On-chip manipulation of single photons from a diamond defect.

Operating reconfigurable quantum circuits with single photon sources is a key goal of photonic quantum information science and technology. We use an integrated waveguide device containing directional couplers and a reconfigurable thermal phase controller to manipulate single photons emitted from a chromium related color center in diamond. Observation of both a wavelike interference pattern and particlelike sub-Poissionian autocorrelation functions demonstrates coherent manipulation of single photons emitted from the chromium related center and verifies wave particle duality.

Aging in the cerebellum and hippocampus and associated behaviors over the adult life span of CB6F1 mice.

In the present study we examined the effects of normal aging in the hippocampus and cerebellum, as well as behaviors associated with these substrates. A total of 67 CB6F1 hybrid mice were tested at one of five ages (4, 8, 12, 18 or 25 months) on the context pre-exposure facilitation effect (CPFE) modification of fear conditioning, rotorod, Barnes maze, acoustic startle, Morris water maze (MWM) and 500-ms trace eyeblink classical conditioning (EBCC). Behavioral tasks were chosen to increase the ability to detect age-related changes in learning, as trace EBCC is considered a more difficult paradigm (compared to delay EBCC) and the CPFE has been found to be more sensitive to hippocampus insults than standard contextual fear conditioning. To assess the effects of age on the brain, hippocampus volume was calculated and unbiased stereology was used to estimate the number of Purkinje neurons in the cerebellar cortex. A significant, age-related loss of Purkinje neurons was found-beginning at 12 months of age-and hippocampus volume remained stable over the adult life span. Age-related impairment was found, beginning at 12-18 months in the rotorod, and mice with fewer Purkinje neurons showed greater impairment in this task. CB6F1 mice retained auditory acuity across the life span and mice aged 25 months showed significant age-related impairment in the EBCC task; however, deficits were not associated with the loss of Purkinje neurons. Although the CPFE task is considered more sensitive to hippocampus insult, no age-related impairment was found. Spatial memory retention was impaired in the Barnes maze at 25 months, but no significant deficits were seen in the MWM. These results support the finding of differential aging in the hippocampus and cerebellum.

Aging affects L-arginine and its metabolites in memory-associated brain structures at the tissue and synaptoneurosome levels.

L-arginine, one of the most metabolically versatile amino acids, can be metabolized to form a number of bioactive molecules. The present study systematically investigated age-related changes in L-arginine and its metabolites in the hippocampus, parahippocampal region, and prefrontal cortex at the tissue (crude homogenates) and synaptoneurosome (a subcellular preparation enriched for synaptic material) levels. As aging leads to reduced water content in the brain, age-related changes in neurochemical levels in tissue homogenates normalized by wet tissue weight and protein level were compared. There were significant differences in L-arginine, L-citrulline, L-ornithine, agmatine, putrescine, spermidine, spermine, and glutamate, but not GABA, in the CA1, CA2/3, and dentate gyrus sub-regions of the hippocampus and the prefrontal, entorhinal, perirhinal, and postrhinal cortices in 24 (aged) and 4 (young) months old rats in a region-specific manner. The overall pattern of age-related changes in amino acids (L-arginine, L-citrulline, L-ornithine, glutamate, and GABA) was largely similar between homogenates and synaptoneurosomes, whereas the pattern for the amines (agmatine, putrescine, spermidine, and spermine) was quite different. Furthermore, the pattern of age-related changes in neurochemical levels in tissue homogenates normalized by wet tissue weight and protein level was very similar for all 9 neurochemicals measured. These findings suggest that there are differential effects of aging on L-arginine metabolism at the tissue and synaptoneurosome levels and that the way of data normalization (tissue weight vs. protein level) has no or very minor effects on 9 neurochemicals measured.

Stargazin and AMPA receptor membrane expression is increased in the somatosensory cortex of Genetic Absence Epilepsy Rats from Strasbourg.

Absence-like seizures in the Genetic Absence Epilepsy Rats from Strasbourg (GAERS) model are believed to arise in hyperexcitable somatosensory cortical neurons, however the cellular basis of this increased excitability remains unknown. We have previously shown that expression of the Transmembrane AMPA receptor Regulatory Protein (TARP), stargazin, is elevated in the somatosensory cortex of GAERS. TARPs are critical regulators of the trafficking and function of AMPA receptors. Here we examine the developmental expression of stargazin and the impact this may have on AMPA receptor trafficking in the GAERS model. We show that elevated stargazin in GAERS is associated with an increase in AMPA receptor proteins, GluA1 and GluA2 in the somatosensory cortex plasma membrane of adult epileptic GAERS. Elevated stargazin expression is not seen in the epileptic WAG/Rij rat, which is a genetically distinct but phenotypically similar rat model also manifesting absence seizures, indicating that the changes seen in GAERS are unlikely to be a secondary consequence of the seizures. In juvenile (6 week old) GAERS, at the age when seizures are just starting to be expressed, there is elevated stargazin mRNA, but not protein expression for stargazin or the AMPA receptor subunits. In neonatal (7 day old) pre-epileptic GAERS there was no alteration in stargazin mRNA expression in any brain region examined. These data demonstrate that stargazin and AMPA receptor membrane targeting is altered in GAERS, potentially contributing to hyperexcitability in somatosensory cortex, with a developmental time course that would suggest a pathophysiological role in the epilepsy phenotype.

