Synthesis of biologically active cyclic peptides

1. The extent of racemization and the coupling yield in peptide synthesis were studied under high dilution conditions. The azide method yielded the best results. 2. Five linear penta-peptide precursors related to gramicidin S were subjected to cyclization in order to study how the difference in the sequence influences the yield and the ratio of cyclic dimer to monomer. The azide with the sequence of -L -Pro-L -Val-L -Orn(Z)-L -Leu-D -Phe- afforded diZ-gramicidin S in a high yield of 63%. 3. Alternaria mali toxin III, a cyclotetradepsipeptide phytotoxin, was synthesized. The activated linear tetradepsipeptide containing a D -Dap(Z) (N³-Z-D -2,3-diaminopropionic acid) residue at the N-terminus afforded the cyclic precursor (53%). The Dap residue in the precursor was converted into a ΔAla residue by Hofmann degradation to give the desired product.     

Pyrazolo pyrimidine-type inhibitors of SRC family tyrosine kinases promote ovarian steroid-induced differentiation of human endometrial stromal cells in vitro

Reversible protein tyrosine phosphorylation, coordinately controlled by protein tyrosine kinases and phosphatases, is a critical element in signal transduction pathways regulating a wide variety of biological processes, including cell growth, differentiation, and tumorigenesis. We have previously reported that c-Src belonging to the Src family tyrosine kinase (SFK) becomes dephosphorylated at tyrosine 530 (Y530) and thereby activated during progestin-induced differentiation of human endometrial stromal cells (i.e., decidualization). In this study, to elucidate the role of decidual c-Src activation, we examined whether 4-amino-5-(4-methylphenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine (PP1) and 4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine (PP2), both potent and selective SFK inhibitors, affected the ovarian steroid-induced decidualization in vitro. Unexpectedly, PP1 paradoxically increased the kinase activity of decidual c-Src together with dephosphorylation of Y530 in the presence of ovarian steroids. Concomitantly, PP1 enhanced morphological and functional decidualization, as determined by induction of decidualization markers, such as insulin-like growth factor binding protein-1 and prolactin. PP2 also advanced decidualization along with up-regulation of the active form of c-Src whose Y-530 was dephosphorylated. In contrast to PP1 and PP2, herbimycin A, a tyrosine kinase inhibitor with less specificity for SFKs, showed little enhancing effect on the expression of both IGFBP-1 and active c-Src. These results suggest that SFKs, including c-Src, may play a significant role in stromal cell differentiation, providing a clue for a possible therapeutic strategy to modulate endometrial function by targeting signaling pathway(s) involving SFKs.     

Investigation on light-addressable potentiometric sensor as a possible cell-semiconductor hybrid

This article reports an investigation on light-addressable potentiometric sensor (LAPS) to be used as a possible biological cell-semiconductor hybrid that will enable us to make an interface between the physical and biological system. To increase the surface potential sensitivity, we used a LAPS structure with single insulator (SiO2) coated with poly-L-ornithine and laminin (PLOL) on Si. Efficient culturing of PC-12 and nerve cells of Lymnaea stagnalis on PLOL-coated Si3N4 and SiO2 was achieved. The thickness of the PLOL layer was found to be about 4 nm by the atomic force microscope (AFM) measurement. Using the advantage of this thin layer of PLOL, we compared the performance of a novel structure to the previously reported "PLOL-coated Si3N4/SiO2/Si" structure. Due to high insulating capacitance, the photocurrent response of the novel LAPS was found to be very steep. As a result, higher sensitivity was achieved. This steepness did not degrade during 10 days when the sensor surface was kept in contact with the cell culture medium and environment. The thickness of PLOL layer, its ability to improve the biological cell adhesion, enhanced sensitivity, and experiment with simulated neural action potential (AP) applied to the novel LAPS show a good promise for LAPS to be a biological cell-semiconductor hybrid.     

Life-cycle variation inPanonychus akitanus Ehara (Acarina:Tetranychidae)

The life history ofPanonychus akitanus Ehara was studied in two Hokkaido populations on dwarf bamboos. The Sapporo population overwintered both as egg and female adult onSasa senanensis, and the Tomakomai population overwintered as egg onSasa apoiensis. Mites of the Sapporo population produced four or five generations from late April to late November or early December. The eggs that had overwintered began to hatch in mid-May, and this hatching period overlapped with that of eggs laid in late April by females that had overwintered. Therefore, mites with different overwintering stages would interbreed. Most eggs that had overwintered in the Tomakomai population hatched in mid-May, and about four generations were produced before early December, when only eggs were found. The density of mites per leaf of the Sapporo population reached a maximum in early May on old leaves and in late June on new leaves, and thereafter gradually decreased. The Tomakomai population initially decreased in density after hatching in the spring, but rapidly dispersed to new leaves, reached a peak in early September, and then gradually decreased. The maximal density was about 10 times higher and the distribution was less clumped (lower values of the aggregation index,m/m) than that of the Sapporo population. This was probably because the Sapporo mites could utilize only the underside of sporadically distributed leaves which were curled by spiders, whereas the Tomakomai mites inhabited any part of the leaf undersurface of the hairy host plant. Predators observed were phytoseiid mites and larvae of gall midges. The predatory effect on the Sapporo population was not clear. In the Tomakomai population, the number of gall midges correlated with the number of spider mites better than that of phytoseiid mites.     

