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Large animal experiments are important for preclinical studies of regenerative stem cell transplantation therapy. Therefore, we investigated the differentiation capacity of pig skeletal muscle-derived stem cells (Sk-MSCs) as an intermediate model between mice and humans for nerve muscle regenerative therapy. Enzymatically extracted cells were obtained from green-fluorescence transgenic micro-mini pigs (GFP-Tg MMP) and sorted as CD34+/45− (Sk-34) and CD34−/45−/29+ (Sk-DN) fractions. The ability to differentiate into skeletal muscle, peripheral nerve, and vascular cell lineages was examined via in vitro cell culture and in vivo cell transplantation into the damaged tibialis anterior muscle and sciatic nerves of nude mice and rats. Protein and mRNA levels were analyzed using RT-PCR, immunohistochemistry, and immunoelectron microscopy. The myogenic potential, which was tested by Pax7 and MyoD expression and the formation of muscle fibers, was higher in Sk-DN cells than in Sk-34 cells but remained weak in the latter. In contrast, the capacity to differentiate into peripheral nerve and vascular cell lineages was significantly stronger in Sk-34 cells. In particular, Sk-DN cells did not engraft to the damaged nerve, whereas Sk-34 cells showed active engraftment and differentiation into perineurial/endoneurial cells, endothelial cells, and vascular smooth muscle cells, similar to the human case, as previously reported. Therefore, we concluded that Sk-34 and Sk-DN cells in pigs are closer to those in humans than to those in mice. [ABSTRACT FROM AUTHOR]

Introduction An essential complement to molecular-genetic approaches for analyzing the function of the oculomotor circuitry in mice is an understanding of sensory and motor signal processing in the circuit. Although there has been extensive analysis of the signals carried by neurons in the oculomotor circuits of species, such as monkeys, rabbits and goldfish, relatively little in vivo physiology has been done in the oculomotor circuitry of mice. We analyzed the contribution of vestibular and nonvestibular signals to the responses of individual Purkinje cells in the cerebellar flocculus of mice. Methods We recorded Purkinje cells in the cerebellar flocculus of C57 BL/6 mice during eye movement responses to vestibular and visual stimulation. Results As in other species, most individual Purkinje cells in mice carried both vestibular and nonvestibular signals, and the most common response across cells was an increase in firing in response to ipsiversive eye movement or ipsiversive head movement. When both the head and eyes were moving, the Purkinje cell responses were approximated as a linear summation of head and eye velocity inputs. Unlike other species, floccular Purkinje cells in mice were considerably more sensitive to eye velocity than head velocity. Conclusions The signal content of Purkinje cells in the cerebellar flocculus of mice was qualitatively similar to that in other species. However, the eye velocity sensitivity was higher than in other species, which may reflect a tuning to the smaller range of eye velocities in mice. [ABSTRACT FROM AUTHOR]

The commercial quality of Japanese Angelica radices - Angelica acutiloba Kitagawa (Yamato-toki) and A. acutiloba Kitagawa var. sugiyama Hikino (Hokkai-toki) - used in Kampo traditional herbal medicines, was studied by use of omics technologies. Complementary and alternative medical providers have observed in their clinical experience that differences in radix commercial quality reflect the differences in pharmacological responses; however, there has been little scientific examination of this phenomenon. The approach of omics, including metabolomics, transcriptomics, genomics, and informatics revealed a distinction between the radix-quality grades based on their metabolites, gene expression in human subjects, and plant genome sequences. Systems biology, constructing a network of omics data used to analyze this complex system, is expected to be a powerful tool for enhancing the study of radix quality and furthering a comprehensive understanding of all medicinal plants. [ABSTRACT FROM AUTHOR]

The commercial quality of Japanese Angelica radices - Angelica acutiloba Kitagawa (Yamato-toki) and A. acutiloba Kitagawa var. sugiyama Hikino (Hokkai-toki) - used in Kampo traditional herbal medicines, was studied by use of omics technologies. Complementary and alternative medical providers have observed in their clinical experience that differences in radix commercial quality reflect the differences in pharmacological responses; however, there has been little scientific examination of this phenomenon. The approach of omics, including metabolomics, transcriptomics, genomics, and informatics revealed a distinction between the radix-quality grades based on their metabolites, gene expression in human subjects, and plant genome sequences. Systems biology, constructing a network of omics data used to analyze this complex system, is expected to be a powerful tool for enhancing the study of radix quality and furthering a comprehensive understanding of all medicinal plants. [ABSTRACT FROM AUTHOR]

Aim of the study: Japanese Angelica radix (Angelica acutiloba Kitagawa radix; Yamato-toki), is classified into one of the three grades: excellent-, middle- and low-grade. Even though herbal doctors and Kampo pharmacists have observed that radix grades correspond to their pharmacological response in clinical experience, these observations has not been documented with supporting scientific evidence in the literature. In the current study, relationship between content of pharmacological components and the grade of radixes was reported. Materials and methods: Pharmacological effect of methanol extracts of excellent- and low-grade radixes was compared using assay of inhibition of acetylcholine-induced contraction in guinea pig ileum. Pharmacological contents of the radix were quantitatively measured by HPLC analysis. Results: Excellent-grade radixes showed lower activity than low-grade radixes on the inhibition assay for ileum contraction. (Z)-Ligustilide content of the excellent-grade radixes was statistically significantly lower than that of other grades of radixes (p-value by Student''s t-test: 6.9×10−4). A regression equation was obtained from assessing the inhibitory activity of a (Z)-ligustilide standard at various concentrations. Comparison of the inhibitory activity values, which were estimated by the regression equation, and the actual activity values, which were determined using the assay, indicated that ligustilide is the factor that dominantly affects the activity. The ligustilide concentration of hot water extracts of radixes was determined. The concentration in the hot water extracts and the content of the radixes was positively correlated based on a Pearson correlation test. This result suggests that ligustilide extracted by infusion depends on its radix content. Conclusions: There are differences in (Z)-ligustilide content and its pharmacological activity depending on the grade of Japanese Angelica radix. [Copyright &y& Elsevier]

