DIXDC1 contributes to psychiatric susceptibility by regulating dendritic spine and glutamatergic synapse density via GSK3 and Wnt/ß-catenin signaling.

Mice lacking DIX domain containing-1 (DIXDC1), an intracellular Wnt/ß-catenin signal pathway protein, have abnormal measures of anxiety, depression and social behavior. Pyramidal neurons in these animals' brains have reduced dendritic spines and glutamatergic synapses. Treatment with lithium or a glycogen synthase kinase-3 (GSK3) inhibitor corrects behavioral and neurodevelopmental phenotypes in these animals. Analysis of DIXDC1 in over 9000 cases of autism, bipolar disorder and schizophrenia reveals higher rates of rare inherited sequence-disrupting single-nucleotide variants (SNVs) in these individuals compared with psychiatrically unaffected controls. Many of these SNVs alter Wnt/ß-catenin signaling activity of the neurally predominant DIXDC1 isoform; a subset that hyperactivate this pathway cause dominant neurodevelopmental effects. We propose that rare missense SNVs in DIXDC1 contribute to psychiatric pathogenesis by reducing spine and glutamatergic synapse density downstream of GSK3 in the Wnt/ß-catenin pathway.

Modeling non-syndromic autism and the impact of TRPC6 disruption in human neurons.

An increasing number of genetic variants have been implicated in autism spectrum disorders (ASDs), and the functional study of such variants will be critical for the elucidation of autism pathophysiology. Here, we report a de novo balanced translocation disruption of TRPC6, a cation channel, in a non-syndromic autistic individual. Using multiple models, such as dental pulp cells, induced pluripotent stem cell (iPSC)-derived neuronal cells and mouse models, we demonstrate that TRPC6 reduction or haploinsufficiency leads to altered neuronal development, morphology and function. The observed neuronal phenotypes could then be rescued by TRPC6 complementation and by treatment with insulin-like growth factor-1 or hyperforin, a TRPC6-specific agonist, suggesting that ASD individuals with alterations in this pathway may benefit from these drugs. We also demonstrate that methyl CpG binding protein-2 (MeCP2) levels affect TRPC6 expression. Mutations in MeCP2 cause Rett syndrome, revealing common pathways among ASDs. Genetic sequencing of TRPC6 in 1041 ASD individuals and 2872 controls revealed significantly more nonsynonymous mutations in the ASD population, and identified loss-of-function mutations with incomplete penetrance in two patients. Taken together, these findings suggest that TRPC6 is a novel predisposing gene for ASD that may act in a multiple-hit model. This is the first study to use iPSC-derived human neurons to model non-syndromic ASD and illustrate the potential of modeling genetically complex sporadic diseases using such cells.

Using large clinical data sets to infer pathogenicity for rare copy number variants in autism cohorts.

Copy number variants (CNVs) have a major role in the etiology of autism spectrum disorders (ASD), and several of these have reached statistical significance in case-control analyses. Nevertheless, current ASD cohorts are not large enough to detect very rare CNVs that may be causative or contributory (that is, risk alleles). Here, we use a tiered approach, in which clinically significant CNVs are first identified in large clinical cohorts of neurodevelopmental disorders (including but not specific to ASD), after which these CNVs are then systematically identified within well-characterized ASD cohorts. We focused our initial analysis on 48 recurrent CNVs (segmental duplication-mediated 'hotspots') from 24 loci in 31 516 published clinical cases with neurodevelopmental disorders and 13 696 published controls, which yielded a total of 19 deletion CNVs and 11 duplication CNVs that reached statistical significance. We then investigated the overlap of these 30 CNVs in a combined sample of 3955 well-characterized ASD cases from three published studies. We identified 73 deleterious recurrent CNVs, including 36 deletions from 11 loci and 37 duplications from seven loci, for a frequency of 1 in 54; had we considered the ASD cohorts alone, only 58 CNVs from eight loci (24 deletions from three loci and 34 duplications from five loci) would have reached statistical significance. In conclusion, until there are sufficiently large ASD research cohorts with enough power to detect very rare causative or contributory CNVs, data from larger clinical cohorts can be used to infer the likely clinical significance of CNVs in ASD.

Whole-exome sequencing in obsessive-compulsive disorder identifies rare mutations in immunological and neurodevelopmental pathways.

