Neural decoding of visual imagery during sleep.

Visual imagery during sleep has long been a topic of persistent speculation, but its private nature has hampered objective analysis. Here we present a neural decoding approach in which machine-learning models predict the contents of visual imagery during the sleep-onset period, given measured brain activity, by discovering links between human functional magnetic resonance imaging patterns and verbal reports with the assistance of lexical and image databases. Decoding models trained on stimulus-induced brain activity in visual cortical areas showed accurate classification, detection, and identification of contents. Our findings demonstrate that specific visual experience during sleep is represented by brain activity patterns shared by stimulus perception, providing a means to uncover subjective contents of dreaming using objective neural measurement.

Effects of grape seed proanthocyanidins extract on rat mandibular condyle.

We investigated the effects of grape seed proanthocyanidins extract (GSPE) on bone formation by examining total and cortical bone mass, density, architecture, and strength non-invasively using mandibular condyles of Ca-restricted rats. Forty Wistar male rats, each 5 weeks old, were divided into control (C), low-Ca diet (LCaD), low-Ca diet-standard diet (LcaD x SD), and low-Ca diet x Estandard diet with supplementary GSPE (LcaD x SD+GSPE) groups. In LCaD x SD group, after the bone debility was induced by low-Ca diet, a standard diet therapy was given. In LCaD x SD+GSPE group, after the bone debility was induced by low-Ca diet, a standard diet therapy with supplementary GSPE was given. Each mandibular condyle was examined using peripheral quantitative computed tomography (pQCT). There were no significant inter-group differences in body weight seen throughout the experimental period. In LcaD x SD+GSPE, cortical bone cross-sectional area and mineral content were not significantly different from C, while bone mineral content was significantly higher in LcaD x SD+GSPE than in LcaD x SD. Cortical bone density of LcaD x SD+GSPE was not significantly different from that of C, however, that value in LCaD and LcaD x SD was significantly lower than that. The cross-sectional (bending) moment of inertia values in LcaD x SD+GSPE were the highest among all groups, though they did not differ significantly from those in C. Further, the cross-sectional (bending) Stress/Strain Index (SSI) values in LcaD x SD+GSPE were statistically similar to those in C, however, not significant higher than in LcaD x SD. These results suggest that GSPE treatment would increase both bone mass and bone strength on the rat mandibular condyles.

Effects of grape seed proanthocyanidins extract on mandibles in developing rats.

OBJECTIVE: Grape seed proanthocyanidins extract (GSPE), a flavonoid, has a beneficial effect on physical health, which may include the health of bone. The purpose of the present study was to investigate the effects of GSPE on mandibular bone by examining trabecular and cortical bone density, mineral content, and non-invasive bone strength in low-calcium diet rats. MATERIALS AND METHODS: Wistar male rats at 5 weeks old (n = 40) were divided into control (A), low-calcium diet (B), low-calcium diet plus standard diet (C), and low-calcium diet plus standard diet with supplementary GSPE (D) groups. Following 3 weeks of a calcium-restricted diet, group D rats were given 3 mg of GSPE as supplement in 100 g of a standard diet for the next 3 weeks. Following the 6-week experimental period, mandibular bones were examined using peripheral quantitative computed tomography (pQCT). RESULTS: There were no significant differences in body weight or trabecular bone area among the four groups. Trabecular bone density, and trabecular bone mineral content, cortical bone density, cortical bone cross-sectional area, and cortical bone mineral content were significantly higher in group D than in C. Further, Stress-strain index (SSI) values of xSSI and ySSI in group D were significantly higher than in C, although there was no significant difference in pSSI value between those two groups. CONCLUSION: Our results suggest that GSPE treatment caused an increase in both bone formation and bone strength in rat mandibles.

Effects of single-pulse transcranial magnetic stimulation (TMS) on functional brain activity: a combined event-related TMS and evoked potential study.