Lophine hydroperoxides underwent base-triggered 1,5-phenyl migration in DMSO to afford imidazolones in high yields, instead of amidines with chemiluminescence (CL). The corresponding imidazolols were believed to intermediates and they were successfully obtained by treating the peroxides with DMSO without the base. The diminished CL was because of the reduction of the hydroperoxides with DMSO. The imidazolols subsequently underwent smooth base-mediated rearrangement to afford imidazolones. Furthermore, the chiral imidazolols provided stereoselective imidazolones in high enantiomeric excess (>92%), which supported the mechanism of an intramolecular ring for the migration. J. Heterocyclic Chem., (2010). [ABSTRACT FROM AUTHOR]

Abstract: Vanadyl sulfate (VOSO4) has been clinically tested in diabetic patients since 1995. Oral administrations of VOSO4 improved the type 2 diabetic state with respect to plasma glucose, HbA1c, and fructosamine levels. The development of toxicity by increasing the administration of VOSO4 should be avoided. One method was the utilization of vanadyl complexes with coordination compounds that are low-toxic and low-molecular-weight ligands to enhance the permeation of the metal ion to lipid bilayer membrane. Over a decade we have focused on a variety of heterocyclic compounds as bidentate ligands for metal ions. Vanadyl and zinc(II) complexes of 1-substituted 3-hydroxy-2-methyl-4(1H)-pyridinethiones, 4,5,6-substituted 1-hydroxy-2(1H)-pyrimidinones, 4-(p-substituted)phenyl-3-hydroxythiazole-2(3H)-thiones, 3-hydroxypyrone, 1-alkyl- or 1-phenylalkyl-3-hydroxy-2(1H)-pyridinethiones, optically active 1-substituted 3-hydroxy-4(1H)-pyridinethiones, and 5-dialkylsulfonamido- or 5,7-bis(dialkylsulfonamido)-8-hydroxyquinolines were prepared, and their insulin-mimetic activities were evaluated in terms of IC50 values which stand for a 50% inhibitory concentration of the free fatty acid release from isolated rat adipocytes. In this article, the relationship between the insulin-mimetic activity and the partition coefficient, the chirality, the substituent effect, molecular weight, the pK a value, and the coordination mode was discussed. In vivo blood glucose-lowering effects of the vanadyl complex with 1-hydroxy-4,6-dimethyl-2(1H)-pyrimidinone in streptozotocin (STZ)-induced diabetic rats and the zinc(II) complexes with 4-(p-chlorophenyl)thiazole- and 4-methylthiazole-2(3H)-thione in KK-Ay mice were also discussed. [Copyright &y& Elsevier]

To optimize motor performance, both the amplitude and temporal properties of movements should be modifiable by motor learning. Here we report that the modification of movement timing is highly dependent on signaling through P/Qtype voltage-dependent calcium channels. Two lines of mutant mice heterozygous for P/Q-type voltage-dependent calcium channels exhibited impaired plasticity of eye movement timing, but relatively intact plasticity of movement amplitude during motor learning in the vestibulo-ocular reflex. The results thus demonstrate a distinction between the molecular signaling pathways regulating the timing versus amplitude of movements. [ABSTRACT FROM AUTHOR]

Abstract: The prices of Angelica acutiloba Kitagawa (yamato-toki) and A. acutiloba Kitagawa var. sukiyamae Hikino (hokkai-toki) are now mainly determined according to the sensory quality determined by experts in addition to the physical properties. This method provides a low reliability result for differentiating and qualifying their qualities. In addition, the quality in terms of pharmacological efficiency is not taken into account for consideration in the ordinary sensory method. A combination of a 1H NMR technique and a multivariate analysis was preliminarily applied for the quality evaluation of both toki roots with regard to their geographical and variety differences. A broad range of metabolites was detected by a single-run 1H NMR spectrometry. Partial least-squares discrimination analysis (PLS-DA), a pattern recognition method, was applied to the 1H NMR spectra of aqueous extracts of toki samples having different sensory qualities. The PLS-DA result showed a clear clustering corresponding to the cultivation area between toki samples cultivated in Hokkaido (Japan) and those cultivated in the southern part of China and the Nara prefecture (Japan), while there was no separation corresponding to the toki''s variety and sensory qualities, indicating the inconsistency of the sensory evaluation result. The chemical metabolites contributing to the discrimination of toki samples in relation to pharmacological and sensory properties were reported for the first time. A reliable multivariate calibration model used to predict the sensory quality was successfully carried out by PLS regression. [Copyright &y& Elsevier]

