Roosting ecology of endangered plant‐roosting bats on Okinawa Island: Implications for bat‐friendly forestry practices

Roosting information is crucial to guiding bat conservation and bat‐friendly forestry practices. The Ryukyu tube‐nosed bat Murina ryukyuana (Endangered) and Yanbaru whiskered bat Myotis yanbarensis (Critically Endangered) are forest‐dwelling bats endemic to the central Ryukyu Archipelago, Japan. Despite their threatened status, little is known about the roosting ecology of these species and the characteristics of natural maternity roosts are unknown. To inform sustainable forestry practices and conservation management, we radio‐tracked day roosts of both species in the subtropical forests of Okinawa''s Kunigami Village District. We compared roost and roost site characteristics statistically between M. ryukyuana nonmaternity roosts (males or nonreproductive females), maternity roosts, and all M. yanbarensis roosts. Generalized linear models were used to investigate roost site selection by M. ryukyuana irrespective of sex and age class. Lastly, we compiled data on phenology from this and prior studies. Nonreproductive M. ryukyuana roosted alone and primarily in understory foliage. Murina ryukyuana maternity roosts were limited to stands >50 years old, and ~60% were in foliage. Myotis yanbarensis roosted almost entirely in cavities along gulch bottoms and only in stands >70 years old (~1/3 of Kunigami''s total forest area). Murina ryukyuana maternity roosts were higher (4.3 ± 0.6 m) than conspecific nonmaternity roosts (2.3 ± 0.5 m; p < .001) and M. yanbarensis roosts (2.7 ± 0.5 m; not significant). Model results were inconclusive. Both species appear to be obligate plant roosters throughout their life cycle, but the less flexible roosting preferences of M. yanbarensis may explain its striking rarity. To conserve these threatened bats, we recommend the following forestry practices: (a) reduce clearing of understory vegetation, (b) refrain from removing trees along streams, (c) promote greater tree cavity densities by protecting old‐growth forests and retaining snags, and (d) refrain from removing trees or understory between April and July, while bats are pupping.     

Involvement of the cytoplasmic juxtamembrane region of matriptase in its exclusive localization to the basolateral membrane domain of Madin–Darby canine kidney epithelial cells

Matriptase is a type II transmembrane serine protease. This protease is strongly expressed in simple epithelial cells such as enterocytes and kidney tubular cells in which the plasma membranes are separated into apical and basolateral domains. Although matriptase was found previously to occur exclusively on the basolateral membrane of enterocytes, the underlying mechanism of localization is unclear. In the present study, a full-length rat matriptase and a chimera consisting of the cytoplasmic and transmembrane regions of the protease and green fluorescent protein (designated as 1–86GFP) were found to localize exclusively to the basolateral membrane domain when expressed in Madin–Darby canine kidney epithelial cells. Mutagenesis analysis of 1–86GFP revealed that the matriptase cytoplasmic juxtamembrane amino acid residues (Lys45, Val47, and Arg50) play a role in mediating the localization in the cells. This study provides the first evidence that matriptase carries information for its localization in simple epithelia.     

Isolation and characterization of polymorphic microsatellite DNA markers in the Japanese eight‐barbel loach,Lefua echigonia

We isolated and characterized 17 polymorphic microsatellite loci for the Japanese eight‐barbel loach, Lefua echigonia, an endangered freshwater species in streams including agricultural canals in Japan. The number of observed alleles for each locus ranged from two to nine, and the values of observed and expected heterozygosities ranged from 0.125 to 0.844 and from 0.148 to 0.876, respectively. All loci conformed to Hardy–Weinberg equilibrium. Genetic linkage disequilibrium analysis between all pairs of the loci showed no significant departure from the null hypothesis between any of the loci.     

ENZYME ACTIVITIES ASSOCIATED WITH RIBOSOMES FROM SOYBEAN SEEDLINGS

Ribosomes from cotyledons of soybean seeds soaked overnightin water (resting-cotyledon ribosomes) and respective ribosomesfrom cotyledons (working-cotyledon ribosomes) and hypocotyls(hypocotyl ribosomes) of 5 day-old seedlings were prepared.These ribosomes mainly consisted of 80 S particles. However,hypocotyls contained 115 S particles in a small amount and working-cotyledonscontained 115 and 150 S particles which may correspond to thepolymers of ribosomes or polysomes. The abundant 80 S ribosomeswere fractionated by the sucrose density gradient centrifugation,and enzyme activities in the fractionated ribosomes were estimated.RNase, PDase, acid phosphatase, 5'-nucleotidase, XTPase, peroxidaseand ß-glucosidase were found in the ribosomes. Theenzyme activities were lower in the resting-cotyledon ribosomesand higher in the hypocotyl ribosomes. Working-cotyledon ribosomesshowed the middle of them. All these enzymes were also foundin the cytoplasmic solution (supernatant) of cotyledon and hypocotylcells. However, RNase, PDase, 5'-nucleotidase and peroxidasewere concentrated in ribosomes, and the specific activitiesof 5'-nucleotidase and ß-glucosidase were increasedby washing the ribosomes. The status of the enzymes found inthe ribosomes was discussed. (Received May 12, 1966; )     

Identification of potent lysophosphatidic acid receptor 5 (LPA5) antagonists as potential analgesic agents

Lysophosphatidic acid (LPA) plays an important role in a variety of cellular functions. In particular, LPA5 receptor is highly expressed in spinal cord and dorsal root ganglion, which are associated with pain. This fact prompted us to hypothesize that LPA5 antagonists show analgesic effects. To search for potent LPA5 antagonists with blood brain barrier (BBB) permeability, we conducted high throughput screening (HTS). In HTS campaign, we found a 2H-isoquinoline-1-one scaffold showing antagonistic activity against LPA5 and synthesized a series of 2H-isoquinoline-1-one derivatives and evaluated their LPA5 activities. Among these compounds, compound 7e showed potent LPA5 activity with an IC₅₀ value of 0.12?μM, and acceptable BBB permeability. Furthermore, it showed effective analgesic effect in a chronic constriction injury rat model. Therefore, 7e may have a potential as novel pain therapeutic approach.     

Resolution ofFusarium sporotrichioides proteins by two-dimensional polyacrylamide gel electrophoresis and identification by sequence homology comparison in protein data base

Proteins fromFusarium sporotrichioides M-1-1, a T2-toxin-producing strain, were separated by two-dimensional polyacrylamide gel electrophoresis. One thousand two hundred and forty-four protein spots were resolved and 103 protein spots were subjected to N-terminal sequencing. Fifty-eight protein spots were sequenced and 48 proteins were observed to have blocked N termini. Forty out of 58 sequenced proteins were identified by homology search against the PIR protein sequence data base and protein superfamily data base, while the residual 18 sequences were not identified. Twenty-seven of the N-terminal-blocked proteins were subjected to mild anhydrous hydrazine vapor deblocking. Twenty-four spots were not deblocked indicating the presence of acyl groups at the N termini, while 3 proteins were deblocked showing the blocked group to be pyrroglutamyl carboxylic acid residues. The results can provide a more global view of cellular genetic expression than any other technique. The created data may offer a unique opportunity to link information with DNA sequence data.