Studies of rare genetic variation have identified molecular pathways conferring risk for developmental neuropsychiatric disorders. To date, no published whole-exome sequencing studies have been reported in obsessive-compulsive disorder (OCD). We sequenced all the genome coding regions in 20 sporadic OCD cases and their unaffected parents to identify rare de novo (DN) single-nucleotide variants (SNVs). The primary aim of this pilot study was to determine whether DN variation contributes to OCD risk. To this aim, we evaluated whether there is an elevated rate of DN mutations in OCD, which would justify this approach toward gene discovery in larger studies of the disorder. Furthermore, to explore functional molecular correlations among genes with nonsynonymous DN SNVs in OCD probands, a protein-protein interaction (PPI) network was generated based on databases of direct molecular interactions. We applied Degree-Aware Disease Gene Prioritization (DADA) to rank the PPI network genes based on their relatedness to a set of OCD candidate genes from two OCD genome-wide association studies (Stewart et al., 2013; Mattheisen et al., 2014). In addition, we performed a pathway analysis with genes from the PPI network. The rate of DN SNVs in OCD was 2.51 × 10(-8) per base per generation, significantly higher than a previous estimated rate in unaffected subjects using the same sequencing platform and analytic pipeline. Several genes harboring DN SNVs in OCD were highly interconnected in the PPI network and ranked high in the DADA analysis. Nearly all the DN SNVs in this study are in genes expressed in the human brain, and a pathway analysis revealed enrichment in immunological and central nervous system functioning and development. The results of this pilot study indicate that further investigation of DN variation in larger OCD cohorts is warranted to identify specific risk genes and to confirm our preliminary finding with regard to PPI network enrichment for particular biological pathways and functions.

Apolipophorin-III and adipokinetic hormone in lipid metabolism of larval Manduca sexta.

The hemolymph lipid levels were measured and the density of lipophorin was determined during late larval development in Manduca sexta. During the feeding phase of the 4th and 5th instar larvae the lipid level in hemolymph remained largely unchanged at less than 2 mg/ml. During the molt from 4th to 5th instar, the hemolymph lipid level increased, but decreased after feeding restarted in the 5th instar. In wandering larvae and prepupae the hemolymph lipid level increased from about 2 to nearly 10 mg/ml. The density of lipophorin from feeding larvae was found to be 1.148 g/ml with minor amounts of lipophorin having a lower density of about 1.128 g/ml and sometimes a small amount with a density of 1.174 g/ml. In molting larvae, however, the density was clearly lower, 1.116 g/ml. In wandering larvae of all ages, two predominant forms of lipophorin were observed; the density of these forms was 1.132 g/ml and 1.177 g/ml. Rarely, one or three different forms of lipophorin were observed. While the lipophorin of feeding larvae contains only apoLp-I and II (and lipids), the lipophorin of molting larvae contains in addition apoLp-III. ApoLp-III is seldom present in lipophorin from wandering larvae. According to our current models, lipophorin can take up only a certain amount of diacylglycerol before it needs apoLp-III for surface stabilization. Injection of 1 pmol of M. sexta AKH into feeding larvae increased the hemolymph lipid level, decreased the density of lipophorin to 1.125 g/ml and resulted in the association of apoLp-III with lipophorin. Cardiacectomy did not prevent feeding larvae from developing to wandering larvae.(ABSTRACT TRUNCATED AT 250 WORDS)

Appositions between cocaine and amphetamine-related transcript- and gonadotropin releasing hormone-immunoreactive neurons in the hypothalamus of the Siberian hamster.

Recent in vitro studies have provided evidence that cocaine and amphetamine-related transcript (CART) pathways in the hypothalamus mediate the effects of leptin upon gonadotropin releasing hormone (GnRH) secretion. The aim of the current study was to use dual label immunofluorescence to investigate the anatomical basis of such a pathway. CART-ir processes were found extensively in regions where GnRH cell bodies where located. Analysis using confocal microscopy showed that the majority of GnRH neurons (62%) had close appositions from CART-ir processes. The proportion of GnRH-ir perikarya with CART-ir appositions was significantly higher (P<0.05) in neurons located in the diagonal band of Broca (70%) compared to those more caudally located in the preoptic area (53%). This anatomical evidence for close appositions between CART-ir processes and GnRH cell bodies supports the hypothesis that one mechanism by which leptin causes its effect on the GnRH pulse generator is indirectly via CART neurons, thus allowing information about nutritional status and body fat stores to be conveyed to the reproductive system.

Single-transverse-spin asymmetries of identified charged hadrons in polarized pp collisions at sqrt[s]=62.4 GeV.