Abstract: Two fluorescent monomers having a quinoxaline skeleton, N-(2,3-dimorpholinoquinoxalin-6-yl)acrylamide (QxA) and N-(1-(2,3-dimorpholinoquinoxalin-6-ylamino)prop-2-yl)methacrylamide (QxAlaMA), were synthesized. Thermo-responsive copolymers of N-isopropylacrylamide (NIPAM) and a small amount of a fluorescent monomer were synthesized and their fluorescence properties investigated. The fluorescent monomers showed intense solvatochromism in their fluorescence. The wavelength at the maximum fluorescence intensity of the QxAlaMA-labeled PNIPAM dramatically blue-shifted and the fluorescence intensity of the QxA-labeled PNIPAM significantly increased around the transition temperature. It was found that these fluorescent dyes can sense and report the thermo-responsive behavior of the PNIPAM in water. Both QxAlaMA and QxA were demonstrated to be applicable to new intramolecular fluorescent probes. [Copyright &y& Elsevier]

Summary: The number and diversity of plasticity mechanisms in the brain raises a central question: does a neural circuit store all memories by stereotyped application of the available plasticity mechanisms, or can subsets of these mechanisms be selectively engaged for specific memories? The uniform architecture of the cerebellum has inspired the idea that plasticity mechanisms like cerebellar long-term depression (LTD) contribute universally to memory storage. To test this idea, we investigated a set of closely related, cerebellum-dependent motor memories. In mutant mice lacking Ca2+/calmodulin-dependent protein kinase IV (CaMKIV), the maintenance of cerebellar LTD is abolished. Although memory for an increase in the gain of the vestibulo-ocular reflex (VOR) induced with high-frequency stimuli was impaired in these mice, memories for decreases in VOR gain and increases in gain induced with low-frequency stimuli were intact. Thus, a particular plasticity mechanism need not support all cerebellum-dependent memories, but can be engaged selectively according to the parameters of training. [Copyright &y& Elsevier]

NAD is a ubiquitous coenzyme involved in oxidation-reduction reactions and is synthesized by way of quinolinate. Animals and some bacteria synthesize quinolinate from tryptophan, whereas other bacteria synthesize quinolinate from aspartate (Asp) using L-Asp oxidase and quinolinate synthase. We show here that Arabidopsis (Arabidopsis thaliana) uses the Asp-to-quinolinate pathway. The Arabidopsis L-Asp oxidase or quinolinate synthase gene complemented the Escherichia coli mutant defective in the corresponding gene, and T-DNA-based disruption of either of these genes, as well as of the gene coding for the enzyme quinolinate phosphoribosyltransferase, was embryo lethal. An analysis of functional green fluorescent protein-fused constructs and in vitro assays of uptake into isolated chloroplasts demonstrated that these three enzymes are located in the plastid. [ABSTRACT FROM AUTHOR]

Two kinds of 3-hydroxypyridine-2(1 H )-thiones were synthesized. The visible (VIS) spectroscopic analysis indicated that 3-hydroxy-1-methylpyridine-2(1 H )-thione (4a) and 3-hydroxy-1-(2-hydroxyethyl)pyridine-2(1 H )-thione (4b) formed stable 3:1 Fe(III) complexes. The stability constant of the 4b-Fe(III) complex was estimated from the competitive reaction with EDTA and was found to be 36.7 in logß 3 . Treatment of compound 4b with Ga(acac) 3 in D 2 O:CD 3 OD (9:1) solution afforded 3:1 Ga(III) complex, which was assigned by means of 1 H nuclear magnetic resonance ( 1 H NMR) spectroscopy. Treatment of compound 4b with Pb(NO 3 ) 2 gave 4b-Pb(II) complex. The Pb(II) selectivity over biologically relevant Mg(II) and Ca(II) was remarkably improved by adopting N -hydroxyethyl functionality instead of N -methyl group. [ABSTRACT FROM AUTHOR]

The ionotropic glutamate receptorδ2 subunit (GluRδ2) is selectively expressed in cerebellar Purkinje cells and is implicated in long-term depression, synaptic formation and elimination. To study the effect of GluRδ2 deficiency on motor control, we measured the vestibulo-ocular reflex (VOR) and optokinetic response (OKR) induced by sinusoidal rotation of the animal and/or the surrounding screen in two GluRδ2 mutant mice: a GluRδ2 knockout mouse (δ2–/–) and alurchermouse with a point mutation in the GluRδ2 gene resulting in loss of all Purkinje cells.δ2–/– showed significantly higher VOR gain in the dark (VORD) than in the wild-type. Inδ2–/–, the VOR gain in light was lower than that in the dark. The phase of OKR lagged more inδ2–/– than inlurcherand wild-type mice. Both mutant mice failed to change the VORD or OKR gain adaptively in response to sustained vestibular and/or visual stimulation. Basal properties of VOR and OKR changed little by lesion of the flocculus, but they changed substantially by lesion of the inferior olivary nuclei (IO). The abnormal VOR gain and OKR phase delay were clearly reduced inδ2–/– by the latter lesion. Our results indicate that failures in the GluRδ2-dependent synaptic regulation affect motor performance more severely than loss of cerebellar cortical outputs. This study suggests that the anomalies inδ2–/– are dependent on inputs from IO and that GluRδ2 deficiency changed properties of not only the cerebellar cortex but also the brainstem neuronal pathways controlling reflex eye movements during development. [ABSTRACT FROM AUTHOR]

