The immunomodulating effect of phlorotannin was investigated in mice stimulated by ovalbumin. When analyzing the main components of phlorotannin concentrate (PTC) from Eisenia nipponica, seven phlorotannins [eckol, 6,6′-bieckol, 6,8′-bieckol, 8,8′-bieckol, dieckol, phlorofucofuroeckol (PFF)-A, and PFF-B] were detected. These phlorotannins accounted for approximately 80% of PTC. Oral administration of PTC to mice daily for 21 days reduced serum immunoglobulin E (IgE) and total IgG1 levels attributable to Th2 cells. The production of splenic cytokines [interleukin (IL)-10 and transforming growth factor-β1] and Treg cell-mediated expression of forkhead box protein P3 mRNA were significantly increased whereas the production of inflammatory cytokines (interferon-γ, IL-4, IL-5, and IL-17) by Th1, Th2, and Th17 cells was markedly suppressed. IL-21 production and basic leucine zipper ATF-like transcription factor mRNA expression attributable to follicular helper T (Tfh) cells were also suppressed. Flow cytometric analyses demonstrated increased number of Treg cells despite a decrease in the total T cell population. An increase in total B cells was also observed by the flow cytometric analyses in addition to increases in IL-10 production, which activates B cells. In contrast, the significantly suppressed production of inflammatory cytokines and moderate increase in Treg cell subpopulation indicated a direct impact of PTC on inflammatory lymphocytes (Th1, Th2, Th17, and Tfh). Thus, PTC may exert antiallergic effects by immunomodulation of T cells and inactivation of inflammatory lymphocyte.

Because the number of people suffering from allergies has significantly increased, improved ways of treating these conditions by medical, pharmaceutical, and dietary means are required. Large numbers of studies on allergy have been conducted, and many anti-allergic compounds have been found. Phenolic compounds from terrestrial plants, including catechins and flavonoids, possess anti-allergic properties. Although polyphenols are present in some brown algae, their anti-allergic activities were not studied in detail before the 1990s. The focus was on the algal polyphenols, collectively called phlorotannins (eg., eckol, 6,6′-bieckol, 8,8′-bieckol, dieckol, and phlorofucofuroeckol-A), and research was conducted to clarify their anti-allergic activities. This review summarizes the anti-allergic effects of phlorotannins isolated from the brown alga, Eisenia nipponica, and related reports by other research groups.

Phlorotannin is the collective term for polyphenols derived from brown algae belonging to the genera Ascopyllum, Ecklonia, Eisenia, Fucus and Sargassum etc. Since the incidence of allergies is currently increasing in the world, there is a focus on phlorotannins having anti-allergic and anti-inflammatory effects. In this study, six purified phlorotannins (eckol; 6,6′-bieckol; 6,8′-bieckol; 8,8′-bieckol; phlorofucofuroeckol (PFF)-A and PFF-B) from Eisenia arborea, orally administered to mice, were examined for their suppression effects on ear swelling. In considering the suppression, we also examined whether the phlorotannins suppressed release of chemical mediators (histamine, leukotriene B4 and prostaglandin E2), and mRNA expression and/or the activity of cyclooxygenase-2 (COX-2), using RBL-2H3 cells, a cultured mast cell model. Results showed that the phlorotnannins exhibited suppression effects in all experiments, with 6,8′-bieckol, 8,8′-bieckol and PFF-A showing the strongest of these effects. In conclusion, orally administered phlorotannins suppress mouse ear swelling, and this mechanism apparently involves suppression of chemical mediator release and COX-2 mRNA expression or activity. This is the first report of the anti-allergic effects of the orally administered purified phlorotannins in vivo. Phlorotannins show potential for use in functional foods or drugs.

Ammonium and nitrate are the primary nitrogen sources in natural environments, and are essential for growth and development in photosynthetic eukaryotes. In this study, we report on the isolation and characterization of an ammonium transporter gene (PyAMT1) which performs a key function in nitrogen (N) metabolism of Pyropia yezoensis thalli. The predicted length of PyAMT1 was 483 amino acids (AAs). The AA sequence included 11 putative transmembrane domains and showed approximately 33-44% identity to algal and plant AMT1 AA sequences. Functional complementation in an AMT-defective yeast mutant indicated that PyAMT1 mediated ammonium transport across the plasma membrane. Expression analysis showed that the PyAMT1 mRNA level was strongly induced by N-deficiency, and was more highly suppressed by resupply of inorganic-N than organic-N. These results suggest that PyAMT1 plays important roles in the ammonium transport system, and is highly regulated in response to external/internal N-status.

