Stem cell-derived neurons from autistic individuals with SHANK3 mutation show morphogenetic abnormalities during early development.

Shank3 is a structural protein found predominantly at the postsynaptic density. Mutations in the SHANK3 gene have been associated with risk for autism spectrum disorder (ASD). We generated induced pluripotent stem cells (iPSCs) from control individuals and from human donors with ASD carrying microdeletions of SHANK3. In addition, we used Zinc finger nucleases to generate isogenic SHANK3 knockout human embryonic stem (ES) cell lines. We differentiated pluripotent cells into either cortical or olfactory placodal neurons. We show that patient-derived placodal neurons make fewer synapses than control cells. Moreover, patient-derived cells display a developmental phenotype: young postmitotic neurons have smaller cell bodies, more extensively branched neurites, and reduced motility compared with controls. These phenotypes were mimicked by SHANK3-edited ES cells and rescued by transduction with a Shank3 expression construct. This developmental phenotype is not observed in the same iPSC lines differentiated into cortical neurons. Therefore, we suggest that SHANK3 has a critical role in neuronal morphogenesis in placodal neurons and that early defects are associated with ASD-associated mutations.

Mechanisms for pressure-induced crystal-crystal transition, amorphization, and devitrification of SnI4.

The pressure-induced amorphization and subsequent recrystallization of SnI4 have been investigated using first principles molecular dynamics calculations together with high-pressure (119)Sn nuclear resonant inelastic x-ray scattering measurements. Above ∼8 GPa, we observe a transformation from an ambient crystalline phase to an intermediate crystal structure and a subsequent recrystallization into a cubic phase at ∼64 GPa. The crystalline-to-amorphous transition was identified on the basis of elastic compatibility criteria. The measured tin vibrational density of states shows large amplitude librations of SnI4 under ambient conditions. Although high pressure structures of SnI4 were thought to be determined by random packing of equal-sized spheres, we detected electron charge transfer in each phase. This charge transfer results in a crystal structure packing determined by larger than expected iodine atoms.

Determination of iron sites and the amount of amorphization in radiation-damaged titanite (CaSiTiO5).

Iron is a ubiquitous impurity in metamict (radiation-damaged and partially amorphized) materials such as titanite (CaSiTiO(5)). Using (57)Fe Mössbauer spectroscopy we find that iron in metamict titanite is partitioned between amorphous and crystalline regions based on valence. Trivalent iron exists in the crystalline titanite matrix whereas divalent iron exists almost exclusively in radiation-amorphized regions. We find that the relative abundances of the oxidation states correlate with the volume fraction of amorphous and crystalline regions. Our data also show that oxidation of iron proceeds along with the recrystallization of the amorphized regions. Recrystallization is confirmed to occur over the range 700 °C < T < 925 °C, and no further structural changes are observed at higher temperatures. It is surprising that our Mössbauer measurements show divalent iron to be surrounded by titanite with a high degree of short-range structural order in the amorphized regions. This observation is fundamentally different from other metamict materials such as zircon (ZrSiO(4)), where amorphized regions show no short-range order.

Dog obesity: veterinary practices' and owners' opinions on cause and management.

Obesity in dogs is attributed to several factors, including genetic pre-disposition, reproductive management and dietary/exercise (human) management. A quantitative analysis of questionnaire responses from dog owners and veterinary practices in Victoria, Australia was used. A total of 219 dog owner questionnaire and 153 veterinary practice questionnaires were returned. Veterinary practices estimated the prevalence of dog obesity within their practice as 30% on average, ranging from <10 to 100%. Veterinary practices felt that 3% of cases could be attributed to dog specific factors and 97% to human specific factors such as diet, exercise and owner attitudes. Management strategies included reducing food intake, reduced treat feeding and changing diet before recommending more intensive options such as obesity clinics. Of the surveyed veterinary practices, 43% ran obesity clinics and 79% of those believed they were a valuable management tool. Of veterinary practices that did not run obesity clinics, only 46% believed them to be a valuable management tool. Dog owners preferred to try to reduce dog weight through diet manipulation, increasing exercise and elimination of treats prior to consulting the veterinary practice. There is a clear difference in expectations with veterinary practices recommending reduction in dietary intake compared to owners expecting to have to modify the diet itself, which may have a cost and hence psychological implication to the owner.