The cerebellum is an evolutionarily conserved structure critical for motor learning in vertebrates. The model that has influenced much of the work in the field for the past 30 years suggests that motor learning is mediated by a single plasticity mechanism in the cerebellum: long-term depression (LTD) of parallel fiber synapses onto Purkinje cells. However, recent studies of simple behaviors such as the vestibulo-ocular reflex (VOR) indicate that multiple plasticity mechanisms contribute to cerebellum-dependent learning. Multiple plasticity mechanisms may provide the flexibility required to store memories over different timescales, regulate the dynamics of movement, and allow bidirectional changes in movement amplitude. These plasticity mechanisms must act in combination with appropriate information-coding strategies to equip motor-learning systems with the ability to express learning in correct contexts. Studies of the patterns of generalization of motor learning in the VOR provide insight about the coding of information in neurons at sites of plasticity. These principles emerging from studies of the VOR are consistent with results concerning more complex behaviors and thus may reflect general principles of cerebellar function. [ABSTRACT FROM AUTHOR]

How failures in regulation of synaptic transmission in the mammalian CNS affect neuronal activity and disturb motor coordination is addressed. The mutant mouse deficient in the glutamate receptor δ2 subunit, specifically expressed in cerebellar Purkinje neurons, has defects in synaptic regulations such as synaptic plasticity, stabilization, and elimination of synaptic connections and shows failures in motor coordination and learning. In this study, the cause of motor discoordination of the δ2 mutant mouse was analyzed by comparing its motor control ability with those of the wild-type mouse and the lurcher mutant mouse, which loses all Purkinje neurons, the sole output neurons in the cerebellar cortex. Unexpectedly, the δ2 mutant mouse showed severer motor discoordination than the lurcher mouse without any cerebellar cortical outputs. The δ2 mutant mouse showed involuntary spontaneous eye movement with characteristic 10 Hz oscillation, which disappeared by ablation of the cerebellar flocculus, suggesting that the δ2 mutant cerebellar cortex outputs an abnormal signal. In vivo extracellular recordings of neuronal activity revealed that Purkinje neurons tended to fire clustered action potentials and complex spikes at ∼ 10 Hz in the δ2 mutant mouse. A whole-cell patch-damp recording from Purkinje neurons in cerebellar slices indicated that the clustered action potentials could be induced by climbing fiber activation. Taken together, our results suggest that the δ2 subunit deficiency produces the oscillating activity in Purkinje neurons by enhancing climbing fiber inputs, causing surplus movement and affecting motor control worse than no signal at all. [ABSTRACT FROM AUTHOR]

Metabotropic glutamate receptor subtype 1 (mGluR1) plays an essential role in the cerebellar long-term depression (LTD). We examined the dynamic characteristics and adaptability of horizontal vestibulo-ocular reflex (HVOR) and optokinetic response (HOKR) eye movements in mGluR1 knockout mice. A mild difference was seen in the HOKR/HVOR dynamics between the wild-type and mGluR1(−/−) mice. Exposure to 1 h of sustained screen oscillation, which induced HOKR adaptation in wild-type mice, induced no change in mutant mice. These results suggest that the mGluR1 plays an essential role in the adaptation of HOKR, and LTD underlies the adaptation of ocular reflexes. [Copyright &y& Elsevier]

Describes the synthesis of novel hexadentate ligands composed of N-hydroxy-2(1H)-pyrazinone, amino acids, and tris(2-aminoethyl)amine. FeIII-chelating properties; Ultraviolet-visible spectral evidence of formation of intramolecular 1:3 FeIII complex to the hydroxamate units; Relative stability constants of the complexes.

Develops a mutant of the Synechocystis sp. strain PCC6803 where the cotA gene was completed deleted. CotA gene as a homolog of the cemA genes found in chloroplast genomes of higher plants; Evidence that this mutant does not extrude protons in the light; Possible role of cotA in light-induced proton extrusion.

Cites a study which measured the characteristics of reflex eye movements, using an infrared television camera system. What the reflex eye movement in mice showed; Inforamtion on the evaluation of dynamic characteristics of eye movement in mice; In-depth look at the results of the study.

A cobalt(III) complex with chiral ligand, H2cpel (N-carboxymethylN-pyridylethyl-L-leucine), was prepared for chiral recognition of amino acids. Through the competitive coordination of racemic phenylalanine to the chiral cobalt complex, [Co(cpel)(CO[sub 3])][sup -] (1), enantioselective recognition was achieved on the ternary complex, which was determined on the basis of HPLC analysis with a chiral column. The formation rate for the [Co(cpel)(L-phe)] complex (2) was 6-times superior to that of [Co(cpel)(D-phe)] (3). The preferential formation of 2 might be illustrated by the interligand π-π stacking interaction. Crystal structural analysis for 2 and 3 revealed that aromatic rings, pyridine ring of CPEL and phenylalanine sidechain, in 2 were very close each other but those in 3 were far apart. Such interligand aromatic interaction in 2 was also examined by the use of ¹H NMR spectra. [ABSTRACT FROM AUTHOR]

