective on its diet in response to changing environmental conditions. Here, we describe its winter diet from 1976 to 2009 in the North and South Basins of the lake of Windermere, U.K., where climate change, eutrophication and species introductions have combined to induce fundamental changes in the fish community. 2. A total of 6637 adult pike (fork length 390 to 1090 mm) was examined and found to have consumed a total of 4436 fish prey of which 98% of individuals identifiable to species comprised native Arctic charr (Salvelinus alpinus), brown trout (Salmo trutta), perch (Perca fluviatilis) and pike and non-native roach (Rutilus rutilus). Over the 34-year study period, the dietary importance of the salmonids Arctic charr and brown trout decreased, while that of the percid perch, the esocid pike and particularly the cyprinid roach increased. These changes were particularly marked in the more eutrophicated South Basin of the lake. 3. The above chronological trends in species-specific contributions to the diet composition of pike had considerable overall impacts. In the 1970s, pike diet composition was dominated by Arctic charr and brown trout which together comprised 94% of the diet. In contrast, in the 2000s, these two species accounted for just 55% of the diet, with perch and roach now comprising 41%. 4. Recent changes observed in the Windermere fish community of a decrease in native salmonids and an increase in cyprinids are consistent with the generally expected effects of climate change in the northern temperature zone. Here, we have shown that they have led to corresponding changes in the diet composition of pike. In turn, this may have implications for lake’s food web structure through shortening food chain length from the primary producers to the top aquatic predator.

The development of recombinantly manufactured enzyme replacement therapy (ERT) has revolutionised the management of some inherited disorders of metabolism. Gaucher disease was the first lysosomal storage disorder for which ERT became commercially available and ERT remains first-line treatment for affected individuals. In Australia, 70 patients with Gaucher disease are treated through a centrally administered Australian Government national program known as the Life Savings Drug Program (LSDP). Imiglucerase (Cerezyme), manufactured by Genzyme Corporation, is the only ERT currently registered in Australia for the treatment of Gaucher disease. In June 2009, Genzyme Corporation announced the detection of a virus in its Allston Landing manufacturing facility which resulted in inventories of imiglucerase being insufficient to meet projected global demand. The Australian Government sought advice from its Gaucher Disease Advisory Committee (GDAC) on recalculating patient doses in order to ration available imiglucerase to those most in need on a clinical severity basis. Management of this rationing process was urgent and required extensive investigation to develop a clinical severity hierarchy, to review available imiglucerase stock spread across multiple pharmacies, to implement a strategy for redistributing available imiglucerase according to specific patients' recalculated doses, to advise treating doctors and patients concerning these changed circumstances and to consider new monitoring schedules during the drug shortage phase. A cohort of 24 patients was withdrawn from therapy, 22 of whom had no discernable clinical adverse effect. This experience suggests that short-term studies of maintenance therapy without a no-treatment arm may lead to erroneous conclusions and that some patients may have treatment holidays or delayed infusions without short-term adverse outcomes.

MPS IIIA is a lysosomal storage disorder caused by mutations in the sulphamidase gene, resulting in the accumulation of heparan sulphate glycosaminoglycans (HS GAGs). Symptoms predominantly manifest in the CNS and there is no current therapy that effectively addresses neuropathology in MPS IIIA patients. Recent studies in MPS IIIA mice have shown that rhodamine B substrate deprivation therapy (SDT) (also termed substrate reduction therapy/SRT) inhibits GAG biosynthesis and, improves both somatic and CNS disease pathology. Acute overexposure to high doses of rhodamine B results in liver toxicity and is detrimental to reproductive ability. However, the long-term effects of decreasing GAG synthesis, at the low dose sufficient to alter neurological function are unknown. A trans-generational study was therefore initiated to evaluate the continuous exposure of rhodamine B treatment in MPS IIIA mice over 4 generations, including treatment during pregnancy. No alterations in litter size, liver histology or liver function were observed. Overall, there are no long-term issues with the administration of rhodamine B at the low dose tested and no adverse effects were noted during pregnancy in mice.