Dog obesity: owner attitudes and behaviour.

Animal (dog) factors that contribute to obesity are classified into three broad categories: genetic pre-disposition, reproductive management and dietary/exercise (human) management. This paper examined the latter-dietary/exercise (human) management. A quantitative analysis of questionnaire responses from dog owners and veterinarians was used to determine the routine care and obesity management strategies for dogs. A total of 550 questionnaires were distributed to dog owners in Victoria, Australia. Owners were asked to score the body condition of their animal by comparison with photographic images of animals with condition score ranging from 2 to 5. The management routines of 219 dog owners were received, including data on 302 dogs. There were 168 households with normal weight animals (BCS 2 and 3) and 51 with obese animals (BCS 4 and 5). The mean number of people per household (normally involved with caring for the animal(s)) with normal weight dogs was significantly lower than that of households with dogs categorised as overweight or obese (Kruskal-Wallis, Chi; chi(2)=6.28; 2.2 (s=0.79) vs. 2.5 (s=1.66); d.f.=2, P<0.05). Dog owners identified a preference for main meal feeding of 'twice a day' (60%), followed by 'once daily' (33%), 'greater than or equal to three times daily' (2%), and 'always feed available' (5%). There was a significant difference (Chi; chi(2)=6.30; d.f.=1; P<0.05) in the frequency of main meal feeding between households. Normal weight animals had food divided into two portions, whereas obese animals or animals from mixed households were more often fed their meal in either one or three-plus portions. Almost all owners fed treats (99%) in the daily feed. Households with normal weight dogs gave treats significantly less frequently than households with obese or mixed weight dogs (Chi; chi(2)=31.81; d.f.=6; P<0.001). The frequency of exercise differed between households (Chi; chi(2)=9.9; d.f.=1; P<0.01), with normal weight dogs being exercised daily compared to weekly for overweight or mixed weight dogs. Owners who indicated that their dog was confined to a yard as its exercise regime rather than walked were also significantly more likely to be an obese or mixed weight household (Chi; chi(2)=18.4; d.f.=1; P<0.001). In conclusion, there is clear evidence that obesity in dogs is affected by the interrelationships between food management, exercise and social factors.

Differential growth and development of pigs as assessed by X-ray computed tomography.

Data from 54 hybrid (mainly Large White x Landrace) pigs (18 boars, 18 gilts, and 18 barrows) were used to quantify and mathematically describe the differential growth and development of body components of live pigs. The pigs were 32.4 +/- 3.2 kg of BW and 70 +/- 1 d of age (mean +/- SD) at the beginning of the study, were individually penned and fed ad libitum, and were weighed weekly. Computed tomography (CT) imaging was used to determine the weights of lean, fat, bone, and skin tissue in the live pig at 30, 60, 90, 120, and 150 kg of BW. For each target BW, the sum of all the weights of the body components, as assessed by CT, was referred to as CT BW. Linear and nonlinear models were developed to evaluate the patterns of growth and development of each body component relative to CT BW. The correlation between the actual BW and CT BW was close to unity (r = 0.99), indicating that CT scanning could accurately predict the BW of pigs. Across sex and castrate status, percentage of fat (fat weight/CT BW) in the pig was least (11.2%) at the 30-kg target BW and continued to increase to 22.6% by the 150-kg target BW. Percentage of lean, however, was greatest (67.2%) at the 30-kg target BW and continued to decrease to 53.4% by the 150-kg target BW. The sex or castrate status x target BW interaction was significant (P < 0.05) for all the body components, indicating that the developmental patterns were different among sex or castrate status. Barrows were fatter relative to gilts, which in turn were fatter than boars. For lean, the observed pattern for sex or castrate status differences was opposite that for fat. To predict responses to management strategies on growth and development in pigs, accurate mathematical models are required, and the results of this study indicate that the nonlinear (e.g., augmented allometric and generalized nonlinear) functions provided better descriptions of the growth and development of most body components of the live pig than did the simpler (e.g., linear and allometric) models.

Origin of the Verwey transition in magnetite.