The smooth pursuit system has the ability to perform predictive feedforward control of eye movements. This study attempted to examine how stimulus and behavioral histories of past trials affect the control of predictive pursuit of target motion with randomized velocities. We used sequential ramp stimuli where the rightward velocity was fixed at 16 deg/s while the leftward velocity was either fixed (predictable) at one of seven velocities (4, 8, 12, 16, 20, 24, or 28 deg/s) or randomized (unpredictable). As a result, predictive pursuit responses were observed not only in the predictable condition but also in the unpredictable condition. Linear mixed-effects (LME) models showed that both stimulus and behavioral histories of the previous two or three trials influenced the predictive pursuit responses in the unpredictable condition. Intriguingly, the goodness of fit of the LME model was improved when both historical effects were fitted simultaneously rather than when each type of historical data was fitted alone. Our results suggest that predictive pursuit systems allow us to track randomized target motion using weighted averaging of the information of target velocity (stimulus) and motor output (behavior) in past time sequences. [ABSTRACT FROM AUTHOR]

Summary Long-term depression (LTD) of AMPA-type glutamate receptor (AMPA receptor)-mediated synaptic transmission has been proposed as a cellular substrate for learning and memory. Although activity-induced AMPA receptor endocytosis is believed to underlie LTD, it remains largely unclear whether LTD and AMPA receptor endocytosis at specific synapses are causally linked to learning and memory in vivo. Here we developed a new optogenetic tool, termed PhotonSABER, which enabled the temporal, spatial, and cell-type-specific control of AMPA receptor endocytosis at active synapses, while the basal synaptic properties and other forms of synaptic plasticity were unaffected. We found that fiberoptic illumination to Purkinje cells expressing PhotonSABER in vivo inhibited cerebellar motor learning during adaptation of the horizontal optokinetic response and vestibulo-ocular reflex, as well as synaptic AMPA receptor decrease in the flocculus. Our results demonstrate that LTD and AMPA receptor endocytosis at specific neuronal circuits were directly responsible for motor learning in vivo. Video Abstract Graphical Abstract Highlights • A new optogenetic tool, PhotonSABER, controls endocytosis at active synapses • PhotonSABER inhibits AMPA receptor endocytosis during LTD in a light-dependent manner • Basal synaptic transmission and other forms of synaptic plasticity were unaffected • PhotonSABER inhibits AMPA receptor endocytosis and motor learning in Purkinje cells in vivo Kakegawa et al. show that AMPA receptor endocytosis at parallel fiber-Purkinje cell synapses in the cerebellar flocculus plays a direct role in motor learning in vivo , using a new optogenetic tool, PhotonSABER, which enables the acute inhibition of AMPA receptor endocytosis during LTD. [ABSTRACT FROM AUTHOR]

Abstract Among the subtypes of the Ca2+ -dependent protein kinase C (PKC), which play a crucial role in long-term depression (LTD), both α and γ are expressed in the cerebellar floccular Purkinje cells. To reveal the functional differences of PKC subtypes, we examined the adaptability of ocular reflexes of PKCγ mutant mice, which show mild ataxia and normal LTD. In mutant mice, gains of the horizontal optokinetic eye response (HOKR) were reduced. Adaptation of the HOKR was not affected but its retinal slip dependency was altered in mutant mice. Sustained 1-h sinusoidal screen oscillation, which induced a relatively large amount of retinal slips in both mutant and wild-type mice, increased the HOKR gain in wild-type mice but not in mutant mice. In contrast, exposure to 1 h of sustained slower screen oscillations, which induced relatively small retinal slips in mutant and wild-type mice, increased the HOKR gain in both mutant and wild-type mice. Adaptation of the HOKR of the mutant mice to slow screen oscillation and those of wild-type mice to fast and slow screen oscillations were all abolished by local applications of a PKC inhibitor (chelerythrine) within the flocculi. Electrophysiological and anatomical studies showed no appreciable changes in the sources and magnitudes of climbing fibre inputs, which mediate retinal slip signals to the flocculus in the mutant mice. These results suggest that PKCγ has a modulatory role in determining retinal slip dependency, and other PKC subtypes, e.g. PKCα, may play a crucial role in the adaptation of the HOKR. [ABSTRACT FROM AUTHOR]

The DHCR7 enzyme converts 7-DHC into cholesterol. Mutations in DHCR7 can block cholesterol production, leading to abnormal accumulation of 7-DHC and causing Smith–Lemli–Opitz syndrome (SLOS). SLOS is an autosomal recessive disorder characterized by multiple malformations, including microcephaly, intellectual disability, behavior reminiscent of autism, sleep disturbances, and attention-deficit/hyperactivity disorder (ADHD)-like hyperactivity. Although 7-DHC affects neuronal differentiation in ex vivo experiments, the precise mechanism of SLOS remains unclear. We generated Dhcr7 deficient (dhcr7 −/− ) zebrafish that exhibited key features of SLOS, including microcephaly, decreased neural stem cell pools, and behavioral phenotypes similar to those of ADHD-like hyperactivity. These zebrafish demonstrated compromised myelination, synaptic anomalies, and neurotransmitter imbalances. The axons of the dhcr7 −/− zebrafish showed increased lysosomes and attenuated autophagy, suggesting that autophagy-related neuronal homeostasis is disrupted. • DHCR7 mutations lead to SLOS due to disrupted cholesterol homeostasis. • Cholesterol decreases, whereas 7-DHC increases in dhcr7 −/− zebrafish. • Dhcr7 −/− zebrafish show microcephaly, synaptic defects, and ADHD-like hyperactivity. • DHCR7 deficiency disrupts autophagy-related neuronal homeostasis in axons. [ABSTRACT FROM AUTHOR]

