One-Pot Tandem Coupling Method for the Short-Step Formal Synthesis of Riccardin C.

One-pot reactions reduce reagent amounts and circumvent process treatments, such as work-up and purifications in multi-step reactions. In this study, we achieved the formal total synthesis of riccardin C through a one-pot reaction by simultaneously linking four units through two Sonogashira coupling reactions and one Suzuki coupling reaction, followed by reduction and deprotection. Thus, this one-pot method comprised five steps and did not require the purification of intermediate reaction mixtures, which saves resources, such as reagents and solvents, and expedites the work process.

Chlamydial Infection-Dependent Synthesis of Sphingomyelin as a Novel Anti-Chlamydial Target of Ceramide Mimetic Compounds.

The obligate intracellular bacterium Chlamydia trachomatis is the major causative agent of bacterial sexually transmitted diseases worldwide. In infected cells, the ceramide transport protein (CERT) is recruited to inclusions, where C. trachomatis replicates using host-synthesized ceramide. The ceramide is converted to sphingomyelin (SM) by a chlamydial infection-dependent SM synthesis (cidSM-synthesis) pathway, which occurs even in the absence of the SM synthases (SMS)-1 and -2 of host cells. The ceramide mimetic compound (1R,3S)-HPA-12 and the nonmimetic compound E16A, both of which are potent inhibitors of CERT, repressed the proliferation of C. trachomatis in HeLa cells. Unexpectedly, (1R,3R)-HPA-12, a ceramide mimetic compound that lacks CERT inhibitory activity, also exhibited potent anti-chlamydial activity. Using endogenous SMS-knockout mutant HeLa cells, we revealed that (1R,3R)-HPA-12 mildly inhibited cidSM-synthesis. In addition, LC-MS analysis revealed that (1R,3R)-HPA-12 is converted to a phosphocholine-conjugated metabolite in an infection-dependent manner. Imaging analysis with a fluorescent analog of ceramide suggested that cidSM-synthesis occurs in the bacterial bodies and/or inclusions. Collectively, these results suggested that (1R,3R)-HPA-12 exerts its anti-chlamydia activity not only as an inhibitor of cidSM-synthesis, but also via putative toxic effects of its phosphocholine adduct, which is most likely produced by the cidSM-synthesis route.

Retrieval-induced forgetting in rats.

"Retrieval-induced forgetting" in rats was evaluated using a modified spontaneous object recognition test. The test consisted of a sample phase, retrieval or interference phase, and a test phase with 60-min delay period inserted between the phases. Rats were randomly assigned to one of three groups (control, retrieval and interference) and allowed to explore the field in which two different objects (A, B) were placed in the sample phase. In the retrieval phase, two identical objects (B, B), which were the same as one of the objects presented in the sample phase, were placed again. In the interference phase, two identical objects (C, C), which were novel for animals, were placed. In the test phase, two different objects (A, D), one of which was identical to that presented in sample phase (familiar object) and the other was novel, were placed and the time spent exploring each object was analyzed. While the exploration of the novel object was significantly longer than that of the familiar object in rats subjected to the interference phase, rats subjected to the retrieval phase could not discriminate between the familiar and the novel objects at the test phase. These results demonstrate the "retrieval-induced forgetting" phenomenon in a spontaneous object recognition test in rats.

Relationships between drinking habits, psychological resilience, and salivary cortisol responses on the Trier Social Stress Test-Online among Japanese people.

BACKGROUND: This study aimed to determine whether individual differences in resilience interacted with those in alcohol consumption habits in situations involving exposure to psychosocial stressors (Trier Social Stress Test-Online; TSST-OL). Additionally, we investigated whether individuals exhibiting resilience in their psychological scale scores showed biological responses that could be interpreted as resilience in stressful situations, such as the TSST-OL. We hypothesized that there would be no association between drinking habits and stress responses in the high-resilience group. Furthermore, high drinking habits would be associated with high stress responses in the low-resilience group. METHODS: We recruited 22 and 20 individuals from the high and low-resilience groups, respectively, from among those who completed the online survey comprising the Alcohol Use Disorders Identification Test (AUDIT) and resilience scales; we excluded individuals with AUDIT scores of 15 or higher, and divided them by the median total resilience scale score. During the TSST-OL, self-rated stress measurement and saliva sample collection were performed seven times. Frozen samples were collected at the Tokyo site, and salivary hormonal (cortisol and dehydroepiandrosterone) levels were measured after transport in frozen state. Finally, 36 participants were included in the analysis of self-rated stress and cortisol levels. RESULTS: We observed the typical subjective stress responses to the TSST-OL. People with higher psychological scale scores for resilience traits showed significantly higher salivary cortisol levels than those with lower scores. Due to deficiencies in the survey and experimental design, the classification criteria were changed and an exploratory analysis was performed to investigate the interaction of individual differences in resilience and drinking habits. In contrast to our expectation, those with low resilience scores showed stress responses, regardless of their drinking habits. Furthermore, those with high resilience and drinking habits showed a specific insensitivity to salivary cortisol levels. Their self-rated stress scores were similar to those of other groups. CONCLUSIONS: Our study showed the applicability of the TSST-OL in the Japanese population, the individual relationship between psychological resilience measures and biological stress responses, and a specific insensitivity in the salivary cortisol response as a result of individual differences in high resilience and drinking habits.