Long-term regulation of N-methyl-D-aspartate receptor subunits and associated synaptic proteins following hippocampal synaptic plasticity.

Synaptic plasticity in the dentate gyrus is dependent on activation of the N-methyl-D-aspartate (NMDA)-subtype of glutamate receptors. In this study, we show that synaptic plasticity in turn regulates NMDA receptors, since subunits of the NMDA receptor complex are bidirectionally and independently regulated in the dentate gyrus following activation of perforant synapses in awake animals. Low-frequency stimulation that produced a mild synaptic depression resulted in a decrease in the NMDA receptor subunits NR1 and NR2B 48 h following stimulation. High-frequency stimulation that produced long-term potentiation resulted in an increase in NR1 and NR2B at the same time point. Further investigations revealed that in contrast to NR2B, NR1 levels increased gradually after long-term potentiation induction, reaching a peak level at 48 h, and were insensitive to the competitive NMDA receptor antagonist 3-3(2-carboxypiperazin-4-yl) propyl-1-phosphate. The increased levels of NR1 and NR2B at 48 h were found associated with synaptic membranes and with increased NMDA receptor-associated proteins, postsynaptic density protein 95, neuronal nitric oxide synthase and Ca(2+)/calmodulin-dependent protein kinase II, alpha subunit. These data suggest that the persistence of long-term potentiation is associated with an increase in the number of NMDA receptor complexes, which may be indicative of an increase in synaptic contact area.

Efficient herpes simplex virus type 1 (HSV-1) capsid formation directed by the varicella-zoster virus scaffolding protein requires the carboxy-terminal sequences from the HSV-1 homologue.

The scaffolding protein and associated protease of the human herpesvirus varicella-zoster virus (VZV), encoded by genes 33.5 and 33 respectively, were synthesized in insect cells using a baculovirus expression system. The expressed 33.5 product formed numerous long, flexible, hollow rods, and in this respect different from the herpes simplex virus type 1 (HSV-1) homologue which forms large aggregates consisting mainly of fibrous material interspersed with scaffold-like particles. Removal of 27 amino acids from the carboxy terminus of the VZV scaffolding protein by the gene 33 protease or expression of the cleaved product did not result in any discernible change in the morphology of the scaffolding protein. Again, this was in marked contrast to the situation in HSV-1 where removal of the 25 carboxy-terminal amino acids from the scaffolding protein by the associated protease or expression of VP22a results in the formation of large numbers of scaffold-like particles. Despite these differences, when cells were multiply infected with baculoviruses expressing the HSV-1 capsid shell proteins and the VZV scaffolding protein complete capsids were observed, suggesting that the VZV protein could act as a scaffold for the assembly of the HSV-1 capsid shell. The efficiency of capsid assembly was increased substantially by exchanging the 23 carboxy-terminal amino acids of the VZV scaffolding protein for the corresponding 22 carboxy-terminal amino acids of the HSV-1 homologue, supporting previous work which showed that this region was critical for the formation of intact capsids.

Pre-mitotic nuclear migration in subsidiary mother cells of Tradescantia occurs in G1 of the cell cycle and requires F-actin.

We have studied pre-mitotic nuclear migration in living subsidiary mother cells (SMCs) of Tradescantia virginiana. Divisions in the four SMCs of each stomatal complex are asymmetrical and are preceded by the migration of nuclei from random locations in the cells to positions adjacent to the central guard mother cells (GMCs). In newly polarised SMCs, nuclei display erratic movements which gradually dampen over time. In older complexes, where nuclear migration occurred earlier in the ontogeny of the leaf, nuclei are stably positioned and change in morphology from spherical to dome-shaped. Labelling with bromodeoxyuridine (BrdU) shows that SMCs polarised in G1 of the cell cycle and remain polarised for a minimum of 22 h before entering mitosis, while the inducing GMCs stay in G1. Centrifugation (1,320 g, 15 min) of epidermal peels displaces the majority of nuclei to the centrifugal end of cells, including nuclei in polarised SMCs. After centrifugation, most SMC nuclei return towards the GMCs within 100 min in control and 5 microM oryzalin treated peels. However, nuclei are unable to reposition in the presence of cytochalasin B (5 micrograms/ml). Thus, the signal for SMC polarisation is issued very early in the ontogeny of the cells (G1), is apparently sustained for a prolonged period, and results in the actin-dependent migration of nuclei towards the GMC. Cytological changes and nuclear migrations similar to those occurring in polarising SMCs can be induced by a local application of pressure to the surface of epidermal cells.