A total of 42 trisubstituted carboranes categorised into five scaffolds were systematically designed and synthesized by exploiting the different reactivities of the twelve vertices of o-, m-, and p-carboranes to cover all directions in chemical space. Significant inhibitors of hypoxia inducible factor transcriptional activitay were mainly observed among scaffold V compounds (e.g., Vi–m, and Vo), whereas anti-rabies virus activity was observed among scaffold V (Va–h), scaffold II (IIb–g), and scaffold IV (IVb) compounds. The pharmacophore model predicted from compounds with scaffold V, which exhibited significant anti-rabies virus activity, agreed well with compounds IIb–g with scaffold II and compound IVb with scaffold IV. Normalized principal moment of inertia analysis indicated that carboranes with scaffolds I–V cover all regions in the chemical space. Furthermore, the first compounds shown to stimulate the proliferation of the rabies virus were found among scaffold V carboranes. [ABSTRACT FROM AUTHOR]

In this study, the ability to tailor the peptide-binding specificity of an RNA was investigated. First, variants of the Rev-response element (RRE) RNA with different specificities toward the natural binding partner, Rev, and two RRE-binding aptamers, the RSG-1.2 and the Kl peptides, were identified. Next, hybrid RRE mutants with combinations of two sets of specificity-altering substitutions were tested for peptide-binding specificity. It was shown that in most cases the results of the combination of individual mutations were of an additive nature, therefore providing a way to manipulate the peptide-binding specificity of an RNA in a predictable manner. [ABSTRACT FROM AUTHOR]

The antidiabetic effect of vanadium is a widely accepted phenomenon; some oxovanadium(IV) complexes have been found to normalize high blood glucose levels in both type 1 and type 2 diabetic animals. In light of the future clinical use of these complexes, the relationship among their chemical structures, physicochemical properties, metallokinetics, and antidiabetic activities must be closely investigated. Recently, we found that among bis(3-hydroxypyronato)oxovanadium(IV) [VO(3hp)2] related complexes, bis(allixinato)oxovanadium(IV) [VO(alx)2] exhibits a relatively strong hypoglycemic effect in diabetic animals. Next, we examined its metallokinetics in the blood of rats that received five VO(3hp)2-related complexes by the blood circulation monitoring–electron paramagnetic resonance method. The metallokinetic parameters were obtained from the blood clearance curves based on a two-compartment model; most parameters, such as area under the concentration curve and mean residence time, correlated significantly with the in vitro insulinomimetic activity in terms of 1/IC50 (IC50 is the 50% inhibitory concentration of the complex required for the release of free fatty acids in adipocytes) and the lipophilicity of the complex (log P com). The oxovanadium(IV) concentration was significantly higher and the species resided longer in the blood of rats that received VO(alx)2 than in the blood of rats that received VO(3hp)2 or bis(kojato)oxovanadium(IV); VO(alx)2 also exhibited higher log P com and 1/IC50 values. On the basis of these results, we propose that the introduction of lipophilic groups at the C2 and C6 positions of the 3hp ligand is an effective method to enhance the hypoglycemic effect of the complexes, as supported by the observed in vivo exposure and residence in the blood. [ABSTRACT FROM AUTHOR]

A novel sp3 carbon‐rich tricyclic 3D scaffold‐based peptide mimetic compound library was constructed to target protein‐protein interactions. Tricyclic framework 7 was synthesized from 9‐azabicyclo[3,3,1]nonan‐3‐one (11) via a gold(I)‐catalyzed Conia‐ene reaction. The electron‐donating group on the pendant alkyne of cyclization precursor 12 b–e was the key to forming 6‐endo‐dig cyclized product 7 with complete regioselectivity. Using the synthetic strategy for regioselective construction of bridged tricyclic framework 7, a diazatricyclododecene 3D‐scaffold 8 a, which enables the introduction of substituents into the scaffold to mimic amino acid side chains, was designed and synthesized. The peptide mimetics 21 a–u were synthesized via step‐by‐step installation of three substituents on diazatricyclododecene scaffold 8 a. Compounds 21 a–h were synthesized as α‐helix peptide mimics of hydrophobic ZZxxZ and ZxxZZ sequences (Z=Leu or Phe) and subjected to cell‐based assays: antiproliferative activity, HIF‐1 transcriptional activity which is considered to affect cancer malignancy, and antiviral activity against rabies virus. Compound 21 a showed the strongest inhibitory activity of HIF‐1 transcriptional activity (IC50=4.1±0.8 μM), whereas compounds 21 a–g showed antiviral activity with IC50 values of 4.2–12.4 μM, suggesting that the 3D‐scaffold 8 a has potential as a versatile peptide mimic skeleton. [ABSTRACT FROM AUTHOR]