Bismuth catalysts in aqueous media.

Several bismuth-catalyzed synthetic reactions, which proceed well in aqueous media, are discussed. Due to increasing demand of water as a solvent in organic synthesis, catalysts that can be used in aqueous media are becoming more and more important. Although bismuth Lewis acids are not very stable in water, it has been revealed that they can be stabilized by basic ligands. Chiral amine and related basic ligands combined with bismuth Lewis acids are particularly useful in asymmetric catalysis in aqueous media. On the other hand, bismuth hydroxide is stable and works as an efficient catalyst for carbon-carbon bond-forming reactions in water.

Chiral-Sc catalyzed asymmetric Michael addition/protonation of thiols with enones in water.

Asymmetric Michael reactions and enantioselective protonations between enones and thiols were catalyzed by a Sc(OTf)(3)-chiral 2,2'-bipyridine complex in water. The remarkable governing of the enantioselectivity for simple introduction of protons despite their abnormally high mobility in water may provide us with new synthetic opportunities as well as significant chemical advances.

Practical method for hydroxyl-group protection using strontium metal and readily available silyl chlorides.

We have found that the etherification of silyl-protected secondary alcohols proceeds smoothly in the presence of strontium metal using silyl chloride instead of the expensive, yet more reactive, and commonly used silyl triflate. The reaction occurred almost completely with various alcohols.

Chiral Sc-catalyzed asymmetric Michael reactions of thiols with enones in water.

Asymmetric Michael reactions of thiols with enones were catalyzed by a Sc(OTf)(3)-chiral bipyridine complex at room temperature in water without using any organic solvents, to afford the desired sulfides in high yields with high enantioselectivities.

A gold-immobilized microchannel flow reactor for oxidation of alcohols with molecular oxygen.

Golden capillaries: A gold-immobilized capillary column reactor allows oxidation of alcohols to carbonyl compounds using molecular oxygen. These capillary columns (see picture) can be used for at least four days without loss of activity.

Catalytic use of indium(0) for carbon-carbon bond transformations in water: general catalytic allylations of ketones with allylboronates.

We have discovered the unprecedented catalytic use of In(0) for catalytic C-C bond transformations. Remarkably, these general catalytic allylations of ketones proceeded smoothly in water as a sole solvent under mild conditions, and water proved to be essential for these reactions. Both the displayed substrate scope and the functional group tolerance were excellent. Importantly, the In metal catalyst could be easily recovered and reused without loss of catalytic activity. Moreover, when an alpha-substituted allylboronate was used, an unusual constitutional selectivity was observed providing exclusively the formal alpha-adduct. Additionally, the resulting tertiary homoallylic alcohols were obtained with exceptionally high diastereoselectivities. The applicability of this concept to asymmetric catalysis in water by using In(0) combined with a chiral bis(oxazoline) ligand was demonstrated as well.

Development of microchannel reactors using polysilane-supported palladium catalytic systems in capillaries.

A new method of immobilizing Pd catalysts on the channel wall of a capillary by using polysilane with metal oxide has been developed, and applied to hydrogenation reactions.

[Mechanism of micelle-vesicle transformation and control of vesicular sizes and properties].

The mechanism of vesicle-to-micelle or micelle-to-vesicle transition was studied in order to control sizes and fluidities of vesicles during periods of preparation. Dependence of particle sizes measured by quasi-elastic light scattering, turbidities, fluidity parameters monitored by ESR spectroscopy, and morphological changes of mixed aggregates of egg yolk phosphatidylcholine (EPC) and a detergent (octylglucoside (OG) or sodium cholate (Na-chol)) on detergent concentration provided a model of vesicle destruction. It possessed three phase transition points, and proceeded in a stepwise fashion: vesicles, small particles containing large amounts of detergents (SUV(*)), intermediate structures, and mixed micelles. Vesicle formation on removal of detergents from micelles proceeded oppositely. Micelle-vesicle transition mechanism was common to all detergents examined. The feature of the mechanism was the presence of SUV(*). Next, SUV(*) was prepared by adding appropriate amount of a detergent to small unilamellar vesicles obtained by sonication. Time-dependent size growth of the SUV(*) was remarkable in the case of OG-containing SUV(*), but was insignificant in the case of Na-chol-containing SUV(*), suggesting the size determining step to be the stage of the SUV(*). The tendency to produce large or small vesicles from micelles was related to the absence or presence, respectively, of a net charge in the detergent molecule. The fluidities of EPC micelles containing small amounts of a detergent possessing a steroidal structure (e.g., Na-chol or CHAPS) were significantly smaller than the corresponding values of a detergent without a steroidal structure (e.g., OG), suggesting a method of control of orderliness of hydrocarbon chains in EPC vesicles.

