Chromatoid bodies: aggresome-like characteristics and degradation sites for organelles of spermiogenic cells.

We investigated the localization of several markers for lysosomes and aggresomes in the chromatoid bodies (CBs) by immunoelectron microscopy. We found so-called aggresomal markers such as Hsp70 and ubiquitin in the core of the CBs and vimentin and proteasome subunit around the CBs. Ubiquitin-conjugating enzyme (E2) was also found in the CBs. In tubulovesicular structures surrounding the CBs, lysosomal markers were detected but an endoplasmic reticulum retention signal (KDEL) was not. Moreover, proteins located in each subcellular compartment, including the cytosol, mitochondria, and nucleus, were detected in the CBs. Signals for cytochrome oxidase I (COXI) coded on mitochondrial DNA were also found in the CBs. Quantitative analysis of labeling density showed that all proteins examined were concentrated in the CBs to some extent. These results show that the CBs have some aggresomal features, suggesting that they are not a synthetic site as proposed previously but a degradation site where unnecessary DNA, RNA, and proteins are digested.     

Heterogeneity in Expression of Lipopolysaccharide and Major Outer-Membrane Proteins by Strains of Escherichia coli O157 with Different H-Serotypyes

A total of 11 strains of Escherichia coli (E. coli) belonging to serogroup O157 were examined for the expression of long-chain lipopolysaccharide (LPS) and major outer-membrane proteins (OMPs) by means of SDS-PAGE. The strains belonged to either one of four different flagellar (H) types or did not express flagella. Four of the eleven strains carried genes encoding Shiga-like toxins (SLTs). All the strains exhibited one of four LPS profiles, designated A, B, C or D. Electron microscopic analysis with the freeze-substitution technique demonstrated the differences in the cell surface structures of strains with each LPS profile. Strains with LPS profile A, B or C had layers of thin fibers 10, 20 and 20 nm long, respectively, on the outer membrane but strains with LPS profile D had no such structure. An analysis of the OMPs showed that all the strains had one of four OMP profiles, designated I, II, III or IV. Both LPS and OMP profiles were dependent on H-serotypes, and the combination pattern of LPS and OMP profiles of the strains was unique for each H-serotype. These data support the existence of heterogeneous groups of O157 strains.     

Discovery of SMP-304, a novel benzylpiperidine derivative with serotonin transporter inhibitory activity and 5-HT1A weak partial agonistic activity showing the antidepressant-like effect

We report the discovery of a novel benzylpiperidine derivative with serotonin transporter (SERT) inhibitory activity and 5-HT_1A receptor weak partial agonistic activity showing the antidepressant-like effect. The 3-methoxyphenyl group and the phenethyl group of compound 1, which has weak SERT binding activity, but potent 5-HT_1A binding activity, were optimized, leading to compound 35 with potent and balanced dual SERT and 5-HT_1A binding activity, but also potent CYP2D6 inhibitory activity. Replacement of the methoxy group in the left part of compound 35 with a larger alkoxy group, such as ethoxy, isopropoxy or methoxy-ethoxy group ameliorated CYP2D6 inhibition, giving SMP-304 as a candidate. SMP-304 with serotonin uptake inhibitory activity and 5-HT_1A weak partial agonistic activity, which could work as a 5-HT_1A antagonist, displayed faster onset of antidepressant-like effect than a representative SSRI paroxetine in an animal model.     

Establishment and yearly/seasonal occurrence of the exotic coccidophagous ladybird <Emphasis Type="Italic">Cryptolaemus montrouzieri</Emphasis> (Coleoptera: Coccinellidae) in citrus groves in Shizuoka City,central Japan: a 5-year survey on adult numbers

Cryptolaemus montrouzieri Mulsant (Coleoptera: Coccinellidae) is a ladybird native to Australia, preying on mealybugs and soft scales, and has been utilized worldwide as a biological control agent. It has long been recognized that C. montrouzieri that was introduced into the main island of Japan had failed to become established. The present study monitored yearly and seasonal occurrence of C. montrouzieri adults in citrus groves at Shizuoka Prefectural Fruit Tree Research Center in Shizuoka City, central Japan in 2008–2012 by using sticky traps and beating citrus trees. Adults of C. montrouzieri were continuously captured for 5 and 4 years in a pesticide-free citrus grove and a neighboring reduced-pesticide grove, respectively. Larvae of C. montrouzieri were observed consuming a cottony scale, Pulvinaria aurantii Cockerell, on citrus trees. These results provide unequivocal evidence for the ladybird’s establishment in central Japan. The number of trapped ladybird adults exhibited four peaks a year: in mid-April, early to late June, mid-August, and late September to early October. Adult numbers in each grove varied largely across years, showing a great increase followed by a rapid decline during a period of 4 years. Factors affecting the seasonal/yearly occurrence of C. montrouzieri adults in citrus groves are discussed.     