OBJECTIVE: To further evaluate the potential of slew-rate limiting amplifiers to record electrophysiological signals in spite of concurrent transcranial magnetic stimulation (TMS), and to explore the effects of single-pulse TMS on electroencephalographic (EEG) correlates of functional brain activity. METHODS: Visual-evoked potentials (VEPs) to checkerboards were recorded in 7 right-handed subjects, while single-pulse TMS was applied to the occipital pole either at visual stimulus onset, during the build-up or at the expected peak of the early VEP component P1 (VIS&TMS). Timing of TMS was individually adjusted based on each subject's VEP-latency. A condition of TMS without concurrent visual stimulation (TMS(alone)) served for subtraction purposes (VIS&TMS minus TMS(alone)) to partial out TMS-related contaminations of the EEG signal. RESULTS: When TMS was applied at visual stimulus onset, VEPs (as calculated by subtraction) perfectly matched control VEPs to visual stimulation alone. TMS at around P1, in contrast, modified the targeted (P1) and the subsequent VEP component (N1), independently of whether TMS was given at build-up or peak. CONCLUSIONS: The retrieval of regular VEPs with concomitant TMS at visual stimulus onset suggests that the employed EEG system and subtraction procedure are suited for combined EEG-TMS studies. The VEP changes following TMS at around P1 provide direct clues on the temporal dynamics of TMS pulse effects on functional activity in the human brain. Our data suggest effects of relatively long duration (approximately 100 ms) when TMS is applied while functional neuronal activity evolves.

Sound alters visual evoked potentials in humans.

When a single flash is accompanied by two auditory beeps, the single flash is perceived as two flashes. We investigated whether this crossmodal influence on visual perception occurs at the level of the modality-specific visual pathway or later. We compared the visual evoked potentials (VEPs) in the presence and absence of sound. Activity was modulated extensively and with short latency in trials in which an illusory flash was perceived. In addition, the brain potentials for the illusory flash were qualitatively very similar to those for a physical flash, suggesting that the same mechanism underlies the percept of both illusory and physical flashes. These results suggest that the activity in the visual cortex can be modulated by sound. This implication challenges the general belief that the visual cortical processing is independent of other modalities.

Afterimage of perceptually filled-in surface.

An afterimage induced by prior adaptation to a visual stimulus is believed to be due to bleaching of photochemical pigments or neural adaptation in the retina. We report a type of afterimage that appears to require cortical adaptation. Fixating a neon-color spreading configuration led not only to negative afterimages corresponding to the inducers (local afterimages), but also to one corresponding to the perceptually filled-in surface during adaptation (global afterimage). These afterimages were mutually exclusive, undergoing monocular rivalry. The strength of the global afterimage correlated to a greater extent with perceptual filling-in during adaptation than with the strength of the local afterimages. Thus, global afterimages are not merely by-products of local afterimages, but involve adaptation at a cortical representation of surface.

Identification of an AfsA homologue (BarX) from Streptomyces virginiae as a pleiotropic regulator controlling autoregulator biosynthesis, virginiamycin biosynthesis and virginiamycin M1 resistance.