The 25 amino acid residues at the carboxy terminus of the herpes simplex virus type 1 UL26.5 protein are required for the formation of the capsid shell around the scaffold.

Herpes simplex virus type 1 (HSV-1) polypeptides specified by overlapping genes UL26 and UL26.5 form a scaffold around which the icosahedral capsid shell is assembled. In a series of cleavage events catalysed by the UL26-encoded protease, the full-length UL26 product is processed into capsid proteins VP24 and VP21 and the UL26.5 protein is converted into the capsid protein VP22a by the loss of 25 amino acids from its carboxy terminus. The roles of the UL26 and UL26.5 products were investigated using the baculovirus expression system, focusing on the function of the 25 residues cleaved from the UL26.5 protein. A key conclusion from electron microscopic analysis and protein expression studies is that the 25 amino acids at the carboxy terminus of the full-length UL26.5 protein are required for the interaction of the capsid shell proteins with the scaffold in the formation of intermediate capsids. When cells were multiply infected with baculoviruses expressing a truncated form of the UL26.5 product corresponding to VP22a and the essential components of the capsid shell, no capsids were detected, whereas large numbers of capsids were observed when the full-length UL26.5 product was used as a scaffold. The results are consistent with the proposal that cleavage of the UL26.5 product occurs after capsid assembly or when the UL26.5 protein is in a complex with one or more capsid shell proteins. Expression of VP22a in the absence or presence of capsid shell proteins resulted in the formation of large numbers of 60 nm scaffold-like particles. Since VP22a expressed from baculovirus was unable to participate in capsid assembly, these particles cannot be intermediates in the capsid assembly pathway but may be similar in structure to the protein cores present in HSV-1 immature (B) capsids.

Localization of the herpes simplex virus type 1 major capsid protein VP5 to the cell nucleus requires the abundant scaffolding protein VP22a.

The intracellular distributions of three herpes simplex virus type 1 (HSV-1) capsid proteins, VP23, VP5 and VP22a, were examined using vaccinia virus and plasmid expression systems. During infection of cells with HSV-1 wild-type virus, all three proteins were predominantly located in the nucleus, which is the site of capsid assembly. However, when expressed in the absence of any other HSV-1 proteins, although VP22a was found exclusively in the nucleus as expected, VP5 and VP23 were distributed throughout the cell. Thus nuclear localization is not an intrinsic property of these proteins but must be mediated by one or more HSV-1-induced proteins. Co-expression experiments demonstrated that VP5 was efficiently transported to the nucleus in the presence of VP22a, but the distribution of VP23 was unaffected by the presence of either or both of the other two proteins.

Gastrointestinal responses to oats consumption in young adult and elderly rats: digestion, large bowel fermentation and crypt cell proliferation rates.

The present experiment was designed to test the hypothesis that ageing modifies the gastrointestinal responses to a change in diet composition. Rats of the Wag/Rij strain, either young adult (4 months of age) or elderly (27 months of age), were given a basal semi-purified diet or a diet of similar major nutrient composition containing 500 g oatmeal/kg for 17-18 d. Elderly rats digested the dry matter (DM) and organic matter (OM) of both diets less well than did their young adult counterparts, with more of this digestion occurring in the distal intestine. The greater flow of OM to the caecum of oats-fed animals was accompanied by significant reductions in caecal pH and increases in caecal total short-chain fatty acids (SCFA) concentration which appeared to be independent of age. However, young adults responded to feeding on oats by showing a much larger increase in the molar proportion of butyrate (332%) than did elderly animals (79%). Elderly rats had longer duodenal villi than did young adults but effects of age or diet were not detectable at other sites. With both age-groups oats consumption was associated with significant stimulation of crypt cell proliferation rate (CCPR) in the small intestine and caecum, but for the colon there was a significant reduction in CCPR with oats feeding. A reduced ability of the aged large bowel (LB) to produce butyrate may contribute to the prevalence of LB disorders in the elderly.

Frog cones as well as Müller cells have peroxisomes.