An invasive species is defined as an alien (or introduced or non-native) species whose establishment and spread threaten ecosystems, habitats or species with harm. Such threats to UK lake fish communities have long been appreciated and this review assembles case histories, including new data, from the largest lakes of Scotland, Northern Ireland, Wales and England to examine the hypothesis that at least some of these introductions have become invasive. Loch Lomond in Scotland has experienced six introductions [chub (Leuciscus cephalus), common bream (Abramis brama), crucian carp (Carassius carassius), dace (Leuciscus leuciscus), gudgeon (Gobio gobio) and ruffe (Gymnocephalus cernuus)], of which the most significant has been that of the percid ruffe, which has been implicated in a recent decline of the native coregonid whitefish (Coregonus lavaretus). In Northern Ireland, the introduction of the cyprinid roach (Rutilus rutilus) to Lough Neagh has apparently had a negative impact on some overwintering waterfowl, although the native coregonid pollan (Coregonus autumnalis) remains abundant. Llyn Tegid in Wales has received three introductions [rudd (Scardinius erythrophthalmus), ruffe and silver bream (Blicca bjoerkna)], although no impacts on the native whitefish or other fish populations have been observed. In England, individuals of at least 12 native and non-native fish species have been brought to Windermere for the purpose of live-baiting, although only those of the cyprinids roach and common bream have established abundant populations. At the same time, the native salmonid Arctic charr (Salvelinus alpinus) has declined markedly while the native esocid pike (Esox lucius) has shown changes in abundance, distribution and individual condition, although these developments have not been shown to be causally linked. None of these introductions were sanctioned by appropriate fisheries or other regulatory bodies and almost all of them probably arose from the release or escape of live-bait used by pike anglers. Of the 10 species introductions documented here, four (common bream, gudgeon, roach and ruffe) have established abundant populations and two of these (roach and ruffe) have apparently caused or currently threaten harm, supporting the hypothesis that at least some of these introductions have become invasive.

IntroductionFabry disease is an X-linked disorder of glycosphingolipid metabolism arising from a deficiency of the enzyme, α-galactosidase A (AGA). In the absence of a family history, biochemical diagnosis is conferred by reduced activity in dried filter paper blood spots (DBS), elevations in globotriaosylsphingosine (lyso-Gb3) and/or identification of a mutation within the GLA gene.MethodsFrom 2008 to 2012, AGA determinations were performed on leucocytes and in 2012, DBS testing was introduced. A mass spectrometric method to quantify lyso-Gb3 in plasma was added to our repertoire of testing in 2016. Mutation analysis was performed using either Sanger or Next Generation Sequencing, the latter confirmed by sequencing.ResultsBetween 2008 and 2012, approximately 1000 samples were screened for Fabry disease using AGA determinations in leucocytes and 29 patients were confirmed positive (diagnosis rate of 2.9%). Since 2012, approximately 4000 samples have been screened from DBS, with 126 patients being confirmed positive (diagnosis rate of 3.2%). There were 24 patients with AGA deficiency and a GLA gene variant which did not have elevated lyso-Gb3 (approximately 20% of positives).DiscussionFabry disease is a relatively complicated disease to diagnose and requires significant interpretation of results from all three tests. As an X-linked disease, classically affected males are easier to identify, but females are more difficult to diagnose as their DBS can be normal, even when symptomatic. Further testing via lyso-Gb3 and mutation analysis is required to make a final biochemical diagnosis. The ease of submitting a DBS for testing has contributed to an increase in test requests.

IntroductionPompe disease is classified by age of onset and presentation. Late-onset Pompe disease (LOPD) generally presents with progressive limb-girdle and respiratory muscle weakness. Diagnosis is confirmed following dried blood spot (DBS) enzyme assay by a second method, either mutation or glucose tetra-saccharide analysis.MethodsFrom 2000 to 2016, acid α –glucosidase (GAA) activity in DBS was measured using the immuno-capture assay of . Umapathysivam et al. (2001) From 2016 testing used the multiplex tandem mass spectrometry method (6-PLEX LC-MSMS, Perkin Elmer), measuring six lysosomal storage disease enzymes from one DBS. Glucose tetra-saccharides were measured by mass spectrometry and mutation analysis of the GAA gene was performed using either Sanger or Next Generation Sequencing, the latter confirmed by sequencing.ResultsBetween 2000 and 2016, approximately 1500 samples were screened for Pompe disease with 77 positives, a diagnosis rate of 5.1%. Of these, 33 were infantile Pompe disease, with 44 LOPD. Since May 2016, a further 850 patients have been screened for Pompe disease using the MSMS method and of these, 28 patients have screened as positive, with 19 confirmed as having Pompe disease, either by molecular testing or urine tetra-saccharide measurement, a diagnosis rate of 2.2%. Of these 19 patients, 17 are LOPD, with only two infants being diagnosed in this time. Seven adults are still to be confirmed by a second test.ConclusionThe number of patients presenting for testing follows the recent Australian government funding for enzyme replacement therapy for adults with LOPD. The ease of submitting a dried blood spot sample for testing has contributed to an increase in test requests, although this may also reflect an increased recognition of this condition. These patients may have already been diagnosed clinically, but in the absence of a treatment, there was little benefit in a formal diagnosis.

