Potential diagnostic aids for abnormal fatty acid metabolism in a range of neurodevelopmental disorders

Disorders of neurodevelopment include attention deficit hyperactivity disorder, dyspraxia, dyslexia and autism. There is considerable co-morbidity of these disorders and their identification often presents difficulties to those making a diagnosis. This is especially difficult when a multidisciplinary approach is not adopted. All of these disorders have been reported as associated with fatty acid abnormalities ranging from genetic abnormalities in the enzymes involved in phospholipid metabolism to symptoms reportedly improved following dietary supplementation with long chain fatty acids. If definitive disorders of lipid metabolism could be defined then the diagnosis and subsequent management of neurodevelopmental disorders might be transformed. In the identification of those disorders of development which involve lipid metabolism, there are now several tests, measures of lipid metabolism, which could be useful.     

Mouse models of fukutin-related protein mutations show a wide range of disease phenotypes

Dystroglycanopathies are characterized by a reduction in the glycosylation of alpha-dystroglycan (α-DG). A common cause for this subset of muscular dystrophies is mutations in the gene of fukutin-related protein (FKRP). FKRP mutations have been associated with a wide spectrum of clinical severity from severe Walker–Warburg syndrome and muscle–eye–brain disease with brain and eye defects to mild limb–girdle muscular dystrophy 2I with myopathy only. To examine the affects of FKRP mutations on the severity of the disease, we have generated homozygous and compound heterozygous mouse models with human mutations in the murine FKRP gene. P448Lneo+ and E310delneo+ mutations result in severe dystrophic and embryonic lethal phenotypes, respectively. P448Lneo+/E310delneo+ compound heterozygotes exhibit brain defects and severe muscular dystrophies with near absence of α-DG glycosylation. Removal of the Neo^r cassette from the P448Lneo+ homozygous mice eliminates overt brain and eye defects, and reduces severity of dystrophic phenotypes. Furthermore, introduction of the common L276I mutation to generate transgenic L276Ineo+ homozygous and L276Ineo+/P448Lneo+ and L276Ineo+/E310delneo+ compound heterozygotes results in mice displaying milder dystrophies with reduced α-DG glycosylation and no apparent brain defects. Limited sampling and variation in functionally glycosylated α-DG levels between and within muscles may explain the difficulties in correlating FKRP expression levels with phenotype in clinics. The nature of individual mutations, expression levels and status of muscle differentiation all contribute to the phenotypic manifestation. These mutant FKRP mice are useful models for the study of disease mechanism(s) and experimental therapies.     

Prenatal and postnatal animal models of immune activation: Relevance to a range of neurodevelopmental disorders

Epidemiological evidence has established links between immune activation during the prenatal or early postnatal period and increased risk of developing a range of neurodevelopment disorders in later life. Animal models have been used to great effect to explore the ramifications of immune activation during gestation and neonatal life. A range of behavioral, neurochemical, molecular, and structural outcome measures associated with schizophrenia, autism, cerebral palsy, and epilepsy have been assessed in models of prenatal and postnatal immune activation. However, the epidemiology-driven disease-first approach taken by some studies can be limiting and, despite the wealth of data, there is a lack of consensus in the literature as to the specific dose, timing, and nature of the immunogen that results in replicable and reproducible changes related to a single disease phenotype. In this review, we highlight a number of similarities and differences in models of prenatal and postnatal immune activation currently being used to investigate the origins of schizophrenia, autism, cerebral palsy, epilepsy, and Parkinson's disease. However, we describe a lack of synthesis not only between but also within disease-specific models. Our inability to compare the equivalency dose of immunogen used is identified as a significant yet easily remedied problem. We ask whether early life exposure to infection should be described as a disease-specific or general vulnerability factor for neurodevelopmental disorders and discuss the implications that either classification has on the design, strengths and limitations offuture experiments. ? 2012 Wiley Periodicals, Inc. Develop Neurobiol, 2012.     

Recurrent CNVs on 1q21.1, 15q 13.3 and 16p 11.2: shared genetic risk factors for neurodevelopmental and neuropsychiatric disorders

In the last couple of years, the discovery and characterization of copy-number variations （CNVs） in the human genome have provided a new research tool for identifying genetic risk factors of common complex diseases, referred to as CNVs based genome-wide association studies （GWAS）. As ever smaller genomic rearrangements have been discovered by high resolution GWAS, both de novo and inheritable CNVs （also referred to as microdeletion/microduplication or genomic imbalances） have emerged as an important type of genetic risk factor associated with complex diseases. Recent studies with large-scale, multiple cohorts have not only demonstrated the clinical significance of CNVs but also provided evidence suggesting that some CNVs may associate with various neurodevelopmental and neuropsychiatric disorders.     