The first measurements of xF-dependent single-spin asymmetries of identified charged hadrons, pi+/-, K+/-, and protons, from transversely polarized proton-proton collisions at 62.4 GeV at RHIC are presented. Large asymmetries are seen in the pion and kaon channels. The asymmetries in inclusive pi+ production, AN(pi+), increase with xF from 0 to approximately 0.25 and AN(pi-) decrease from 0 to approximately -0.4. Observed asymmetries for K- unexpectedly show positive values similar to those for K+, increasing with xF, whereas proton asymmetries are consistent with zero over the measured kinematic range. Comparisons of the data with predictions of QCD-based models are presented.

Production of mesons and baryons at high rapidity and high p(T) in proton-proton collisions at square root[s] = 200 GeV.

We present particle spectra for charged hadrons pi(+/-), K(+/-), p, and p[over] from pp collisions at square root[s] = 200 GeV measured for the first time at forward rapidities (2.95 and 3.3). The kinematics of these measurements are skewed in a way that probes the small momentum fraction in one of the protons and large fractions in the other. Large proton to pion ratios are observed at values of transverse momentum that extend up to 4 GeV/c, where protons have momenta up to 35 GeV. Next-to-leading order perturbative QCD calculations describe the production of pions and kaons well at these rapidities, but fail to account for the large proton yields and small p[over]/p ratios.

Centrality dependence of charged-particle pseudorapidity distributions from d + Au collisions at sqrt[sNN] = 200 GeV.

Charged-particle pseudorapidity densities are presented for the d + Au reaction at sqrt[s(NN)] = 200 GeV with -4.2 < or = eta < or = 4.2. The results, from the BRAHMS experiment at BNL Relativistic Heavy-Ion Collider, are shown for minimum-bias events and 0%-30%, 30%-60%, and 60%-80% centrality classes. Models incorporating both soft physics and hard, perturbative QCD-based scattering physics agree well with the experimental results. The data do not support predictions based on strong-coupling, semiclassical QCD. In the deuteron-fragmentation region the central 200 GeV data show behavior similar to full-overlap d+Au results at sqrt[s(NN)] = 19.4 GeV.

Charged meson rapidity distributions in central Au+Au collisions at square root(sNN) = 200 GeV.

We have measured rapidity densities dN/dy of pi+/- and K+/- over a broad rapidity range (-0.1 < y < 3.5) for central Au + Au collisions at square root(sNN) = 200 GeV. These data have significant implications for the chemistry and dynamics of the dense system that is initially created in the collisions. The full phase-space yields are 1660 +/- 15 +/- 133 (pi+), 1683 +/- 16 +/- 135 (pi-), 286 +/- 5 +/- 23 (K+), and 242 +/- 4 +/- 19 (K-). The systematics of the strange to nonstrange meson ratios are found to track the variation of the baryochemical potential with rapidity and energy. Landau-Carruthers hydrodynamics is found to describe the bulk transport of the pions in the longitudinal direction.

Hereditary hyperferritinaemia-cataract syndrome and differential diagnosis of hereditary haemochromatosis.

Hereditary hyperferritinaemia-cataract syndrome (HHCS) is a rare differential diagnosis of hereditary haemochromatosis. It should be suspected in patients with raised ferritin levels, but no evidence of iron overload, and in the absence of mutations in the HFE gene. Awareness of this condition prevents unnecessary liver biopsies and allows accurate genetic counselling since HHCS is an autosomal dominant disorder. The danger of treating these patients by phlebotomy in the same manner as those with hereditary haemochromatosis is highlighted.

Serum bactericidal testing with the Autobac system.

Current methodology for the serum bactericidal test requires a minimum of 48 h. A procedure was devised for performing this test with the Autobac system (General Diagnostics, Div. Organon Inc., Raleigh, N.C.) in a shortened time span. All titers obtained with the Autobac were compared against results obtained with a standardized tube dilution procedure. The Autobac low-thymidine eugonic broth performed comparably to the tube dilution diluent, a 1:1 ratio of pooled human serum and cation-supplemented Mueller-Hinton broth (99.2% correlation between bactericidal endpoints). Over 300 tests were conducted by using stock reference bacterial strains, clinical isolates, pooled human serum seeded with antimicrobial agents, and serum from patients on antimicrobial therapy. With the Autobac procedure, serum inhibitory titers can be reported in 3 to 4 h (93.4% correlation with the tube dilution procedure). Serum bactericidal titers can be obtained in 24 h without the necessity of subculturing (95.6% correlation). With the exception of staphylococci tested against penicillin, serum bactericidal titers can be obtained in 3 to 4 h (88.4% correlation). The Autobac procedure can provide the clinical laboratory with a rapid, reliable method for performing the serum bactericidal test.