Aim To assess the natural history and impact of the secondary bone disease observed in patients with Mucolipidosis II and III.

The UK lies towards the southernmost distribution limit of the circumpolar Arctic charr (Salvelinus alpinus), and native populations of high national conservation value occur in all four component countries. However, given the temperature requirements of this species and the already relatively mild UK climate, there is strong reason to expect a significant negative impact of climate change on local populations. Data were assembled from repeated Arctic charr population assessments using combinations of hydroacoustics, gill netting, entrapment records and fisheries catches at five Scottish populations, five English populations and one Welsh population. These data were then used to test the hypotheses that there has been a widespread decline of Arctic charr in the UK and that it can be attributed at least in part to climate change. Ten of the 11 studied populations exhibited significant 1990-onwards declines in abundance, while only the most northerly population showed a significant increase. Overall, there was a significant positive relationship between the observed population decline ranking and a vulnerability to climate change ranking based on water body latitude, altitude and mean depth. These observations support the hypothesis that this species has suffered a recent and widespread decline in the UK, and although additional factors are undoubtedly also involved in some specific cases, climate change is a significant factor.

The form of Darwinian selection has important ecological and management implications. Negative effects of harvesting are often ascribed to size truncation (i.e. strictly directional selection against large individuals) and resultant decrease in trait variability, which depresses capacity to buffer environmental change, hinders evolutionary rebound and ultimately impairs population recovery. However, the exact form of harvest-induced selection is generally unknown and the effects of harvest on trait variability remain unexplored. Here we use unique data from the Windermere (UK) long-term ecological experiment to show in a top predator (pike, Esox lucius ) that the fishery does not induce size truncation but disruptive (diversifying) selection, and does not decrease but rather increases variability in pike somatic growth rate and size at age. This result is supported by complementary modelling approaches removing the effects of catch selectivity, selection prior to the catch and environmental variation. Therefore, fishing most likely increased genetic variability for somatic growth in pike and presumably favoured an observed rapid evolutionary rebound after fishery relaxation. Inference about the mechanisms through which harvesting negatively affects population numbers and recovery should systematically be based on a measure of the exact form of selection. From a management perspective, disruptive harvesting necessitates combining a preservation of large individuals with moderate exploitation rates, and thus provides a comprehensive tool for sustainable exploitation of natural resources.

OBJECTIVE: Tandem mass spectrometry is widely applied to routine newborn screening but there are no long-term studies of outcome. We studied the clinical outcome at six years of age in Australia. METHODS: In a cohort study, we analyzed the outcome at 6 years for patients detected by screening or by clinical diagnosis among >2 million infants born from 1994 to 1998 (1 017 800, all unscreened) and 1998 to 2002 (461 500 screened, 533 400 unscreened) recording intellectual and physical condition, school placement, other medical problems, growth, treatment, diet, and hospital admissions. Results were analyzed separately for medium-chain acyl-CoA dehydrogenase deficiency (MCADD) and other disorders, and grouped patients as those who presented clinically or died in the first 5 days of life; patients presented later or diagnosed by screening, and those with substantially benign disorders. RESULTS: Inborn errors, excluding phenylketonuria, were diagnosed in 116 of 1 551 200 unscreened infants (7.5/100 000 births) and 70 of 461 500 screened infants (15.2/100 000 births). Excluding MCADD, 21 unscreened patients with metabolic disorders diagnosed after 5 days of life died or had a significant intellectual or physical handicap (1.35/100 000 population) compared with 2 of the screened cohort (0.43/100 000; odds ratio: 3.1 [95% CI: 0.73–13.32]). Considering the likely morbidity or mortality among the expected number of never-diagnosed unscreened patients, there would be a significant difference. Growth distribution was normal in all cohorts. CONCLUSION: Screening by tandem mass spectrometry provides a better outcome for patients at 6 years of age, with fewer deaths and fewer clinically significant disabilities.