Nomadism and seasonal range expansion in a large frugivorous bird

Studies on the ranging behaviour of birds often suggest that ranges vary seasonally with larger ranges in the non‐breeding compared to the breeding season. However, due to limitations in tracking methods very little is known about the underlying processes driving seasonal differences in ranging behaviour, especially in fragmented, heterogeneous landscapes. Such knowledge is particularly important if movements deliver essential ecosystem functions such as seed dispersal. We contrasted the daily ranging behaviour between the breeding and non‐breeding season of a frugivorous bird and demonstrate how larger seasonal ranges in the non‐breeding season emerge through switching from a stationary home range behaviour to nomadism. We tracked movements of 29 male trumpeter hornbills Bycanistes bucinator across a fragmented landscape of eastern South Africa during different breeding and non‐breeding seasons using high temporal resolution GPS data‐loggers. Birds in the breeding seasons showed a typical, stationary home range pattern. In the non‐breeding seasons birds, rather than expanding their stationary daily ranges, switched to nomadic movements that were characterized by shifts of the general location of daily ranges to a different area every couple of days. We also found that during the breeding seasons hornbills were mostly located in large continuous forests; birds in the non‐breeding seasons frequently used forest patches within the agricultural landscape and residential areas. These seasonal differences in the movement behaviour of trumpeter hornbills may have important consequences for seed dispersal of plant species. Our findings show how seasonal range expansion of frugivorous birds may be driven by fundamental behavioural changes that have important consequences for ecosystem processes.     

Shear linear behavior of brain tissue over a large frequency range

The literature review about the shear linear properties of brain tissue reveals both a large discrepancy in the existing data and a crucial lack of information at high frequencies associated with traffic road and non-penetrating ballistic impacts. The purpose of this study is to clarify and to complement the linear material characterisation of brain tissue. New data at small strains and high frequencies were obtained from oscillatory experiments. The tests were performed on thin porcine white matter samples (corona radiata) using an original custom-designed oscillatory shear testing device. At 37 degrees C, the results showed that the mean storage modulus (G') and the mean loss modulus (G') increased with the frequency (0.1 to 6310 Hz) from 2.1+/-0.9 kPa to 16.8+/-2.0 kPa and from 0.4+/-0.2 kPa to 18.7+/-2.3 kPa respectively. The reliability of these new dynamic data was checked over a partially common frequency range by conducting similar experiments using a standard rheometer (Bohlin C-VOR 150). Data were also compared in the time field. From these experiments, the relaxation modulus (G(t)) was found to decrease from 24.4+/-2.1 kPa to 1.0+/-0.3 kPa between 10(-5) s and 270 s.     

CNVs: harbingers of a rare variant revolution in psychiatric genetics

The genetic bases of neuropsychiatric disorders are beginning to yield to scientific inquiry. Genome-wide studies of copy number variation (CNV) have given rise to a new understanding of disease etiology, bringing rare variants to the forefront. A proportion of risk for schizophrenia, bipolar disorder, and autism can be explained by rare mutations. Such alleles arise by de novo mutation in the individual or in recent ancestry. Alleles can have specific effects on behavioral and neuroanatomical traits; however, expressivity is variable, particularly for neuropsychiatric phenotypes. Knowledge from CNV studies reflects the nature of rare alleles in general and will serve as a guide as we move forward into a new era of whole-genome sequencing.     

Pattern formation in a ts-cell-lethal mutant ofDrosophila: The range of phenotypes induced by larval heat treatments

Summary We describe the range of phenotypes caused by cell death when larvae of the heat-sensitive cell-lethal mutant,l(1)ts726 ofDrosophila, are subjected to heat treatment at different stages of development. When the treatment extends into the pupal period, certain bristles fail to develop but the disc derivatives are otherwise normal. Earlier treatments cause the replacement of sets of leg and eye-antennal markers by mirror image duplications of neighbouring sets. The results are compared in detail with those expected under a gradient model proposed earlier to account for the phenotype. It is found that although the results for the second leg are in excellent agreement with the predictions of the model, a more elaborate hypothesis is necessary to account for the eye-antennal disc data. Abnormal head patterns fall into several distinct categories, any one of which could be explained by postulating the existence of a gradient, if other categories did not also occur. The markers affected in each case belong to overlapping sets, and each category of pattern can be induced by heat treatments administered throughout the temperature-sensitive period. The statistical distribution of the data is such that only one category of pattern would be detected in a small scale experiment. The possible implications relative to pattern formation in normal development are discussed.Supported by NRC (Canada) grant number A6485 to M.A.R. and funds from the Committee for Computer Utilisation of the University of Alberta     