In flowering plants, floral homeotic MADS-box genes, which constitute a large multigene family, play important roles in the specification of floral organs as defined by the ABCDE model. In this study, a MADS-box gene, ZjMADS1, was isolated and characterized from the marine angiosperm Zostera japonica. The predicted length of the ZjMADS1 protein was 246 amino acids (AA), and the AA sequence was most similar to those of the SEPALLATA (SEP) subfamily, corresponding to E-function genes. Southern blot analysis suggested the presence of two SEP3-like genes in the Z. japonica genome. ZjMADS1 mRNA levels were extremely high in the spadices, regardless of the developmental stage, compared to other organs from the reproductive and vegetative shoots. These results suggest that the ZjMADS1 gene may be involved in spadix development in Z. japonica and act as an E-function gene in floral organ development in marine angiosperms.

Molecular chaperones are synthesized and accumulated under a variety of unfavorable conditions in all organisms. Heat shock protein 90 (Hsp90) and Hsp60, which are classified into the major classes of molecular chaperones, play important roles in cellular stress responses. In this study, we characterized sterile Ulva pertusa Hsp90 (UpHsp90) and UpHsp60 genes which may be involved in tolerance to thermal and heavy metal stresses in this alga. The UpHsp90 cDNA consisting of 2,118 nucleotides encoded a polypeptide of 705 amino acids (AA). On the other hand, the UpHsp60 cDNA consisting of 1,722 nucleotides encoded a protein whose predicted length was 573 AA. The AA sequence alignment and phylogenetic analyses showed that the UpHsp90 and UpHsp60 proteins were more similar to cytoplasmic Hsp90s and mitochondrial Hsp60s, respectively, than to other types of the respective Hsps. Southern blot analysis indicated that the sterile U. pertusa genome had at least two cytoplasmic Hsp90-encoding genes and two mitochondrial Hsp60-encoding genes. The UpHsp90 and UpHsp60 mRNA levels were significantly affected by diurnal and temperature changes, and slightly affected by exposure to heavy metals. These results suggest that UpHsp90 and UpHsp60 genes play particularly important roles in adaptation to diurnal and temperature changes.

Synthesis and accumulation of molecular chaperones are universal responses found in all cellular organisms when exposed to a variety of unfavorable conditions. Heat shock protein 70 (Hsp70), which is one of the major classes of molecular chaperones, plays a particularly important role in cellular stress responses, and the Hsp70 system is the most intensely studied in higher plants and algae. Therefore, we isolated and characterized a cDNA clone encoding Hsp70 from a sterile strain of Ulva pertusa (Ulvales, Chlorophyta). The sterile U. pertusa Hsp70 (UpHsp70) cDNA consisted of 2,272 nucleotides and had an open reading frame encoding a polypeptide of 663 amino acid (AA) residues with a molecular mass of 71.7 kDa. Amino acid alignment and phylogenetic analysis of Hsp70s from other organisms showed that UpHsp70 was more similar to cytoplasmic Hsp70s from green algae and higher plants (≥75%) than to those from other algae and microorganisms. Southern blot analysis indicated that the sterile U. pertusa genome had at least four cytoplasmic Hsp70-encoding genes. UpHsp70 mRNA levels were significantly affected by diurnal changes, rapidly increased by high-temperature stress, and gradually increased by exposure to copper, cadmium, and lead. These results suggest that UpHsp70 plays particularly important roles in adaptation to high-temperature conditions and diurnal changes, and is potentially involved in tolerance to heavy metal toxicity.

We isolated two different genomic DNAs (UprbcS1 and UprbcS2) encoding the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase and portions of the 5′- and 3′-flanking regions from sterile Ulva pertusa Kjellman. The UprbcS1 and UprbcS2 genes had three introns in the coding region. Each predicted UprbcS polypeptide was a 180-amino-acid (AA) residue including a 38-AA transit peptide, although the 104th AA residue was replaced. The nucleotide sequences of UprbcS cDNAs isolated from a cDNA library corresponded to that of the UprbcS1 gene, suggesting that the UprbcS1 gene was predominantly expressed in sterile U. pertusa compared to UprbcS2. Southern blot analysis showed that each UprbcS gene was a single-copy gene in the sterile U. pertusa genome. Northern hybridization indicated that the expression of UprbcS was induced and repressed by dark and light treatments, respectively. When sterile U. pertusa cells were transformed with an expression vector containing the UprbcS1 promoter and terminator sequences fused with the green fluorescent protein (GFP) gene, GFP fluorescence was observed in the cells transformed. These results suggest that the UprbcS1 gene promoter is light regulated and highly active in the sterile U. pertusa cells and is available for genetic transformation system in the alga.