A diverse range of sampling techniques has been developed for the assessment of fish populations in lakes and reservoirs around the world, none of which is suitable for all types of fish in all types of standing water habitat. Consequently, a common contemporary approach is the combination of several sampling techniques depending on the habitats and species under study, logistical issues and, in some cases, ethical issues. In accordance with this approach, a protocol for the assessment of Arctic charr (Salvelinus alpinus) populations has been developed to meet the assessment requirements of the European Union (EU) Habitats Directive. The protocol makes extensive use of hydroacoustics (to determine population density by area, hereafter referred to as abundance) and more limited survey gill netting (to determine other biological characteristics while minimising mortalities) to sample this salmonid inhabitant of large and often deep lakes and reservoirs. The protocol is applied for the first time here for Arctic charr in five effectively unfished Scottish lakes and reservoirs (Loch Builg, Loch Doon, Lock Eck, Loch Insh and Loch of Girlsta) between July 2003 and June 2005. Geometric mean abundances of fish of all species ranged from 5.5 fish ha−1 (lower and upper 95% confidence limits of 1.9–15.8 fish ha−1) to 476.9 fish ha−1 (366.5–620.6 fish ha−1), with Arctic charr varying between 1.6 fish ha−1 (0.7–3.6 fish ha−1) and 457.8 fish ha−1 (351.8–595.8 fish ha−1). A total of 405 fish of seven species, i.e. Arctic charr, brown trout (Salmo trutta), eel (Anguilla anguilla), minnow (Phoxinus phoxinus), perch (Perca fluviatilis), whitefish (Coregonus lavaretus) and three-spined stickleback (Gasterosteus aculeatus), was recorded from the five sites. Arctic charr dominated the overall fish communities at three sites, were important at a fourth, but were very rare at a fifth. Age–frequency distributions of Arctic charr encompassed individuals between 1 and 10 years old, although most sites did not show this full range. Assessment outcomes for this diverse group of water bodies ranged from three of favourable status, through one of unfavourable but maintained status, to one of unfavourable and declining status. Following this successful first practical application, comments are made on the protocol in the light of its performance in the field and subsequent data analysis. Wider implications for other assessment activities are also considered, including sampling standardisation and the more demanding community-based, rather than simply population-based, EU Water Framework Directive.

Hyperammonaemic encephalopathy has been infrequently reported after both standard and high-dose chemotherapy for solid and haematological malignancies. It is important to consider this diagnosis for patients with unexplained behaviour changes or encephalopathy and this case report emphasizes this condition and the importance of early diagnosis and is the first reported in association with rituximab-containing chemotherapy.

Climate change is anticipated to have differential effects on the salmonid and percid-cyprinid components of lake fish communities, which may in turn impact the foraging conditions and so population biology of top aquatic predators including northern pike (Esox lucius Linnaeus). Since the 1940s, extensive monitoring has been undertaken on the northern pike, the salmonid Arctic charr (Salvelinus alpinus) and the percid Eurasian perch (Perca fluviatilis) populations and environmental features of the mesotrophic north and eutrophic south basins of Windermere in the English Lake District, UK Analysis of data from 1982 to 2006 revealed basin-specific changes in the degree of eutrophication, but a common warming of the lake in the 1990s was accompanied by contrasting trends in the two basins’ northern pike populations. Following a common temporary increase in catch-per-unit-effort (CPUE) during the 1980s in response to a lake-wide decrease in fishing pressure, CPUE of the south basin subsequently decreased while that of the north basin showed a second increase in the late 1990s to reach a peak in 2000. In addition, the mean weight at length of female northern pike was initially similar between the two basins but then increased in the north basin from the early 1990s to the early 2000s before decreasing, while in the south basin it has shown a decrease since 2000. These changes are interpreted with respect to contrasting trends in potential prey populations, including an overall decrease in the abundance of the coldwater Arctic charr, particularly in the more eutrophic south basin where hypolimnetic oxygen levels have fallen to very low levels, and a recent marked increase in the abundance of the warmwater roach (Rutilus rutilus) in both basins. It is concluded that although indirect in nature, the recent warming of Windermere has had a significant impact on its northern pike populations.