Virginiae butanolide (VB)-BarA of Streptomyces virginiae is one of the newly discovered pairs of a gamma-butyrolactone autoregulator and the corresponding receptor protein of the Streptomyces species, and has been shown to regulate the production of antibiotic virginiamycin (VM) in S. virginiae. A divergently transcribed barX gene is situated 259 bp upstream of the barA gene, and the BarX protein has been shown to be highly homologous (39.8% identity, 74. 6% similarity) to S. griseus AfsA. Although AfsA is thought to be a biosynthetic enzyme for A-factor, another member of the family of gamma-butyrolactone autoregulators, the in vivo function of S. virginiae BarX was investigated in this study by phenotypic and transcriptional comparison between wild-type S. virginiae and a barX deletion mutant. With the same growth rate as wild-type S. virginiae on both solid and liquid media, the barX mutant showed no apparent changes in its morphological behaviour, indicating that barX does not participate in morphological control in S. virginiae. However, the barX mutant became more sensitive to virginiamycin M1 than did the wild-type strain (minimum inhibitory concentration, 50 microgram ml-1 compared with > 200 microgram ml-1) and exhibited reduced VB and VM production. The VM production was not restored by exogenous addition of VB, suggesting that BarX per se is not a biosynthetic enzyme of VBs but a pleiotropic regulatory protein controlling VB biosynthesis. DNA sequencing of a 5.6 kbp downstream region of barX revealed the presence of five open reading frames (ORFs): barZ, encoding a BarB-like regulatory protein; orf2, encoding a Streptomyces coelicolor RedD-like pathway specific regulator; varM, encoding a homologue of ATP-dependent transporters for macrolide antibiotics; orf4, encoding a homologue of beta-ketoacyl ACP/CoA reductase; and orf5, encoding a homologue of dNDP-glucose dehydratase. Reverse transcription polymerase chain reaction (RT-PCR) analyses of the downstream five genes together with those of the three upstream genes (barA, barB, encoding a regulatory protein; and varS, encoding a virginiamycin S specific transporter) revealed that, in the barX mutant, the transcriptions of barZ, orf2, varM and orf5 were completely repressed and those of barB and varS were derepressed. Because free BarA (BarA in the absence of VB) in wild-type S. virginiae represses the transcription of bicistronic barB-varS operon through binding to a specific DNA sequence (BarA-responsive element, BARE) overlapping the barB transcriptional start site, the derepression of barB-varS transcription in the barX mutant suggested that the in vivo function of BarA was impaired by the lack of BarX protein. Gel-shift assays revealed that BarA easily lost its DNA-binding activity in the absence of BarX but that the defect was restored by the presence of recombinant BarX as a fusion with maltose-binding protein (MBP-BarX), whereas MBP-BarX itself showed no DNA-binding activity, indicating that BarX is likely to be a co-repressor of BarA, enforcing the DNA-binding activity of BarA through protein-protein interactions.

Different recognition by peroxisome proliferator structures in rat peroxisomal induction: application of sandwich ELISA using monoclonal antibody against rat peroxisomes.

A novel assay for a peroxisomal beta-oxidation enzyme by sandwich ELISA using a monoclonal antibody (RPX-5) against purified rat liver peroxisomes was developed. Immunoblot analysis revealed that RPX-5 recognized a 78 Kd protein, which is a peroxisomal bifunctional enzyme (PBE) in the beta-oxidation pathway. Immunoprecipitation by RPX-5 and the resulting reduction of PBE activity were dependent on RPX-5 concentrations. Sandwich ELISA using RPX-5 could be used to assay PBE in the range of 30 to 2000 ng protein/ml. In rat hepatocyte cultures, the PBE amount by this assay correlated well with PBE activity, with correlation coefficients of 0.965. Studying the mechanisms of peroxisomal induction, patterns of peroxisomal induction were examined by co-treatment of rat hepatocytes with various peroxisome proliferators (PxPs). Treatment with clofibrate and bezafibrate resulted in neither an additive nor synergistic effect on PBE level. On the other hand, co-treatment with either bezafibrate-Wy-14,643 or clofibrate-MEHP(mono(2-ethylhexyl)phthalate) both resulted in an additive effect. From these results, it is suggested that PxPs of the fibrate group may exert their functions via a common process, and non-fibrate PxPs via a different process in hepatocytes. The cognition site for peroxisome proliferators, therefore, might not involve a single site for inducing peroxisomal enzymes.

Inhibition by vasoactive intestinal polypeptide (VIP) of angiogenesis induced by murine Colon 26-L5 carcinoma cells metastasized in liver.

