Melatonin-Induced Postconditioning Suppresses NMDA Receptor through Opening of the Mitochondrial Permeability Transition Pore via Melatonin Receptor in Mouse Neurons

Mitochondrial membrane potential regulation through the mitochondrial permeability transition pore (mPTP) is reportedly involved in the ischemic postconditioning (PostC) phenomenon. Melatonin is an endogenous hormone that regulates circadian rhythms. Its neuroprotective effects via mitochondrial melatonin receptors (MTs) have recently attracted attention. However, details of the neuroprotective mechanisms associated with PostC have not been clarified. Using hippocampal CA1 pyramidal cells from C57BL mice, we studied the involvement of MTs and the mPTP in melatonin-induced PostC mechanisms similar to those of ischemic PostC. We measured changes in spontaneous excitatory postsynaptic currents (sEPSCs), intracellular calcium concentration, mitochondrial membrane potential, and N-methyl-D-aspartate receptor (NMDAR) currents after ischemic challenge, using the whole-cell patch-clamp technique. Melatonin significantly suppressed increases in sEPSCs and intracellular calcium concentrations. The NMDAR currents were significantly suppressed by melatonin and the MT agonist, ramelteon. However, this suppressive effect was abolished by the mPTP inhibitor, cyclosporine A, and the MT antagonist, luzindole. Furthermore, both melatonin and ramelteon potentiated depolarization of mitochondrial membrane potentials, and luzindole suppressed depolarization of mitochondrial membrane potentials. This study suggests that melatonin-induced PostC via MTs suppressed the NMDAR that was induced by partial depolarization of mitochondrial membrane potential by opening the mPTP, reducing excessive release of glutamate and inducing neuroprotection against ischemia-reperfusion injury.     

Negative staining--carbon replica method, a new procedure to reveal ultrastructure of freeze-fractured membrane.

The present study was undertaken to report a new method which reveals the ultrastructure of freeze-fractured membrane faces. The method consists of initial replication of fractured surfaces of frozen specimens with evaporated carbon and treatment of the replicas with a negative staining solution after dissolution of specimens. This negative staining-carbon replica method reveals clear distribution, ultrastructures and precise size of intramembrane particles on the fractured membrane faces by means of density difference of negative staining solution which is deposited on the surfaces of carbon replicas.     

Relationship between local structure and stability in hen egg white lysozyme mutant with alanine substituted for glycine

We prepared five mutant lysozymes in which glycines whose dihedralangles are located in the region of the left-handed helix, Gly49,Gly67, Gly71, Gly102 and Gly117, were mutated to an alanineresidue. From analyses of their thermal stabilities using differentialscanning calorimetry, most of them were more destabilized thanthe native lysozyme, except for the G102A mutant, which hasa stability similar to that of the native lysozyme at pH 2.7.As for the destabilized mutant lysozymes, their X-ray crystallographicanalyses showed that their global structures did not changebut that the local structures changed slightly. By examiningthe dihedral angles at the mutation sites based on X-ray crystallographicresults, it was found that the dihedral angles at these mutationsites tended to adopt favorable values in a Ramachandran plotand that the extent and direction of their shifts from the originalvalue had similar tendencies. Therefore, the change in dihedralangles may be the cause of the slight local structural changesaround the mutation site. On the other hand, regarding the mutationof G102A, the global structure was almost identical with thatof the native structure but the local structure was drasticallychanged. Therefore, it was suggested that the drastic localconformational change might be effective in releasing the unfavorableinteraction of the native state at the mutation site.     

Keratanase II-catalyzed synthesis of keratan sulfate oligomers by using sugar oxazolines as transition-state analogue substrate monomers: a novel insight into the enzymatic catalysis mechanism

Keratan sulfate (KS) oligomers with well-defined structures were synthesized by keratanase II (KSase II)-catalyzed transglycosylation. N-Acetyllactosamine [Galbeta(1-->4)GlcNAc; LacNAc] oxazoline derivatives with sulfate groups at the C-6 (1 a) and both the C-6 and the C-6' (1 b) were prepared as transition-state analogue substrate monomers for KSase II. Monomer 1 a was effectively oligomerized by the enzyme under weak alkaline conditions, to give alternating 6-sulfated KS oligomers (2 a) in good yields, and with total control of regioselectivity and stereochemistry. KSase II also recognized 1 b, which provided fully 6-sulfated KS oligomers (2 b) in good yields under similar conditions. Nonsulfated LacNAc oxazoline was difficult to oligomerize enzymatically. These results imply that the catalysis mechanism of KSase II involves a sugar oxazolinium ion that requires the 6-sulfate group in the GlcNAc residue not only in hydrolysis of KS chains, but also in oligomerization of oxazoline monomers. This is the first report of KSase II-catalyzed transglycosylation to form beta(1-->3)-glycosidic bond through a substrate-assisted mechanism.     