Nitrogen concentrations in a mangrove creek with a large tidal range,Peninsular Malaysia

We report on various species of nitrogen in subsurface water samples taken at the mouth of Sungai Sementa Besar, a creek about 4.5 km long off the Straits of Malacca and draining a mangrove forest on the west coast of Peninsular Malaysia. This tide-dominated watercourse shows a range greater than 4.5 metres from low to high water during springs, only 1.0 m during neaps, and salinities at the mouth of 30 or greater. After heavy rains or when the tides inundate the forest floor, inorganic nitrogen values increase as much as tenfold at the mouth. Particulate and total organic nitrogen also increase at the mouth under these conditions. There were also comparative highs of organic inputs at other times.     

Overexpression of polygalacturonase in transgenic apple trees leads to a range of novel phenotypes involving changes in cell adhesion

Polygalacturonases (PGs) cleave runs of unesterified GalUA that form homogalacturonan regions along the backbone of pectin. Homogalacturonan-rich pectin is commonly found in the middle lamella region of the wall where two adjacent cells abut and its integrity is important for cell adhesion. Transgenic apple (Malus domestica Borkh. cv Royal Gala) trees were produced that contained additional copies of a fruit-specific apple PG gene under a constitutive promoter. In contrast to previous studies in transgenic tobacco (Nicotiana tabacum) where PG overexpression had no effect on the plant (K.W. Osteryoung, K. Toenjes, B. Hall, V. Winkler, A.B. Bennett [1990] Plant Cell 2: 1239-1248), PG overexpression in transgenic apple led to a range of novel phenotypes. These phenotypes included silvery colored leaves and premature leaf shedding due to reduced cell adhesion in leaf abscission zones. Mature leaves had malformed and malfunctioning stomata that perturbed water relations and contributed to a brittle leaf phenotype. Chemical and ultrastructural analyses were used to relate the phenotypic changes to pectin changes in the leaf cell walls. The modification of apple trees by a single PG gene has offered a new and unexpected perspective on the role of pectin and cell wall adhesion in leaf morphology and stomatal development.     

Novel model to analyze the effect of a large compressive follower pre-load on range of motions in a lumbar spine

A 3-D finite element model (FEM) of the lumbar spine (L1-S1) was used to determine the effect of a large compressive follower pre-load on range of motions (ROM) in all three planes. The follower load modeled in the FEM produced minimal vertebral rotations in all the three planes. The model was validated by comparing the disc compression at all levels in the lumbar spine with the corresponding results obtained by compressing 10 cadevaric lumbar spines (L1-S1) using the follower load technique described by Patwardhan et al. [1999. A follower load increases the load-carrying capacity of the lumbar spine in compression. Spine 24(10), 1003-1009]. Further validation of the model was performed by comparing the lateral bending and torsion response without pre-load and the flexion-extension response without pre-load and with an 800 N follower pre-load with those obtained using cadaver lumbar spines. Following validation, the FEM was subjected to bending moments in all three planes with and without compressive follower pre-loads of up to 1200 N. Disc compression values and the flexion-extension range of motion under 800 N follower pre-load predicted by the FEM compared well with in vitro results. The current model showed that compressive follower pre-load decreased total as well as segmental ROM in flexion-extension by up to 18%, lateral bending by up to 42%, and torsion by up to 26%.     

Major influence of repetitive elements on disease-associated copy number variants (CNVs)

Copy number variants (CNVs) are important contributors to the human pathogenic genetic diversity as demonstrated by a number of cases reported in the literature. The high homology between repetitive elements may guide genomic stability which will give rise to CNVs either by non-allelic homologous recombination (NAHR) or non-homologous end joining (NHEJ). Here, we present a short guide based on previously documented cases of disease-associated CNVs in order to provide a general view on the impact of repeated elements on the stability of the genomic sequence and consequently in the origin of the human pathogenic variome.     

Mutualist‐mediated effects on species' range limits across large geographic scales

Understanding the processes determining species range limits is central to predicting species distributions under climate change. Projected future ranges are extrapolated from distribution models based on climate layers, and few models incorporate the effects of biotic interactions on species' distributions. Here, we show that a positive species interaction ameliorates abiotic stress, and has a profound effect on a species' range limits. Combining field surveys of 92 populations, 10 common garden experiments throughout the range, species distribution models and greenhouse experiments, we show that mutualistic fungal endophytes ameliorate drought stress and broaden the geographic range of their native grass host Bromus laevipes by thousands of square kilometres (~ 20% larger) into drier habitats. Range differentiation between fungal‐associated and fungal‐free grasses was comparable to species‐level range divergence of congeners, indicating large impacts on range limits. Positive biotic interactions may be underappreciated in determining species' ranges and species' responses to future climates across large geographic scales.     