Comprehensive x-ray powder diffraction studies were carried out in magnetite in the 80-150 K and 0-12 GPa ranges with a membrane-driven diamond anvil cell and helium as a pressure medium. Careful data analyses have shown that a reversible, cubic to a distorted-cubic, structural transition takes place with increasing pressure, within the (P,T) regime below the Verwey temperature TV(P). The experimental documentation that TV(P)=Tdist(P) implies that the pressure-temperature-driven metal-insulator Verwey transition is caused by a gap opening in the electronic band structure due to the crystal-structural transformation to a lower-symmetry phase. The distorted-cubic insulating phase comprises a relatively small pressure-temperature range of the stability field of the cubic metallic phase that extends to 25 GPa.

The incorporation of whole grain into pelleted broiler chicken diets. II. Gastrointestinal and digesta characteristics.

1. Pelleted diets, incorporating whole or ground wheat or barley in the pellets, were fed to broiler chickens and the performance, gastrointestinal development and digesta characteristics of those chickens recorded. 2. Body weight was similar with whole grain incorporation or enzyme application. Food conversion efficiency was improved by addition of enzyme to a full wheat diet. 3. Whole grain inclusion in pellets reduced proventriculus proportional mass and increased gizzard proportional mass with no apparent effects of exogenous feed enzyme addition. 4. Relative ileal mass was reduced by enzyme inclusion in a wheat diet and by inclusion of whole barley in the pellets. 5. Digesta viscosity was increased by whole wheat inclusion but reduced by enzyme inclusion irrespective of grain processing. Inclusion of 200 g/kg of barley did not alter viscosity of digesta beyond the duodenum. 6. Fresh excreta pH was higher with both cereals in the grower phase when whole grain was fed. On the barley diet, this was affected by enzyme addition, which created higher pH from a point earlier in the grower phase than when no enzyme was employed. 7. Evidence of a complex interaction between higher viscosity and pH being involved in differences in ileal relative mass was found through significant relationships being produced by enzyme use on the wheat diet alone.

The influence of whole grain inclusion in pelleted broiler diets on proventricular dilatation and ascites mortality.

1. Pelleted diets, incorporating whole or ground wheat or barley in the pellets, were fed to broiler chickens and proventricular dilatation, digesta characteristics and mortality were recorded. 2. The incidence of proventricular dilatation was reduced when whole grain was incorporated into pelleted diets at 200 g/kg. 3. Enzyme application did not influence proventricular dilatation. 4. Digesta contents of intestinal sections were greater and pH lower in anterior intestinal sections when birds had dilated proventriculi. 5. Consideration of overall ascites mortality suggests that the incidence of the condition may be curtailed by whole grain inclusion in pelleted diets. 6. Production efficiency may be enhanced with whole grain inclusion in pellets when feed processing costs and flock health responses are considered.

Measured weights in PEI adults reveal higher than expected obesity rates.

BACKGROUND: The increasing prevalence of obesity and overweight in Canada is a significant health concern. Unfortunately, we know very little about the actual weight status and associated health risks in our population since most surveys use only self-reported body weights and heights and typically do not include a measure of body fat distribution. This paper summarizes the findings of the Prince Edward Island Nutrition Survey. METHODS: A random sample of 1,995 adults aged 18-74 were interviewed in their homes and weights, heights and waist circumference measurements were obtained. Relative health risks, population proportions and their corresponding 95% confidence intervals were calculated. RESULTS: Overall, almost one third of PEI adults are obese (BMI > or = 30). This is almost double that reported in the 1995 National Population Health Survey using self-reported heights and weights. More women were classified as being very severely obese (Class III) than men, but for both men and women there appears to be a trend of increased mild obesity with age. Based on BMI and waist circumference, over one third of the population is considered to be at high to extremely high risk for health problems. CONCLUSION: Self-reported height and weight data appear to result in significant underestimation of the problem of obesity. Given the serious health consequences associated with this condition, it is critical that measured heights and weights be collected in future population-based surveys to ensure that public health interventions are based on accurate prevalence data.

Pressure induced self-oxidation of Fe(OH)2.

Mössbauer spectroscopy, x-ray diffraction, and electrical resistance [R(P,T)] studies in Fe(OH)(2) to 40 GPa revealed an unforeseen process by which a gradual Fe2+ oxidation takes place, starting at approximately 8 GPa reaching 70% Fe3+ abundance at 40 GPa. The nonreversible process Fe2+-->Fe3++e(-) occurs with no structural transition. The "ejected" electrons form a deep band within the high-pressure electronic manifold becoming weakly localized at P>50 GPa. This process is attributed to an effective ionization potential created by the pressure induced orientationally deformed (OH) dipoles and the unusual small binding energy of the valence electron in Fe2+(OH)(2).