Empirical investigation of extreme single-particle behavior of nuclear quadrupole moments in highly collective A approximately 150 superdeformed bands.

The intrinsic quadrupole moment Q(0) of superdeformed rotational bands in A approximately 150 nuclei depends on the associated single-particle configuration. We have derived an empirical formula based on the additivity of effective quadrupole moments of single-particle orbitals that describes existing measurements from (142)Sm to (152)Dy. To further test the formula, the predicted Q(0) moments for two superdeformed bands in (146)Gd of 14.05 eb were confronted with a new measurement yielding 13.9+/-0.4 eb and 13.9+/-0.3 eb, respectively. This excellent agreement provides empirical evidence of extreme single-particle behavior in highly deformed, collective systems.

Pseudorapidity distributions of charged particles from Au + Au collisions at the maximum RHIC energy, square root[s(NN)] = 200 GeV.

We present charged-particle multiplicities as a function of pseudorapidity and collision centrality for the 197Au+197Au reaction at square root[s(NN)] = 200 GeV. For the 5% most central events we obtain dN(ch)/deta/(eta = 0) = 625+/-55 and N(ch)/(-4.7< or =eta < or =4.7) = 4630 +/- 370, i.e., 14% and 21% increases, respectively, relative to square root[s(NN)] = 130 GeV collisions. Charged-particle production per pair of participant nucleons is found to increase from peripheral to central collisions around midrapidity. These results constrain current models of particle production at the highest RHIC energy.

Nuclear stopping in Au+Au collisions at square root of S(NN)=200 GeV.

Transverse momentum spectra and rapidity densities, dN/dy, of protons, antiprotons, and net protons (p-p) from central (0%-5%) Au+Au collisions at square root of S(NN)=200 GeV were measured with the BRAHMS experiment within the rapidity range 0

Rapidity dependence of charged antihadron to hadron ratios in Au+Au collisions at sqrt[s(NN)]=200 GeV.

We present ratios of the numbers of charged antihadrons to hadrons (pions, kaons, and protons) in Au+Au collisions at sqrt[s(NN)]=200 GeV as a function of rapidity in the range y=0-3. While the ratios at midrapidity are approaching unity, the K(-)/K(+) and p;/p ratios decrease significantly at forward rapidities. An interpretation of the results within the statistical model indicates a reduction of the baryon chemical potential from mu(B) approximately 130 MeV at y=3 to mu(B) approximately 25 MeV at y=0.

Transverse-momentum spectra in Au+Au and d+Au collisions at sqrt[s(NN)]=200 GeV and the pseudorapidity dependence of high-p(T) suppression.

We present spectra of charged hadrons from Au+Au and d+Au collisions at sqrt[s(NN)]=200 GeV measured with the BRAHMS experiment at RHIC. The spectra for different collision centralities are compared to spectra from p+(-)p collisions at the same energy scaled by the number of binary collisions. The resulting ratios (nuclear modification factors) for central Au+Au collisions at eta=0 and eta=2.2 evidence a strong suppression in the high p(T) region (>2 GeV/c). In contrast, the d+Au nuclear modification factor (at eta=0) exhibits an enhancement of the high p(T) yields. These measurements indicate a high energy loss of the high p(T) particles in the medium created in the central Au+Au collisions. The lack of suppression in d+Au collisions makes it unlikely that initial state effects can explain the suppression in the central Au+Au collisions.

Evolution of the nuclear modification factors with rapidity and centrality in d + Au collisions at (sqrt)[N(S)N]=200 GeV.

We report on a study of the transverse momentum dependence of nuclear modification factors R(dAu) for charged hadrons produced in deuteron + gold collisions at sqrt[s(NN)]=200 GeV, as a function of collision centrality and of the pseudorapidity (eta=0, 1, 2.2, 3.2) of the produced hadrons. We find a significant and systematic decrease of R(dAu) with increasing rapidity. The midrapidity enhancement and the forward rapidity suppression are more pronounced in central collisions relative to peripheral collisions. These results are relevant to the study of the possible onset of gluon saturation at energies reached at BNL RHIC.