Cas9‐mediated mutagenesis of potato starch‐branching enzymes generates a range of tuber starch phenotypes

We investigated whether Cas9‐mediated mutagenesis of starch‐branching enzymes (SBEs) in tetraploid potatoes could generate tuber starches with a range of distinct properties. Constructs containing the Cas9 gene and sgRNAs targeting SBE1, SBE2 or both genes were introduced by Agrobacterium‐mediated transformation or by PEG‐mediated delivery into protoplasts. Outcomes included lines with mutations in all or only some of the homoeoalleles of SBE genes and lines in which homoeoalleles carried several different mutations. DNA delivery into protoplasts resulted in mutants with no detectable Cas9 gene, suggesting the absence of foreign DNA. Selected mutants with starch granule abnormalities had reductions in tuber SBE1 and/or SBE2 protein that were broadly in line with expectations from genotype analysis. Strong reduction in both SBE isoforms created an extreme starch phenotype, as reported previously for low‐SBE potato tubers. HPLC‐SEC and ¹H NMR revealed a decrease in short amylopectin chains, an increase in long chains and a large reduction in branching frequency relative to wild‐type starch. Mutants with strong reductions in SBE2 protein alone had near‐normal amylopectin chain‐length distributions and only small reductions in branching frequency. However, starch granule initiation was enormously increased: cells contained many granules of <4 μm and granules with multiple hila. Thus, large reductions in both SBEs reduce amylopectin branching during granule growth, whereas reduction in SBE2 alone primarily affects numbers of starch granule initiations. Our results demonstrate that Cas9‐mediated mutagenesis of SBE genes has the potential to generate new, potentially valuable starch properties without integration of foreign DNA into the genome.     

Photosynthetic genes in viral populations with a large genomic size range from Norwegian coastal waters

This study reports the diversity of uncultured environmental viruses harbouring photosynthetic genes (psbA and psbD) in samples from cold seawater (latitude above 60 degrees ). The viral community in coastal Norwegian waters was separated according to genome size using pulse field gel electrophoresis. Viral populations within a wide genome size range (31-380 kb) were investigated for the presence of the psbA and psbD genes using PCR, combined with cloning and sequencing. The results show the presence of photosynthetic genes in viral populations from all size ranges. Thus, valuable information could be obtained about the size class to which viral particles that encode photosynthesis genes belong. The wide genomic size range detected implies that a different cyanophage profile has been observed than has been reported previously. Thus, the method of phage gene detection applied here may represent a truer picture of phage diversity in general or that there is a larger range of size profile for viruses with psbA and psbD in higher latitudes than for the better-studied lower latitudes. Alternatively, a picture of diversity based on a different set of biases than that from either isolation-based research or from conventional metagenomic approaches may be observed.     

Analysis of host range phenotypes of primate hepadnaviruses by in vitro infections of hepatitis D virus pseudotypes

Hepatitis B virus (HBV) and woolly monkey hepatitis B virus (WMHBV) have natural host ranges that are limited to closely related species. The barrier for infection of primates seems to be at the adsorption and/or entry steps of the viral replication cycle, since a human hepatoma cell line is permissive for HBV and WMHBV replication following transfection of cloned DNA. We hypothesized that the HBV and WMHBV envelope proteins contain the principal viral determinants of host range. As previously shown by using the hepatitis D virus (HDV) system, recombinant HBV-HDV particles were infectious in chimpanzee as well as human hepatocytes. We extended the HDV system to include HDV particles pseudotyped with the WMHBV envelope. In agreement with the natural host ranges of HBV and WMHBV, in vitro infections demonstrated that HBV-HDV and WM-HDV particles preferentially infected human and spider monkey cells, respectively. Previous studies have implicated the pre-S1 region of the large (L) envelope protein in receptor binding and host range; therefore, recombinant HDV particles were pseudotyped with the hepadnaviral envelopes containing chimeric L proteins with the first 40 amino acids from the pre-S1 domain exchanged between HBV and WMHBV. Surprisingly, addition of the human amino terminus to the WMHBV L protein increased infectivity on spider monkey hepatocytes but did not increase infectivity for human hepatocytes. Based upon these data, we discuss the possibility that the L protein may be comprised of two domains that affect infectivity and that sequences downstream of residue 40 may influence host range and receptor binding or entry.