Change of orientation by thermal contraction of polyacrylonitrile fiber

Polyacrylonitrile polymer powder was dissolved in 70% nitric acid and spun into isotropic filament through a glass nozzle of 0.5 mm. diameter in a coagulating bath of 30% nitric acid. Stretching was carried out in two stages: the first stretching was done in water at 20°C. followed by drying, and the second stretching was done in a boiling saturated solution of ammonium sulfate. The total stretching ratio was 23. These filaments were shrunk freely in water at 70–180°C. The change in orientation factors was traced by x-ray, infrared dichroism, visible dichroism, and sonic modulus methods. The relation between the reciprocal absolute temperature of thermal contraction and the logarithm of fiber length is a straight line which has two inflection points at 93 and 175°C. The orientation factors by x-ray and infrared dichroism remain unchanged up to 175°C. On the contrary, the orientation factors by visible dichroism and sonic modulus drop suddenly at about 90°C. This indicates the occurrence of relaxation of the amorphous chain at the glass transition temperature and shows the polymer is not perfect single-phase material. Orientation of crystalline and amorphous phases is stable from 100 to 170°C. in spite of considerable thermal contraction. The stability of orientation can be explained by the growth of a folded structure in the polymer.     

Poly(4-diphenylaminostyrene) with a well-defined polymer chain structure: Controllable optical and electrical properties

The effect of polymer chain structure on the optical and electrical properties are reported for poly(4-diphenylaminostyrene) (PDAS), which was prepared by the living anionic polymerization of 4-diphenylaminostyrene (DAS) with the benzyllithium (BzLi)/N,N,N′,N′-tetramethylethylenediamine (TMEDA) system. The optical properties of PDAS are strongly affected by the stereoregularity of the PDAS polymer chain; intramolecular excimer-forming fluorescence was observed from PDAS with a syndiotactic-rich configuration. The highest occupied molecular orbital (HOMO) and lowest unoccupied molecular orbital (LUMO) energy levels of PDAS were approximately −5.4 and −2.0 eV, respectively, regardless of the polymer chain structure. The hole and electron drift mobilities for PDAS were in the order of 10⁻⁴ to 10⁻⁵ (cm²/V s) and 10⁻⁵ (cm²/V s), respectively, with negative slopes. The distance of each triphenylamino (TPA) group in the polymer chain was a major factor influencing the drift mobility of PDAS. The current-voltage (I-V) characteristics of PDAS were controllable according to the polymer chain structure of PDAS.     

Reaction of Syringylglycerol-β-Syringyl Ether Type of Lignin Model Compounds in Alkaline Medium

Reactions of three kind of syringylglycerol- β-syringyl ether type model compounds under alkaline medium were investigated. Sinapyl alcohol and β-hydroxypropiosyringone were formed as phenyl propanoid moieties from syringylglycerol- β-(methyl-syringyl) ether 1 by the β-aryl ether cleavage under soda treatment, while only sinapyl alcohol was formed from syringylglycerol- β-syringyl ether 2. The formation of both two degradation products are quite interesting because there is no nucleophilic additives in soda liquor. A possible reaction mechanisms for the β-aryl cleavage of syringylglycerol-β-syringyl ether type is homolytical cleavage via quinone methide.     

The spatial distribution of Zn during galvanic corrosion of a Zn/steel couple

The spatial distribution of Zn²⁺ during galvanic corrosion of a model Zn/steel couple in 0.01 M NaCl was investigated using a scanning zinc disk electrode. The couple had a coplanar arrangement of a steel substrate with an electroplated zinc layer at the center. During galvanic corrosion, the marked changes in the Zn²⁺ concentration were confined to a thin solution layer ca. 1.0 mm thick above the couple surface. In this thin solution layer above the zinc layer, a higher concentration region of Zn²⁺ in the range of 5-18 mM extended around the zinc layer in the solution during galvanic corrosion. Conversely, above the steel surface distant from the zinc layer, the surface concentration of Zn²⁺ was almost zero during galvanic corrosion. On this surface, the precipitation of zinc corrosion products due to the hydrolysis reaction of Zn²⁺ was observed. The distribution of the Zn²⁺ concentration supported that Zn²⁺ acted as a buffer that suppressed the increased pH due to the cathodic reaction on the steel surface near the zinc layer and almost no corrosion products formed there. The spatial distribution of Zn²⁺ is discussed in relation to the distributions of potential and pH and the surface morphology of the galvanic couple.     

Ultra-high pressure sterilization of powdery food stuff—a new application of a roller compactor

A new non-heating technique was developed for the sterilization of food stuff. Applying a roller compactor, ultra-high pressure sterilization has experimentally demonstrated its ability to sterilize dry powders, such as corn flour and Chinese herbs, with little quality deterioration. Also, the degree of food sterilization was found correlated well with the linear press forces between the rollers, roller gaps, and number of compaction passes. In comparison to the conventional high pressure sterilization technique, the new dry continuous processing method has the advantage of lower investment cost and is more versatile for sterilizing various food powders.