The anti-allergic effect of an edible brown alga, Eisenia arborea, was investigated by determining the anti-degranulation activity on RBL cells or KU812 cells and the inhibitory effects on enzymatic activities involved in eicosanoid (leukotoriene and prostaglandin) synthesis. When the stimulated cells were treated with a M/C (methanol:chloroform=1:2, v/v) extract of the alga, the histamine release of the cells was suppressed in a dose-dependent manner, and eicosanoid release from the cells was also strongly inhibited. Furthermore, the M/C extract and six phlorotannins (eckol, 6,6′-bieckol, 6,8′-bieckol, 8,8′-bieckol, phlorofucofuroeckol-A, phlorofucofuroeckol-B) derived from the extract inhibited enzymatic activities responsible for eicosanoid synthesis. This is the first report of the enzymatic activity inhibition by the phlorotannins 6,6′-bieckol, 6,8′-bieckol and PFF-B. Since histamine and eicosanoids are known to mediate the allergic reaction, inhibition of these mediators and enzymatic activities could be responsible for the anti-allergic effect of the alga.

The nutritive value of Capsosiphon fulvescens (C. Agardh) Setchell et Gardner, a new developing species for cultivation of marine macroalgae in Korea, was assessed by comparison with common edible green seaweed Ulva prolifera (Oeder) J. Agardh collected from Korea and Japan, based on analysis of its chemical composition. The contents of protein and of total, essential, and free amino acids of C. fulvescens were significantly higher than those of U. prolifera, whereas those of lipid, carbohydrate, and total dietary fibres were significantly lower. There were no significant differences in the moisture content between Capsosiphon and Ulva species. The main minerals of Capsosiphon and Ulva are Na, Mg, K, Ca, and Al, making up approximately 97–99 % of the total minerals. The contents of Na and V of Capsosiphon were significantly greater than those of Ulva, while those of Mn and Sr were significantly lower in Capsosiphon. The contents of retinal (27.8 μg g−1 dry wt) and ascorbic acid (0.28 mg g−1 dry wt) of Capsosiphon were significantly higher than those of Ulva, but the contents of cobalamin were lower. These results suggest that C. fulvescens has greater potential to be used as human food and as an ingredient in formulated food.

The rhodophyte seaweed Porphyra yezoensis, known more commonly world-wide as "nori", is an important commercial crop in Japan. Cultivation of nori in Japan is often affected by outbreaks of "iroochi", a discoloration of the thalli due to a decrease in inorganic nutrients in the culture area that in turn decreases the amount of photosynthetic pigments in the thalli. Treating thalli with inorganic nitrogen can reverse iroochi. In this paper, we report on the characterization of three P. yezoensis genes, a nitrate transporter (PyNRT2) and two urea transporters (PyUT1 and PyUT2), which may be involved in reversing iroochi. The predicted length of the PyNRT2 protein was 479 amino acids (AA), while PyUT1 and PyUT2 were 740 and 680 AA, respectively. PyNRT2 was more similar to NRT2 from a chromophyte than to NRTs from Chlamydomonas and higher plants. The two P. yezoensis UTs had 56% AA identity to each other, and showed the closest relationship to higher plant and yeast DUR3 proteins which formed a subfamily of the sodium-solute symporter protein family. Hydrophobicity plots of the AA sequences showed that the PyNRT2, PyUT1, and PyUT2 included 12, 15, and 16 transmembrane domains, respectively. Southern blot analysis indicated that the P. yezoensis genome has a single NRT2-encoding gene and at least four UT-encoding genes. Expression analysis of PyNRT2 and PyUT genes showed that the messenger RNA level of the PyNRT2 gene reached a maximum after 48 h in the nitrate starvation condition and was then restored to the constitutive level, while expression of the PyUT genes was induced in proportion to treatment times in the nitrate starvation condition. These results suggest that the PyNRT2 and PyUT are responsible for the high-affinity nitrate/urea transport systems that operate under low external nitrate concentrations.

To investigate the anti-allergic effects of the brown alga Eisenia arborea. A strain of Brown Norway rats know to strongly respond to immunoglobulin E (IgE) were used as an allergy model animal. The rats were immunized with ovalbumin by oral administration. The levels of serum IgE and histamine were suppressed in the rats fed a diet supplemented with dried E. arborea powder. As for the cytokine pattern, the interferon-γ production in the spleens and mesenteric lymph nodes (MLN) was enhanced, and the interleukin-4 (IL-4) production in the spleens and/or IL-10 production in the spleens and MLN were suppressed. These results, together with the change in the Th1/Th2 balance, indicate that the rats fed with E. arborea became more anti-allergic, suggesting that E. arborea might possess anti-allergic effects.