Quantifying the effects of density-dependent and density-independent factors in demographic and dispersal processes remains a major challenge in population ecology. Based on unique long-term capture–mark–recapture (CMR) data (1949–2000) on pike (Esox lucius) from Windermere, United Kingdom, we provide estimates of density-dependent and density-independent effects, under the influence of individual size and sex, on natural survival, fishing mortality, and dispersal. Because survival is expected to be related to the individual growth process, we also explore the degree of parallelism between the two processes by applying the best-supported survival model structure to individual growth data. The CMR data were analyzed using sex- and age-structured multistate models (two lake basins: north and south) assuming no seasonal variation in survival and dispersal. Total survival and dispersal probabilities were insensitive to this assumption, and capture probability was shown to be robust to assumptions about intra-...

In humans, complex I of the respiratory chain is composed of seven mitochondrial DNA (mtDNA)-encoded and 38 nuclear-encoded subunits that assemble together in a process that is poorly defined. To date, only two complex I assembly factors have been identified and how each functions is not clear. Here, we show that the human complex I assembly factor CIA30 (complex I intermediate associated protein) associates with newly translated mtDNA-encoded complex I subunits at early stages in their assembly before dissociating at a later stage. Using antibodies we identified a CIA30-deficient patient who presented with cardioencephalomyopathy and reduced levels and activity of complex I. Genetic analysis revealed the patient had mutations in both alleles of the NDUFAF1 gene that encodes CIA30. Complex I assembly in patient cells was defective at early stages with subunits being degraded. Complementing the deficiency in patient fibroblasts with normal CIA30 using a novel lentiviral system restored steady-state complex I levels. Our results indicate that CIA30 is a crucial component in the early assembly of complex I and mutations in its gene can cause mitochondrial disease.

The north and south basins of Windermere in the English Lake District, UK, support autumn- and spring-spawning populations of Arctic charr, Salvelinus alpinus, which have been studied since the 1930s. Continuous investigations of the population dynamics of Arctic charr at this lake have involved gill netting since 1939, collection of fishery catch-per-unit-effort data since 1966, and hydroacoustic surveys since 1990. Analysis of these and associated long-term data on the abiotic environment and other components of the fish communities revealed recently contrasting fortunes of the Arctic charr populations of the north and south basins, the latter of which has been significantly impacted by eutrophication while both basins have shown elevated water temperatures and increasing roach, Rutilus rutilus, populations. Despite the introduction of phosphate stripping in 1992 and some subsequent initial improvement, the hypolimnion of the south basin still remains significantly eutrophicated and the fishery catch-per-unit-effort in this basin is now at record low levels. In addition, the spatial distribution of roach has expanded to form significant components of the fish communities of inshore and offshore surface habitats, where this cyprinid may compete with Arctic charr for zooplanktonic prey. It is concluded that the Arctic charr populations of Windermere, particularly those of the south basin, currently face a number of significant environmental pressures and continued management action is required to ensure their survival.

The Fabry Registry is a global observational research platform established to define outcome data on the natural and treated course of this rare disorder. Participating physicians submit structured longitudinal data to a centralized, confidential database. This report describes the baseline demographic and clinical characteristics of the first 1765 patients (54% males (16% aged < 20 years) and 46% females (13% < 20 years)) enrolled in the Fabry Registry. The median ages at symptom onset and diagnosis were 9 and 23 years (males) and 13 and 32 years (females), respectively, indicating diagnostic delays in both sexes. Frequent presenting symptoms in males included neurological pain (62%), skin signs (31%), gastroenterological symptoms (19%), renal signs (unspecified) (17%), and ophthalmological signs (11%). First symptoms in females included neurological pain (41%), gastroenterological symptoms (13%), ophthalmological (12%), and skin signs (12%). For those patients reporting renal progression, the median age at occurrence was 38 years for both sexes, but onset of cerebrovascular and cardiovascular events was later in females (median 43 and 47 years, respectively) than in males (38 and 41 years, respectively). This paper demonstrates that in spite of the considerable burden of disease in both sexes that begins to manifest in childhood or adolescence, the recognition of the underlying diagnosis is delayed by 14 years in males and 19 years in females. The Fabry Registry provides data that can increase awareness of common symptoms in all age groups, as well as insight into treated and untreated disease course, leading to improved recognition and earlier treatment, and possibly to improved outcomes for affected individuals.