A review of protein-small molecule docking methods.

The binding of small molecule ligands to large protein targets is central to numerous biological processes. The accurate prediction of the binding modes between the ligand and protein, (the docking problem) is of fundamental importance in modern structure-based drug design. An overview of current docking techniques is presented with a description of applications including single docking experiments and the virtual screening of databases.

The incorporation of whole grain into pelleted broiler chicken diets: production and physiological responses.

1. Pelleted diets, incorporating whole or ground triticale or wheat in the pellets, were fed to broiler chickens and the performance, gastro-intestinal development and health of those chickens recorded. 2. The use of whole triticale in the pelleted food produced similar bodyweight responses to when ground triticale was incorporated in the food. Food conversion efficiency was enhanced when whole triticale was used compared to ground triticale and was similar to when an exogenous enzyme was added to the ground triticale diet. The incorporation of whole wheat into the pelleted food produced similar production responses to the use of ground wheat. 3. The gastro-intestinal development of the birds was greater when they were fed the whole grain diet as gizzard weights were increased by the incorporation of whole grain into the diet. Gastro-intestinal development was thought to be related to the presence of large fibre particles in the food. 4. Proventricular dilatation and mortality due to ascites were reduced by feeding pelleted diets containing whole grain, which was possibly related to the early development of the gastro-intestinal tract relative to total body development. 5. It is suggested that the current use of exogenous enzyme additions to broiler diets may be reduced by incorporating whole grain into pelleted diets for broiler chickens.

TANK2, a new TRF1-associated poly(ADP-ribose) polymerase, causes rapid induction of cell death upon overexpression.

Tankyrase (TANK1) is a human telomere-associated poly(ADP-ribose) polymerase (PARP) that binds the telomere-binding protein TRF1 and increases telomere length when overexpressed. Here we report characterization of a second human tankyrase, tankyrase 2 (TANK2), which can also interact with TRF1 but has properties distinct from those of TANK1. TANK2 is encoded by a 66-kilobase pair gene (TNKS2) containing 28 exons, which express a 6.7-kilobase pair mRNA and a 1166-amino acid protein. The protein shares 85% amino acid identity with TANK1 in the ankyrin repeat, sterile alpha-motif, and PARP catalytic domains but has a unique N-terminal domain, which is conserved in the murine TNKS2 gene. TANK2 interacted with TRF1 in yeast and in vitro and localized predominantly to a perinuclear region, similar to the properties of TANK1. In contrast to TANK1, however, TANK2 caused rapid cell death when highly overexpressed. TANK2-induced death featured loss of mitochondrial membrane potential, but not PARP1 cleavage, suggesting that TANK2 kills cells by necrosis. The cell death was prevented by the PARP inhibitor 3-aminobenzamide. In vivo, TANK2 may differ from TANK1 in its intrinsic or regulated PARP activity or its substrate specificity.

Structural requirements of Tom40 for assembly into preexisting TOM complexes of mitochondria.

Tom40 is the major subunit of the translocase of the outer mitochondrial membrane (the TOM complex). To study the assembly pathway of Tom40, we have followed the integration of the protein into the TOM complex in vitro and in vivo using wild-type and altered versions of the Neurospora crassa Tom40 protein. Upon import into isolated mitochondria, Tom40 precursor proteins lacking the first 20 or the first 40 amino acid residues were assembled as the wild-type protein. In contrast, a Tom40 precursor lacking residues 41 to 60, which contains a highly conserved region of the protein, was arrested at an intermediate stage of assembly. We constructed mutant versions of Tom40 affecting this region and transformed the genes into a sheltered heterokaryon containing a tom40 null nucleus. Homokaryotic strains expressing the mutant Tom40 proteins had growth rate defects and were deficient in their ability to form conidia. Analysis of the TOM complex in these strains by blue native gel electrophoresis revealed alterations in electrophoretic mobility and a tendency to lose Tom40 subunits from the complex. Thus, both in vitro and in vivo studies implicate residues 41 to 60 as containing a sequence required for proper assembly/stability of Tom40 into the TOM complex. Finally, we found that TOM complexes in the mitochondrial outer membrane were capable of exchanging subunits in vitro. A model is proposed for the integration of Tom40 subunits into the TOM complex.