We investigated the effect of VIP on the liver metastases and angiogenesis by Colon 26-L5 carcinoma cells in mice. Daily systemic administration of VIP, beginning 3 days after tumor inoculation into a portal vein of mice, inhibited significantly the development of their liver metastases. Immunohistochemical staining for factor VIII-related antigen in the sections of liver metastases showed that the systemic administration of VIP caused significant prevention of angiogenesis within tumor masses. VIP (10-(10) to 10(-6) M) inhibited the invasion of reconstituted basement membrane (Matrigel) by hepatic sinusoidal endothelial (HSE) cells in a concentration-dependent manner in a Transwell chamber assay in vitro and achieved approximately 50% reduction of control at 10(-6) M. VIP (10(-6) M) also significantly suppressed the haptotactic migration of HSE cells to fibronectin, laminin or type I collagen substrates with a similar inhibition rate to the invasion assay. Exposure of VIP to HSE cells induced accumulation of intracellular cAMP in a concentration-dependent manner. The inhibitory effect of VIP (10(-6) M) on HSE cell migration was significantly abrogated in the presence of 3 x 10(-6) M H-89, a cAMP-dependent protein kinase inhibitor. VIP (10(-6) M) inhibited the morphogenesis of HSE cells into capillary-like structures on Matrigel-coated wells. VIP did not affect the proliferation of HSE cells and the production of gelatinases in HSE cells in vitro at the concentrations used in the invasion assay. These observations suggest that the anti-metastatic effect of VIP on liver metastases by Colon 26-L5 carcinoma cells in mice is partly due to the prevention of tumor angiogenesis probably through suppression of the motility of endothelial cells.

Manifestation of scotomas created by transcranial magnetic stimulation of human visual cortex.

Reduced visual performance under transcranial magnetic stimulation (TMS) of human visual cortex demonstrates suppression whose spatial extent is not directly visible. We created an artificial scotoma (region missing from a visual pattern) to directly visualize the location, size and shape of the TMS-induced suppression by following a large-field, patterned, visual stimulus with a magnetic pulse. The scotoma shifted with coil position according to known topography of visual cortex. Visual suppression resulted in pattern-dependent distortion, and the scotoma was filled in with temporally adjacent stimuli, suggesting spatial and temporal completion mechanisms. Thus, perceptual measurements of TMS-induced suppression may provide information about cortical processing via neuronal connections and temporal interactions of neural signals.

Identification and in vivo functional analysis of a virginiamycin S resistance gene (varS) from Streptomyces virginiae.

BarA of Streptomyces virginiae is a specific receptor protein for virginiae butanolide (VB), one of the gamma-butyrolactone autoregulators of the Streptomyces species, and acts as a transcriptional regulator controlling both virginiamycin production and VB biosynthesis. The downstream gene barB, the transcription of which is under the tight control of the VB-BarA system, was found to be transcribed as a polycistronic mRNA with its downstream region, and DNA sequencing revealed a 1,554-bp open reading frame (ORF) beginning at 161 bp downstream of the barB termination codon. The ORF product showed high homology (68 to 73%) to drug efflux proteins having 14 transmembrane segments and was named varS (for S. virginiae antibiotic resistance). Heterologous expression of varS with S. lividans as a host resulted in virginiamycin S-specific resistance, suggesting that varS encoded a virginiamycin S-specific transport protein. Northern blot analysis indicated that the bicistronic transcript of barB-varS appeared 1 to 2 h before the onset of virginiamycin M1 and S production, at which time VB was produced, while exogenously added virginiamycin S apparently induced the monocistronic varS transcript.

Study of chemobiological reactions. 1. Selectivity of aromatic amino compounds and saccharides in glycosylation reactions.

The reactivities between saccharides and primary amino compounds were studied by liquid chromatography on an amino-bonded column and by measurement of the reaction rates of aromatic amines with saccharides. The recoveries from the column and the reaction rates were correlated with their physicochemical properties calculated by the CAChe program. The reactivities between amines and saccharides correlated well with their hydrogen bonding energies calculated by molecular mechanics.

Expression of the Tn antigen on erythroid cells from a patient with Tn syndrome.

In order to examine expression of the Tn antigen on erythroid cells from a patient with Tn syndrome, we applied a selective two phase liquid culture system for human erythroid progenitors in peripheral blood. The cells were analyzed with flow cytometry employing an anti-Tn antibody and a lectin of Vicia villosa which recognizes only the Tn determinant. In the second phase, the Tn antigen was expressed on the cultured cells from the patient on day 3 and Tn-positive cells reached 62.7% on day 9. On the other hand, Tn-positive cells were not detected in the volunteer's cultured cells. When the patient's cells were co-cultured with the cells from a healthy volunteer, the percentage of Tn-positive cells was much lower than the expected value, suggesting that the normal cells suppressed the expression of Tn antigen on the patient's cells.