The relationship between fear extinction and resilience to drug-dependence in rats.

Some individuals recover quickly from stressful events, while others do not; this suggests that there are individual differences in stress resilience. Previous studies in rats have shown that an animal's ability to extinguish conditioned fear can predict their stress resilience. In the present study, we investigated whether rats who are resilient or vulnerable to fear conditioning would consistently show resilient or vulnerable behaviors in other behavioral tests. Male Wistar-Imamichi rats were subjected to a series of behavioral tests, including open field, conditioned place-preference (CPP), forced swim and auditory fear conditioning. The subjects were assigned to one of two different groups: resilient or vulnerable, based on their level of freezing during fear extinction, and their other behavioral phenotypes were then compared. Rats in the resilient group showed faster extinction in the methamphetamine-induced CPP test than rats in the vulnerable group. The results suggest that resilience to stressful events is associated with drug-dependence and that individuals' abilities to recover from stressful situations and drug-dependence may share a common mechanism.

Selective Hydrogenation of Nitriles to Primary Amines Catalyzed by a Polysilane/SiO2-Supported Palladium Catalyst under Continuous-Flow Conditions.

Hydrogenation of nitriles to primary amines with heterogeneous catalysts under liquid-phase continuous-flow conditions is described. Newly developed polysilane/SiO2-supported Pd was found to be an effective catalyst and various nitriles were converted into primary amine salts in almost quantitative yields under mild reaction conditions. Interestingly, a complex mixture was obtained under batch conditions. Lifetime experiments showed that this catalyst remained active for more than 300 h (TON≥10 000) without loss of selectivity and no metal leaching from the catalyst occurred. By using this continuous-flow hydrogenation, synthesis of venlafaxine, an antidepressant drug, has been accomplished.

Supercooled micro flows and application for asymmetric synthesis.

Supercooled micro flow in microchannels was demonstrated. In order to clarify fundamental properties of the supercooling state in microchannels, freezing temperature was measured in the microchannels having widths ranging from 70 microm to 300 microm. The freezing temperature decreased with decreasing width of the microchannel when the microchannel wall was chemically modified with octadecylsilane group. The lowest freezing temperature was observed as -28 degrees C for water in the 70 microm wide microchannel. By contrast, the freezing temperature was -15 degrees C and did not depend on width when a microchannel with a bare glass surface was used. Next, the freezing point was measured with flow rates ranging from 0.1 to 2.0 microl min(-1) and no dependence on flow rate was observed. Then, the supercooled micro flow was applied to an asymmetric reaction in micro two-phase flow of aqueous and CH2Cl2 phases. As expected from thermodynamic prediction, enantiomeric selectivity increased in the supercooled state of water.

Delivery of condensed DNA by liposomal non-viral gene delivery system into nucleus of dendritic cells.

In this study, we developed novel double-membranous non-viral gene delivery system modified with SV-40 T antigen-derived nuclear localization signal (NLS-DMEND) for delivery of luciferase plasmid DNA to nucleus of non-dividing mouse bone marrow-derived dendritic cells (BMDC). Intracellular trafficking and gene expression of NLS-DMEND in the BMDC were evaluated. Condensed DNA was observed in the nucleus by confocal laser scanning microscopy, and the NLS-DMEND induced significant luciferase activity in the BMDC. It was suggested that the condensed DNA particle transferred into nucleus via energy dependent manner, since the nuclear transfer was inhibited by metabolic inhibitors. In conclusion, condensed plasmid DNA was delivered into the nucleus of non-dividing BMDC by NLS-DMEND.

Construction of a multifunctional envelope-type nano device by a SUV*-fusion method.

A novel assembly method "SUV*-fusion method" was developed for the construction of a small and homogenous multifunctional envelope-type nano device (MEND) by utilizing a detergent-rich small unilamellar vesicle (SUV*). The method consists of three steps: (1) DNA condensation with a polycation, (2) electrostatic interaction of the SUV* with the DNA/polycation complex (DPC) and (3) lipid coating of DPC by SUV* fusion via removal of the detergent. We confirmed the construction of the MEND by sucrose density gradient centrifugation, and isolated the MEND only from the boundary between 25% and 40% sucrose. The isolated MEND had a small diameter (155 nm), was negatively charged (-24 mV), and encapsulated 30% of the total DNA. The MEND was formed by only SUV*, not by a lipid/detergent micelle. This confirms that a small and homogenous MEND can be constructed by the SUV*-fusion method. Furthermore, we confirmed that a transferrin-modified MEND could deliver a gene into a cell through receptor-mediated endocytosis. Consequently, we report on the successful construction of a small and homogenous MEND by a novel SUV*-fusion method.

Air-stable, storable, and highly selective chiral Lewis acid catalyst.

An air-stable, storable, and highly selective chiral Lewis acid catalyst for asymmetric Mannich-type reactions has been developed. The catalyst can be stored for more than three months in air at room temperature without loss of activity. Moreover, it has also been demonstrated that the catalyst can be recovered and reused. [structure: see text]