A simple quantitative chiral analysis of amino acid esters by fluorine‐19 nuclear magnetic resonance using the modified James–Bull method

A simple chiral analysis of amino acid esters by fluorine‐19 nuclear magnetic resonance (¹⁹F NMR) through the modified James–Bull method is described. Thus, amino acid ester acid salt was treated with 5‐fluoro‐2‐formylphenylboronic acid and (S)‐BINOL in the presence of triethylamine (TEA) and MS4A for 10 minutes. The reaction mixture was analysed by ¹⁹F NMR directly to afford good quantifications.     

A Combination of a Dairy Product Fermented by Lactobacilli and Galactooligosaccharides Shows Additive Effects on Mineral Balances in Growing Rats with Hypochlorhydria Induced by a Proton Pump Inhibitor

This study aimed to investigate the effects of a combination of a dairy product fermented by lactobacilli (DFL) and galactooligosaccharides (GOS) on mineral balances in growing rats with hypochlorhydria induced by a proton pump inhibitor (PPI). Three-week-old male rats were assigned to receive one of six diets: a control diet, control diets containing 1.6 or 5.0 % GOS, a DFL diet and DFL diets containing 1.6 or 5.0 % GOS for 9 days. From day 5 of the feeding period, half of the rats fed with control diets were subcutaneously administered with saline, whereas the remaining rats were administered with PPI for 5 days. Calcium (Ca), phosphorus (P), magnesium (Mg), iron (Fe) and zinc (Zn) balances were determined from days 6 to 9. PPI administration significantly decreased the apparent absorption of Ca and Fe and increased urinary P excretion, resulting in decreased Ca, Fe and P retention. GOS dose-dependently increased the apparent absorption of Ca, Mg and Fe and urinary Mg excretion and decreased urinary P excretion. DFL significantly increased the apparent absorption of Ca and Mg and urinary Mg excretion. The combination of DFL and GOS additively affected these parameters, resulting in increased Ca, P and Fe retention, and it further increased the apparent absorption and retention of Zn at 5.0 % GOS. In conclusion, the combination of DFL and GOS improves Ca, P and Fe retention in an additive manner and increases the Zn retention in growing rats with hypochlorhydria induced by PPI.     

Strength training counteracts motor performance losses during bed rest.

The purpose of the study was to determine the effect of bed rest with or without strength training on torque fluctuations and activation strategy of the muscles. Twelve young men participated in a 20-day bed rest study. Subjects were divided into a non-training group (BRCon) and a strength-training group (BRTr). The training comprised dynamic calf-raise and leg-press exercises. Before and after bed rest, subjects performed maximal contractions and steady submaximal isometric contractions of the ankle extensor muscles and of the knee extensor muscles (2.5-10% of maximal torque). Maximal torque decreased for both the ankle extensors (9%, P < 0.05) and knee extensors (16%, P < 0.05) in BRCon but not in BRTr. For the ankle extensors, the coefficient of variation (CV) for torque increased in both groups (P < 0.05), with a greater amount (P < 0.05) in BRCon (88%) compared with BRTr (41%). For the knee extensors, an increase in the CV for torque was observed only in BRCon (22%). The increase in the CV for torque in BRCon accompanied the greater changes in electromyogram amplitude of medial gastrocnemius (122%) and vastus lateralis (59%) compared with BRTr (P < 0.05). The results indicate that fluctuations in torque during submaximal contractions of the extensor muscles in the leg increase after bed rest and that strength training counteracted the decline in performance. The response varied across muscle groups. Alterations in muscle activation may lead to an increase in fluctuations in motor output after bed rest.     

Deep phylogeographical structure and parallel host range evolution in the leaf beetle Agelasa nigriceps

To understand the mechanisms behind the diversification of herbivorous insects through insect–plant interactions, it is important to know how the insects change their diet breadth in response to environmental changes. In this study, we investigated the phylogeographical pattern of the leaf beetle Agelasa nigriceps to infer the evolutionary history of its host range. While this beetle commonly uses Actinidia arguta (Actinidiaceae) as a host plant, it has been recorded recently on Pterostyrax hispidus (Styracaceae), which is now increasing in abundance at some localities in Japan due to the indirect effects of high population size of a mammalian herbivore. Considerable variation among populations in the ability of Ag. nigriceps to use P. hispidus suggests that P. hispidus is a newly acquired host plant for this beetle. Phylogenetic analyses using mitochondrial DNA sequences and amplified fragment length polymorphism (AFLP) revealed a high degree of phylogeographical structure in Ag. nigriceps throughout Japan, which is consistent with the hypothesis that several glacial refugia existed in the Japanese archipelago. In contrast, no genetic structure associated with the host plants was detected. Both the mitochondrial DNA and AFLP analyses showed that populations that can use P. hispidus are polyphyletic. These results and geographical variation in host use suggest that the host range expansion to a novel host, P. hispidus, is a very recent and possibly ongoing phenomenon and has occurred independently in several regions. Our study illustrates that the host range of herbivorous insects can evolve repeatedly in response to similar environmental changes.     

Stonogobiops yasha, a new shrimp-associated goby from Japan

A new shrimp-associated goby, Stonogobiops yasha sp. nov., is described on the basis of nine specimens collected from the Ryukyu Islands, Japan. This species is easily distinguished from other congeneric species in having reddish-orange stripes on a white body and only two median cephalic sensory pores on the head. Received: July 21, 1999 / Revised: July 10, 2001 / Accepted: July 26, 2001