Design and synthesis of opioidmimetics containing 2',6'-dimethyl-L-tyrosine and a pyrazinone-ring platform

Twelve 2',6'-dimethyl-L-tyrosine (Dmt) analogues linked to a pyrazinone platform were synthesized as 3- or 6-[H-Dmt-NH(CH(2))(n)],3- or 6-R-2(1H)-pyrazinone (n=1-4). 3-[H-Dmt-NH-(CH(2))(4)]-6-beta-phenethyl-5-methyl-2(1H)-pyrazinone 11 bound to mu-opioid receptors with high affinity (K(i)mu=0.13 nM; K(i)delta/K(i)mu=447) with mu-agonism (GPI IC(50)=15.9 nM) and weak delta-antagonism (MVD pA(2)=6.35). Key factors affecting opioid affinity and functional bioactivity are the length of the aminoalkyl chain linked to Dmt and the nature of the R residue. These data present a simplified method for the formation of pyrazinone opioidmimetics and new lead compounds.     

Identification of 6-substituted 4-arylsulfonyl-1,4-diazepane-2,5-diones as a novel scaffold for human chymase inhibitors

A novel series of 6-substituted 4-sulfonyl-1,4-diazepane-2,5-diones were designed, synthesized and evaluated as human chymase inhibitors. Structure-activity relationship studies led to the identification of a potent inhibitor, (6S)-6-(5-chloro-2-methoxybenzyl)-4-[(4-chlorophenyl)sulfonyl]-1,4-diazepane-2,5-dione, with an IC(50) of 0.027 microM.     

Synthesis and SAR studies of a novel class of S1P1 receptor antagonists

A series of Sodium 4-[(4-butoxyphenyl)thio]-2'-substituted-1,1'-biphenyl-3- sulfonates were identified as functional sphingosine-1-phosphate (S1P) antagonists with selectivity for the S1P(1) receptor subtype starting from chemical lead 2, which was found while screening our in-house compound library. We performed chemical modifications on each regional structure of compound 2, for example, on the three ring compartments, the benzyl substituents, and the long alkyl chain part. The introduction of a biphenyl skeletal structure and the installation of a hydroxyl group onto the terminal carbon in the side-chain region resulted in the potent derivative 35c, which showed >500-fold more potent S1P(1) inhibitory activity than lead compound 2. We report herein the synthesis and structure-activity relationships of structurally novel S1P(1) receptor antagonists.     

Tissue specific expression of the splice variants of the mouse vacuolar proton-translocating ATPase a4 subunit

We have identified splicing variants of the mouse a4 subunit which have the same open reading frame but have a different 5′-noncoding sequence. Further determination of the 5′-upstream region of the a4 gene in mouse indicated the presence of two first exons (exon 1a and exon 1b) which include the 5′-noncoding sequence of each variant. The mRNAs of both splicing variants (a4-I and a4-II) show a similar expression pattern in mouse kidney by in situ hybridization. However, tissue and developmental expression patterns of the variants are different. In addition to strong expression in kidney, a4-I expression was detected in heart, lung, skeletal muscle, and testis, whereas a4-II is expressed in lung, liver, and testis. During development, a4-I was expressed beginning with the early embryonic stage, but a4-II mRNA was detected from day17. These results suggest that each a4 variant has both a tissue and developmental stage specific function.     

Right-elevated expression of charon is regulated by fluid flow in medaka Kupffer's vesicle

Recent studies have revealed that a cilium-generated liquid flow in the node has a crucial role in the establishment of the left-right (LR) axis in the mouse. In fish, Kupffer's vesicle (KV), a teleost-specific spherical organ attached to the tail region, is known to have an equivalent role to the mouse node during LR axis formation. However, at present, there has been no report of an asymmetric gene expressed in KV under the control of fluid flow. Here we report the earliest asymmetric gene in teleost KV, medaka charon, and its regulation. Charon is a member of the Cerberus/DAN family of proteins, first identified in zebrafish. Although zebrafish charon was reported to be symmetrically expressed in KV, medaka charon displays asymmetric expression with more intense expression on the right side. This asymmetric expression was found to be regulated by KV flow because symmetric and up-regulated charon expression was observed in flow-defective embryos with immotile cilia or disrupted KV. Taken together, medaka charon is a reliable gene marker for LR asymmetry in KV and thus, will be useful for the analysis of the early steps downstream of the fluid flow.     

Feeding rates and absorption efficiencies of four species of sea urchins (genus Echinometra) fed a prepared diet

Four closely related species of sea urchins belonging to the genus Echinometra, Echinometra sp. A (Ea), E. mathaei (Em), E. sp. C (Ec), and E. oblonga (Eo), occur sympatrically but in different microhabitats on Okinawan coral reefs. Feeding rates and absorption efficiencies of the four species were investigated in the laboratory by feeding sea urchins ad libitum a diet prepared from turf algae and agar over a 7-day period. Feeding rates differed significantly among the four species of Echinometra (Ea > Em approximately = Ec > Eo). Absorption efficiencies of protein and lipid did not differ significantly among the four species. Carbohydrate, a major nutrient component in the diet, was absorbed by the four species at significantly different efficiencies (Eo > Ec approximately = Em > Ea), which resulted in similar interspecific differences in absorption efficiencies of dry matter, total organic matter, and energy. The amount of nutrients absorbed from the diet was directly related to the feeding rate, indicating that the increase in absorption efficiency was not sufficient to completely compensate for low feeding rate. The interspecific difference in physiological performance in relation to feeding and absorption is consistent with taxonomic differentiation among the four species.     

Selection of rub trees by brown bears (Ursus arctos) in Hokkaido, Japan

Tree rubbing by brown bears (Ursus arctos) is a well-known behavior throughout the animal’s distribution. There is still insufficient information on the characteristics and function of the behavior. We investigated seasonal frequency of tree rubbing by brown bears, characteristics and reasons for selection of rub trees, and characteristics of bear signs on and around rub trees in a mixed coniferous–broad-leaved deciduous forest in Hokkaido, Japan. Between 1998 and 2009, we found 172 rub trees and confirmed 995 tree rubbings. We found that the rub trees were used repeatedly by bears within a year and for multiple years (more than 10 years). Tree rubbing by brown bears was observed from April to November, with a peak between May and June that corresponds to the mating season of brown bears. Abies sachalinensis was selected and broad-leaved trees were avoided for tree rubbing based on estimated availability in natural forest. The preference for Abies sachalinensis might be because the strong odor of resin attracts bears for rubbing their head and neck in resinous substances and for increasing the detectability of their markings by receptor bears. Selected trees for rubbing were located right beside the trail on relatively level ground among trees along roads or trails. Trees had a relatively large diameter at breast height. These characteristics would also serve to increase the access and detectability of their markings. Series of pad-shaped depressions was the most frequently observed (70 %) indication of bear rubbing, followed by debarking (51 %) and clawing (30 %). In terms of visual and olfactory signal amplification, physical damage by bears to the trees might have a function to enhance the smell as a result of increase in outflow of the resin. We conclude that tree rubbing behavior is associated with the mating season of brown bears and that the main purpose of this behavior is scent marking to communicate intraspecifically during the mating season.