Reaction equilibrium for lipase-catalyzed condensation in organic solvent systems

Lipase-catalyzed condensation in an organic solvent is useful for the syntheses of esters. To reasonably design and optimize the reaction conditions, knowledge of the reaction equilibrium is required. The interaction of water with other reactants and the quantitative predictions for adsorption of water by a desiccant are discussed. The solvent effects on the reaction equilibrium are also elucidated in mixtures of nitrile and tert-alcohol.     

Application of heat-induced antigen retrieval to aldehyde-fixed fresh frozen sections.

We applied the heat-induced antigen retrieval (HIAR) to aldehyde-fixed fresh frozen sections based on a new approach (i.e., a rapid and complete immobilization of antigen followed by heating). Frozen sections were fixed with 10% formalin in 0.1 M cacodylate buffer (pH 7.4) containing 25 mM CaCl(2) for 30 min, or with 0.5% glutaraldehyde in 0.1 M phosphate buffer (pH 7.4) for 1 min at room temperature, and then autoclaved in 20 mM Tris-HCl buffer (pH 9.0) for 10 min at 120 C. Both fixatives yielded good tissue structure after autoclaving. In the sections fixed with formalin containing CaCl(2), 20 of 22 antigens located in the nucleus, cytoplasm, membranes, and extracellular matrix greatly recovered their antigenicity after autoclaving; only two antigens exhibited stronger immunoreaction in acetone-fixed fresh frozen sections than these sections. Heating also retrieved the immunoreactivity of at least 14 antigens in the sections fixed with glutaraldehyde. We used the similar procedures to localize ligand-free estrogen receptor alpha (ERalpha) and glucocorticoid receptors (GR). Mouse uterine cells exhibited almost the same nuclear ERalpha immunostaining regardless of the hormonal status in glutaraldehyde-fixed fresh frozen sections and unliganded GR was localized mainly in the nucleus of mouse hepatocytes in fresh frozen sections fixed with 20% formalin containing 50 or 75 mM CaCl(2) at 40 C, after autoclaving. These results demonstrate that HIAR is useful for the immunohistochemistry of many antigens in aldehyde-fixed fresh frozen sections.     

Rod and cone photoreceptors: molecular basis of the difference in their physiology

Vertebrate retinal photoreceptors consist of two types of cells, the rods and cones. Rods are highly light-sensitive but their flash response time course is slow, so that they can detect a single photon in the dark but are not good at detecting an object moving quickly. Cones are less light-sensitive and their flash response time course is fast, so that cones mediate daylight vision and are more suitable to detect a moving object than rods. The phototransduction mechanism was virtually known by the mid 80s, and detailed mechanisms of the generation of a light response are now understood in a highly quantitative manner at the molecular level. However, most of these studies were performed in rods, but not in cones. Therefore, the mechanisms of low light-sensitivity or fast flash response time course in cones have not been known. The major reason for this slow progress in the study of cone phototransduction was due to the inability of getting a large quantity of purified cones to study them biochemically. We succeeded in its purification using carp retina, and have shown that each step responsible for generation of a light response is less effective in cones and that the reactions responsible for termination of a light response are faster in cones. Based on these findings, we speculated a possible mechanism of evolution of rods that diverged from cones.     

Extraction of defatted rice bran by subcritical water treatment

Defatted rice bran was treated with subcritical water in the temperature range of 180–280 °C for 5 min using 117 mL and 9 mL vessels to produce the extracts. The total sugar and protein contents and radical scavenging activity of the extracts were then estimated for both vessels. The total sugar concentration of ca. 0.3 g/L-extract was the highest for the extracts at 200 °C, and it significantly decreased at the higher temperatures. The protein concentration and radical scavenging activity were higher at the higher temperatures. Extraction was also done at 200 °C and 260 °C for various times using the small vessel. The total sugar concentration decreased with the increasing extraction time, while the protein concentration and radical scavenging activity only slightly depended on the extraction time. The extracts at 200 °C or lower temperatures using the large vessel possessed the emulsifying and emulsion-stabilizing activities. The HPLC analysis of the extract at 260 °C for 5 min using the small vessel indicated that it contained both hydrophilic and hydrophobic substances. The hydrophilic fraction of the extract mainly contained low-molecular-mass substances.