Molecular characterization and expression analysis of two Na+/H+ antiporter genes in the marine red alga Porphyra yezoensis

Na⁺/H⁺ antiporters are known to play a crucial role in pH and Na⁺ homeostasis. In the present study, we characterized the molecular structures and expression patterns of two Na⁺/H⁺ antiporters from the marine red alga Porphyra yezoensis (designated PySOS1 and PyNhaD). The full-length cDNAs of PySOS1 and PyNhaD were 5122 and 1804?bp, and contained open reading frames (ORFs) of 4773 and 1275?bp, respectively. The deduced amino acid sequences showed high similarity to SOS1 and NhaD from the higher plant Arabidopsis thaliana. PySOS1 and PyNhaD contained conserved sequences found in the cation?Cproton antiporter. Quantitative real-time PCR analysis revealed that both antiporter genes were expressed in both the gametophyte and sporophyte of P. yezoensis. In addition, mRNA expression of PySOS1 and PyNhaD was simultaneously upregulated by light irradiation, suggesting that coordinated activity between the two is important in pH and Na⁺ homeostasis under light conditions. Moreover, the expression levels of both genes were partially reduced by the photosynthetic inhibitors DCMU and DBMIB, suggesting that upregulation is linked to photosynthesis-related metabolism. These findings provide an initial step towards understanding Na⁺/H⁺ antiporters in marine red algae.     

Ethanol production with <Emphasis Type="Italic">β</Emphasis>-xylosidase,xylose isomerase,and <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis> from the hydrolysate of Japanese beech after hot-compressed water treatment

Ethanol was produced from the hydrolysate collected as the water-soluble (WS) portion after hot-compressed water (HCW) treatment of Japanese beech. The process involved saccharification with β-xylosidase followed by isomerization with xylose isomerase and fermentation with Saccharomyces cerevisiae. Several process schemes were compared to investigate the effect of process integration of saccharification, isomerization, and fermentation. Higher ethanol yields were obtained for the processes that integrated isomerization and fermentation or saccharification and isomerization. Integration of isomerization and fermentation was effective in converting xylose into ethanol. Similarly, integration of saccharification and isomerization was effective in converting xylooligosaccharides into xylulose. It is presumed that the saccharification reaction toward xylose and the isomerization reaction toward xylulose were linked and therefore each reaction was enhanced.     

Demographic and height growth response of native broad-leaved deciduous tree saplings to overhead canopy release in a coastal Pinus thunbergii forest in Hokkaido, northern Japan

Prevention efforts to combat pine wilt disease must be instituted to avoid destruction of coastal forests in Hokkaido, northern Japan. We examine an alternative method involving the conversion of Pinus thunbergii forests to native broadleaf species through advancing their growth, but we have little knowledge about the response of advanced growth to overhead canopy release. To demonstrate the demographic and height growth response of advanced growth to the light change in a coastal Pinus thunbergii forest, canopy trees were removed and the sapling populations were monitored for 6?years. Inhibition in height growth caused by light increase was observed in Cerasus sargentii and Kalopanax septemlobus saplings. Especially in Kalopanax septemlobus, a remarkable decline was observed in the saplings growing in the darker positions which experienced light change. Therefore, large light changes should be carefully avoided for this species especially in the darker positions. In contrast, relative height growth rates of Quercus crispula and, especially, Quercus dentata increased with increasing light, and saplings tended to die in the darker positions. No dead trees of Sorbus commixta were observed during the study period. The saplings also showed good height growth even when under closed canopy. This species seems to be adaptive to a dark environment. Since Quercus dentata and Kalopanax septemlobus are the major components of the natural coastal forests in Hokkaido, gradual canopy release is available to foster advanced growth in coastal Pinus thunbergii forest, in accordance with the concept of density control in coastal forests.     

Reconstruction of stratum corneum in organotypically cultured canine keratinocyte-derived CPEK cells

The stratum corneum of epidermis is an essential barrier against the external environment and water loss. This study aimed to develop an organotypic culture model that targets the reconstruction of the stratum corneum using canine keratinocyte-derived CPEK cells. The CPEK cells cultured at the air-liquid interface became stratified and formed a stratum corneum-like layer on stratum spinosum- and stratum granulosum-like layers. The CPEK cells in the stratum granulosum-like layer expressed the cornified cell envelope (CCE)-related proteins loricrin and keratinocyte differentiation-associated protein. Organotypically cultured CPEK cells were considered to form a CCE at the stratum granulosum-like layer, allowing the formation of a stratum corneum-like layer. The organotypic culture of CPEK cells could be useful for studying the barrier function of canine stratum corneum.     

Involvement of neurokinin receptors in the control of pulsatile luteinizing hormone secretion in rats

It has recently been shown that neurokinin B, a tachykinin, is associated with GnRH pulse generation in sheep and goats. The aim of the present study was to clarify the role of tachykinin receptors in the control of LH secretion in rats. To this end, we evaluated the effect of CS-003, an antagonist for all three neurokinin receptors (NK1, NK2 and NK3 receptors), on pulsatile LH secretion in both sexes of rats with different routes of administration. Both oral and third ventricular administration of CS-003 suppressed LH secretion in both sexes of gonadectomized animals. Furthermore, intact male rats with oral administration of CS-003 showed decreased serum testosterone levels, which might be due to suppressed LH secretion. None of the three subtype-specific neurokinin receptor antagonists showed a significant effect on LH secretion in ovariectomized rats when each antagonist was singly administered. The present results suggest that neurokinins play a role in the control of pulsatile GnRH/LH secretion via multiple neurokinin receptors in both male and female rats.     

Isolation of a new phlorotannin, a potent inhibitor of carbohydrate-hydrolyzing enzymes, from the brown alga Sargassum patens

Ethanol extracts from 15 kinds of marine algae collected from the coast of the Noto Peninsula in Japan were examined for their inhibitory effects on human salivary α-amylase. Four extracts significantly suppressed the enzyme activity. An inhibitor was purified from the extract of Sargassum patens . The compound was a new phloroglucinol derivative, 2-(4-(3,5-dihydroxyphenoxy)-3,5-dihydroxyphenoxy) benzene-1,3,5-triol (DDBT), which strongly suppressed the hydrolysis of amylopectin by human salivary and pancreatic α-amylases. The 50% inhibitory activity (IC(50)) for α-amylase inhibition of DDBT (3.2 μg/mL) was much lower than that of commercially available α-amylase inhibitors, acarbose (26.3 μg/mL), quercetagetin (764 μg/mL), and α-amylase inhibitor from Triticum aestivum (88.3 μg/mL). A kinetic study indicated that DDBT was a competitive α-amylase inhibitor with a K(i) of 1.8 μg/mL. DDBT also inhibited rat intestinal α-glucosidase with an IC(50) value of 25.4 μg/mL for sucrase activity and 114 μg/mL for maltase activity. These results suggest that DDBT, a potent inhibitor of carbohydrate-hydrolyzing enzymes, may be useful as a natural nutraceutical to prevent diabetes.