The purpose of this study was to determine whether smooth pursuit eye movements affect visual motion prediction using a time‐to‐contact task where observers anticipate the exact instant that a partially occluded target would coincide with a stationary object. Moreover, we attempted to clarify the influence of second‐order motion on visual motion prediction during smooth pursuit. One target object moved to another stationary object (6 deg apart) at constant velocity of 3, 4, and 5 deg/s, and then the two objects disappeared 500 ms after the onset of target motion. The observers estimated the moment the moving object would overlap the stationary object and pressed a button. For the pursuit condition, both a Gaussian window and a random dots texture moved in the same direction at the same speed for the first‐order motion, whereas a Gaussian window moved over a static background composed of random dots texture for the second‐order motion. The results showed that the constant error of the time‐to‐contact shifted to a later response for the pursuit condition compared to the fixation condition, regardless of the object velocity. In addition, during smooth pursuit, the constant error for the second‐order motion shifted to an earlier response compared to the first‐order motion when the object velocity was 3 deg/s, whereas no significant difference was found at 4 and 5 deg/s. Therefore, our results suggest that visual motion prediction using a time‐to‐contact task is affected by both eye movements and motion configuration such as second‐order motion. [ABSTRACT FROM AUTHOR]

Endometriosis exhibits unique characteristics, such as fibrosis, resistance to apoptosis, and promotion of cell proliferation; however, its pathophysiology is not fully understood. Recurrence rates after treatment are high, and the progression risk continues until menopause; hence, more effective therapy for endometriosis is needed. CREB-binding protein (CBP)/β-catenin signaling inhibitors have demonstrated antifibrogenetic effects in liver, lung, and skin diseases. The present study evaluated the effects of two CBP/β-catenin signaling inhibitors, ICG-001 and C-82, on the progression of endometriosis using endometriotic cyst stromal cells from the ovary and normal endometrial stromal cells from the uterus. ICG-001 was also evaluated in a mouse model. ICG-001 and C-82 inhibited cell proliferation, fibrogenesis, and cell migration, and promoted apoptosis in vitro. ICG-001 inhibited the growth of endometriotic lesions in the mouse model. CBP/β-catenin signaling plays an important role in the pathophysiology of endometriosis. Inhibiting the CBP/β-catenin signal can be a therapeutic target for endometriosis. [ABSTRACT FROM AUTHOR]

Enhanced root growth is known as the survival strategy of plants under drought. Previous proteome analysis in drought-resistant wild watermelon has shown that Ran GTPase, an essential regulator of cell division and proliferation, was induced in the roots under drought. In this study, two cDNAs were isolated from wild watermelon,CLRan1andCLRan2, which showed a high degree of structural similarity with those of other plant Ran GTPases. Quantitative RT-PCR and promoter-GUS assays suggested thatCLRan1was expressed mainly in the root apex and lateral root primordia, whereasCLRan2was more broadly expressed in other part of the roots. Immunoblotting analysis confirmed that the abundance of CLRan proteins was elevated in the root apex region under drought stress. Transgenic Arabidopsis overexpressing CLRan1 showed enhanced primary root growth, and the growth was maintained under osmotic stress, indicating thatCLRan1functions as a positive factor for maintaining root growth under stress conditions. Ran GTPase (CLRan1), which is expressed in the roots of drought-resistant wild watermelon, functions as a positive factor for maintaining root growth under osmotic stress. [ABSTRACT FROM PUBLISHER]

The effects of bicarbonate and its combination with the chelating agents, deferiprone (L1), 4,6-dimethyl-1-hydroxypyrimidin-2(1H)-one (AK-4), catechol-3,6-bis(methyleneimino-diacetic-acid) (CBMIDA), and ethane-1-hydroxy-1,1-bisphoshonate (EHBP) in removing depleted uranium (DU) for radiation emergency medicine were examined. After the intramuscular injection of DU in rats, various time schedules of bicarbonate and chelating agent administration were tested. The results indicate that the bicarbonate helps increase significantly the effects of LI and AK-4, while there were no effects of using bicarbonate alone. The effects of bicarbonate on CBMIDA were unclear, and the effects of EHBP were negative. Further studies are necessary to obtain distinctly synergic effects by the combination of chelating agents with bicarbonate. [ABSTRACT FROM AUTHOR]

Abstract: Previously, we found that bis(allixinato)oxovanadium(IV) (VO(alx)2) exhibits a potent hypoglycemic activity in type 1-like diabetic mice. Since the enhancement of insulin sensitivity is involved in one of the mechanisms by which vanadium exerts its anti-diabetic effects, VO(alx)2 was further tested in type 2 diabetes with low insulin sensitivity. The effect of oral administration of VO(alx)2 was examined in obesity-linked type 2 diabetic KKAy mice. Treatment of VO(alx)2 for 4 weeks normalized hyperglycemia, glucose intolerance, hyperinsulinemia, hypercholesterolemia and hypertension in KKAy mice; however, it had no effect on hypoadiponectinemia. VO(alx)2 also improved hyperleptinemia, following attenuation of obesity in KKAy mice. This is the first example in which a vanadium compound improved leptin resistance in type 2 diabetes by oral administration. On the basis of these results, VO(alx)2 is proposed to enhance not only insulin sensitivity but also leptin sensitivity, which in turn improves diabetes, obesity and hypertension in an obesity-linked type 2 diabetic animal. [Copyright &y& Elsevier]