We have localized D-amino acid oxidase in peroxisomes of frog retina using cerium procedures on tissue fixed in mixtures containing lower concentrations of glutaraldehyde than we had previously used in our cytochemical studies of this enzyme. We find the Müller cells of these preparations contain a more striking population of peroxisomes than had previously been thought: the D-amino acid oxidase-containing bodies are especially concentrated near the outer limiting membrane, but appreciable numbers are also found in the outer plexiform layer and near the inner limiting membrane. In addition, we find peroxisomes to be present in frog cone photoreceptors, particularly in zones near the ellipsoid. To our knowledge peroxisomes have not been described hitherto in vertebrate photoreceptors. Possible roles for the peroxisomes of the neural retina include participation in the metabolism of lipids (e.g. those of the cones' oil droplets, or of the outer segment) and involvement in oxidation of transmitter-related amino acids and of other small molecules.

Peroxisomal oxidation of thiazolidine carboxylates in firefly fat body, frog retina, and rat liver and kidney.

D-amino acid oxidase is a widely distributed peroxisomal enzyme whose principal natural substrates are still unknown. Thiazolidine carboxylates, their derivatives and relatives, and the intermediates in their metabolism are among the more plausible substrate candidates. Using a cytochemical procedure, we have explored the distribution of peroxide-generating enzymatic activity against two thiazolidine carboxylates. We find that these compounds are effective substrates for peroxisomal oxidation in a variety of tissues that contain peroxisomal D-amino acid oxidase. Reaction was seen in the "classical" peroxisomes of rat liver and kidney, the peroxisomes of the fat body of firefly and of Drosophila and the peroxisomes of frog retina. Interestingly, both with the thiazolidine compounds and with more traditional D-amino acid oxidase substrates, the fireflies' photocyte granules, which are peroxisomes, lack activity.

Peroxisomes in the head of Drosophila melanogaster.

Peroxisomes were localized in the head of wild-type and mutant strains of Drosophila melanogaster by use of a cytochemical method for the demonstration of D-amino acid oxidase activity. With similar techniques we had found previously that vertebrate photoreceptors have few, if any, bodies with cytochemically demonstrable oxidase activity, but that the pigment epithelial cells adjacent to the photoreceptors have a substantial population of such bodies. Peroxisomes in Drosophila were very abundant in the fat body. Probable peroxisomes were also present in the peripheral retina of the eye, including in retinular (retinula) and pigment cells, but there were very few of them. Thus, our results suggest that the fat body, which lies adjacent to the eye, is the principal site of peroxisomal function in the head. Peroxisome functions in the Drosophila head may include participation in the genesis of eye pigments.

The total score of the Crown-Crisp Experiential Index: a useful and valid measure of psychoneurotic pathology.

The Crown-Crisp Experiential Index (CCEI) was designed as a measure of neurotic symptomatology. The tendency has been to report upon its six separate scales and to ignore the total score. We report on the use of the CCEI in a detailed interview study of 208 40-49-year-old women from a local community and 40 patients of the same age range and sex from a psychiatric hospital. In this study, scores on the six scales of the CCEI were shown to be highly intercorrelated. A case is made for combining the six scale scores to form an overall score of neurotic pathology. This score was validated against a number of variables. It was shown to be significantly related to psychiatric patient status, negative self-evaluation, level of severity of current symptoms, evidence of previous psychiatric episodes, previous psychiatric treatment and psychiatric symptoms in childhood. These validation variables were themselves intercorrelated but severity of current symptoms, self-evaluation and childhood symptoms had significant independent effects. It is concluded that scores on the six scales of the CCEI may be less important than the total score which provides a useful and valid measure of psychoneurotic pathology and has particular application as a screening instrument in community surveys.

Life history factors associated with neurotic symptomatology in a rural community sample of 40-49-year-old women.

Associations between a range of life history variables and neurotic pathology (measured by the Crown-Crisp Experiential Index) were examined in a rural community sample of 208 women aged 40-49. Significant childhood associations were a poor maternal relationship, parental discord, nervousness and not liking school. Significant later associations were a poor marital relationship and emotional and behavioural problems in children. There were significant intercorrelations between the significant childhood variables and between the later significant variables but not between the early and later variables. The predictor equations derived from multiple regression analysis fitted significantly a small replication patient sample.

What does the MMPI dependency scale really measure?

Examined the mean DY score of various clinical and nonclinical groups (N = 855). It was significantly higher for females, whether patients or nonpatients; nonpatients with a high total score and high scores on several subscales of the Crown Crisp Experiential Index of neurotic symptomatology; psychiatric patients in general; patients and nonpatients with a high modified Zung depression score; neurotic as opposed to endogenous depressives; suicide attemptors, especially non-serious ones; Ss with early bereavement or separation particularly if followed by poor replacement care; Ss with poor quality marriages and husband dominant marriages as measured by the Ryle Marital Patterns Test. The DY Scale relates directly with the PT and D Scales and inversely with the Do Scale. Separation of the test items into those that were and were not predominantly depressive revealed that most, though not all, significant associations were due to the depressive component, which it was suggested should be markedly diminished.