Proneness to eating disorders: weightlifters compared to exercisers.

Previous findings were confirmed suggesting that weightlifters may be more prone to eating disorders than are casual exercisers. 25 weightlifters' mean eating disorder score was significantly higher than that of 25 casual exercisers when contrasted for practical difference.

Influence of the method of calcium provision on Marek's disease losses in compound and choice fed layers.

1. Two strains of layers were fed compound or choice forms of a standard diet and given calcium as either ground limestone or coarse limestone grit daily or every 2nd day. Mortality caused by Marek's disease was monitored during the rearing and laying phases. 2. A dramatic difference in Marek's disease mortality was found between the strains. Heavy Marek's disease mortality in the imported strain A in the rearing phase was not markedly affected by dietary or calcium treatment. 3. Method of feeding and calcium provision interacted to influence significantly the risk of death due to Marek's disease in the laying phase in layer strain A. 4. Choice feeding may adversely influence Marek's disease losses in layers.

U-richness is a defining feature of plant introns and may function as an intron recognition signal in maize.

Using a large set of plant gene sequences we compared individual introns to their flanking exons. Both Zea mays and Arabidopsis thaliana introns are U-rich but display no apparent bias for A. We identified fifteen 11-mer U-rich motifs as frequent elements of maize introns, and these are virtually absent from exons. By mutagenesis, we show that the single U-rich motif in the Bronze2 intron of maize plays a key role in intron processing in vivo.

Reconstitution of human telomerase with the template RNA component hTR and the catalytic protein subunit hTRT.

The maintenance of chromosome termini, or telomeres, requires the action of the enzyme telomerase, as conventional DNA polymerases cannot fully replicate the ends of linear molecules. Telomerase is expressed and telomere length is maintained in human germ cells and the great majority of primary human tumours. However, telomerase is not detectable in most normal somatic cells; this corresponds to the gradual telomere loss observed with each cell division. It has been proposed that telomere erosion eventually signals entry into senescence or cell crisis and that activation of telomerase is usually required for immortal cell proliferation. In addition to the human telomerase RNA component (hTR; ref. 11), TR1/TLP1 (refs 12, 13), a protein that is homologous to the p80 protein associated with the Tetrahymena enzyme, has been identified in humans. More recently, the human telomerase reverse transcriptase (hTRT; refs 15, 16), which is homologous to the reverse transcriptase (RT)-like proteins associated with the Euplotes aediculatus (Ea_p123), Saccharomyces cerevisiae (Est2p) and Schizosaccharomyces pombe (5pTrt1) telomerases, has been reported to be a telomerase protein subunit. A catalytic function has been demonstrated for Est2p in the RT-like class but not for p80 or its homologues. We now report that in vitro transcription and translation of hTRT when co-synthesized or mixed with hTR reconstitutes telomerase activity that exhibits enzymatic properties like those of the native enzyme. Single amino-acid changes in conserved telomerase-specific and RT motifs reduce or abolish activity, providing direct evidence that hTRT is the catalytic protein component of telomerase. Normal human diploid cells transiently expressing hTRT possessed telomerase activity, demonstrating that hTRT is the limiting component necessary for restoration of telomerase activity in these cells. The ability to reconstitute telomerase permits further analysis of its biochemical and biological roles in cell aging and carcinogenesis.

Kinship support and maternal and adolescent well-being in economically disadvantaged African-American families.

This study tested a conceptual model developed to explain the link between kinship support and the psychological well-being of economically disadvantaged African-American adolescents. The relation of kinship support with maternal and adolescent well-being and mothers' child-rearing practices was assessed in 51 African-American families whose incomes placed them at or below the poverty threshold. Findings revealed that kinship social support to mothers/female guardians was positively associated with adolescent psychological well-being, maternal well-being, and more adequate maternal parenting practices (acceptance, firm control and monitoring of behavior, autonomy granting). Maternal well-being and more adequate maternal parenting practices were positively related to adolescent well-being. Evidence of the mediational role of maternal well-being and parenting practices was revealed. When the effects of maternal well-being and maternal parenting practices were controlled, significant relations between kinship support and adolescent well-being were no longer apparent.