The effectiveness of deferiprone (L1) in removing depleted uranium (DU) and protecting animals from radiation exposure was examined. Rats that had received 2 mg/kg DU via intramuscular injection were orally administered 100, 200 or 400 mg/kg L1 for 3 days. In all of the groups, significant increases in urinary DU excretion and decreases in DU concentration in the injected muscle were observed, indicating that L1 combined with DU and DU was excreted in the urine. No significant increase in the amount of DU in the excreta or decrease in DU concentration in organs other than the muscles was found. As a preliminary test, the effectiveness of L1 in reducing radiation damage was examined in mice injected with 400 mg/kg L1 and rats administered orally with 200 and 400 mg/kg L1 before and just after x-ray exposure. The results were inconclusive. [ABSTRACT FROM AUTHOR]

Abstract: Control of the glucose level in the blood plasma has been achieved in vitro and in vivo by administration of vanadium and zinc in form of inorganic salts. It has been shown that elements are poorly absorbed in their inorganic forms and required high doses which have been associated with undesirable side effects. Many researchers, therefore, have focused on metal complexes that were prepared from VOSO4 or ZnSO4 and low-molecular-weight bidentate ligands. Seven kinds of 1-hydroxy-4,6-disubstituted and 1-hydroxy-4,5,6-trisubstituted-2(1H)-pyrimidinones were synthesized by reaction of N-benzyloxyurea and β-diketones and subsequent removal of the protecting group. Six kinds of 1-hydroxy-4-(substituted)amino-2(1H)-pyrimidinones were synthesized by the substitution reaction of 1-benzyloxy-4-(1′,2′,4′-triazol-1′-yl)-2(1H)-pyrimidinone with various alkyl amines or amino acids. Treatment with VOSO4 and ZnSO4 or Zn(OAc)2 afforded vanadyl(IV) and zinc(II) complexes which were characterized by means of 1H NMR, IR, EPR, and UV–vis spectroscopies, and combustion analysis. The in vitro insulin–mimetic activity of these complexes was evaluated from 50% inhibitory concentrations (IC50) on free fatty acid (FFA) release from isolated rat adipocytes treated with epinephrine. Vanadyl complexes of 4,6-disubstituted-2(1H)-pyrimidinones showed higher insulin–mimetic activities than those of 4,5,6-trisubstituted ones. On the other hand, Zn(II) complexes showed lower insulin–mimetic activities than VOSO4 and ZnSO4 as positive controls. It was found that the balance of the hydrophilicity and/or hydrophobicity is important for higher insulin–mimetic activity. The in vivo insulin–mimetic activity was evaluated with streptozotocin (STZ)-induced diabetic rats. Blood glucose levels were lowered from hyperglycemic to normal levels after the treatment with bis(1,2-dihydro-4,6-dimethyl-2-oxo-1-pyrimidinolato)oxovanadium(IV) by daily intraperitoneal injections. The improvement in glucose tolerance was also confirmed by an oral glucose tolerance test. [Copyright &y& Elsevier]

The thin-layer chromatographic behavior of alkaline earth metal ions (Mg2+, Ca2+, Sr2+, and Ba2+) in aqueous sodium perchlorate solutions on silica gel thin-layers impregnated with sodium hydroxide has been surveyed as a function of salt concentration. At salt concentrations above 2moll-1, the selectivity of the metals increased with a decrease in the crystal ionic radii; with further increases in salt concentration, the selectivity differences among the metals expanded remarkably. In the present systems, it was supposed that the cation exchange, the surface complexation, and the salting-out effect participate simultaneously in the adsorption of the metals on silica gel. Typical chromatograms for the mutual separation of the alkaline earth metals are presented. [Copyright &y& Elsevier]

The high affinity and specificity of nucleic acid base complementarity has been proven to be a powerful method for constructing specific molecular assemblies. On the other hand, recent structural studies of RNA have revealed the wide range of tertiary interactions utilized in RNA folding, which may potentially be used as tools for the design of specific macromolecular assemblies. Here, RNA building blocks containing two hairpin loops, based on the dimerization initiation site (DIS) of HIV RNA, connected by a short linker were used to construct large RNA assemblies through hairpin loop-loop (“kissing”) interactions. We show that specific linear and circular assemblies can be constructed in a magnesium-dependent manner using several non-self-complementary loop-loop interactions designed in this study. These results show that the use of RNA tertiary interactions may broaden the reportoire of nucleic acid-based nanostructures. [Copyright &y& Elsevier]

The growth and spreading of fibroblast, L929 cells, on various polymeric films prepared by the Langmuir-Blodgett (LB) and casting methods were investigated. L929 cells, which were cultivated on collagen and synthetic polymeric films prepared by the LB method, adhered and spread much more than those on synthetic films prepared by the casting method. This is explained by the fact that cell growth and cell spreading are suitable for L929 cells on the films having serum proteins that contain a high α-helix content, because LB films adsorbed those serum proteins estimated from the circular dichroism measurements of the films immersed in cell culture medium. An exponential relationship was observed from the plot of the cell density vs root mean square of roughness of the films, which is estimated by atomic force microscopy, whereas a linear relationship was observed from the plot of the spreading ratio vs the root mean square of roughness. It is suggested that the correlation between the cell growth or spreading ratio and surface roughness of the films where L929 cells were cultivated is considered to be more important than the correlation between the cell growth or spreading ratio and the contact angle of the films. [ABSTRACT FROM AUTHOR]