Six anti-allergic phlorotannins from the brown alga Eisenia arborea were examined for their inhibitory effects on oxidation and hyaluronidase activities. To investigate the radical scavenging activities of the phlorotannins, the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity was evaluated. Although the radical scavenging activity of a typical scavenger, epigallocatechin gallate (EGCg), was the strongest, the six tested phlorotannins exhibited activities of various degrees. When the hyaluronidase inhibitory activity was examined, eckol inhibited the hyaluronidase activity similar to the typical inhibitors, EGCg and disodium cromoglycate. The other five phlorotannins inhibited the enzymatic activity stronger than the above inhibitors. Since the six phlorotannins from E. arborea expressed radical scavenging and hyaluronidase inhibitory activities, E. arborea could be used as an anti-inflammatory and anti-allergic food ingredient.

Eight commercial samples of 'nori', or the dried laver Porphyra spp. were extracted with 75% ethanol and the extracts were analyzed for free sugars and organic acids by gas chromatography and for minerals by atomic absorption spectrophotometry. Isofloridoside and floridoside were found to be the predominant free sugars, followed by glucose and galactose. A small amount of mannose, inositol, ribose, arabinose and xylose were also detected. The contents of these sugars were generally lower in nori products of good quality than in those of poor quality. Volatile and nonvolatile organic acids found in the extracts were formic, acetic, propionic, butyric, lactic, oxalic, malonic, fumaric, succinic, malic and citric acids. Among them, formic acid was found to vary greatly from sample to sample. The contents of these organic acids were generally lower in tasty nori samples. Major cations found were Na+ and K+, followed by Mg2+ and Cat+, whereas major anions were Cland PO43-. The nori produced from the indoor cultured laver showed a general pattern similar to nori samples of poor quality, as far as the above components are concerned.

Gametogenesis of Porphyra yezoensis thalli is induced by ageing as well as by changing water temperature and photoperiod. Under laboratory conditions, thalli cultivated at 10 ∘C with a 10:14 h (light: dark) cycle develop vegetatively to adult form without gametogenesis. On the other hand, sexual reproduction, which involves differentiation of vegetative cells and subsequent gametogenesis, is induced by culturing at 15 ∘C with a 16: 8 h (light: dark) cycle. We have constructed subtracted cDNA libraries enriched for differentially expressed transcripts in vegetative and reproductive thalli, and randomly selected 1,152 cDNAs from each subtracted library. Results of the dot blot analyses used for identification of differentially expressed cDNAs indicated that mRNA levels of 176 and 138 cDNAs tended to increase in the vegetative and reproductive thalli, respectively. BLAST analysis of nucleotide and deduced amino acid sequences showed that the cDNAs represented 63 and 59 unique clones for the vegetative and reproductive cDNA enriched subtracted libraries, respectively. Interestingly, some of the cDNAs isolated from the reproductive subtracted library were homologous to genes encoding protein kinases, GTP-binding protein, and heat shock proteins involved in signal transduction and the molecular chaperon system.

Pythium porphyrae is a fungal pathogen responsible for red rot disease of the seaweed Porphyra (Rhodophyta). Infection forecasts of Porphyra by P. porphyrae were estimated from the epidemiological observations of Porphyra thalli and numbers of zoospore of P. porphyrae in laboratory and cultivation areas. Four features of forecasting infections were determined by relating zoospore concentrations to the incidence of thallus infection; infection (in more than 1000 zoospores L −1 ), microscopic infection [less than 2 mm in diameter of lesion (in from 2000 to 3000 zoospores L −1 )], macroscopic infection [more than 2 mm in diameter of lesion (in from 3000 to 4000 zoospores L −1 ), and thallus disintegration (in more than 4000 zoospores L −1 ). High zoospore concentrations led to more infection. The tendency that zoospore concentration of P. porphyrae increased with the rate of infection of Porphyra thalli was generally observed in forecasting infections in both the laboratory and in cultivation areas. Based on the Porphyra cultivation areas, the accuracy and consistency of forecasting infections suggest that this method could be employed to manage and control red rot disease.

The effects of 80% methanol extracts from frozen samples of 41 macroalgae and one sea grass collected in the Ise-Shima region of Japan were investigated on histamine release from rat basophile leukemia cells (RBL-2H3) sensitized with antidinitrophenyl (DNP) IgE and stimulated with DNP-BSA. Of the 21 brown algae, five green and 15 red algae, and one sea grass tested, only extracts from seven brown algae suppressed histamine release from RBL cells, as determined by highperformance liquid chromatography (HPLC) analysis. When the cytotoxic effects of the seven brown algal extracts were investigated by Trypan blue staining, only Eisenia arborea and Sargassum thunbergil did not show cytotoxic effects. Therefore, we conclude that E. arborea and S. thunbergii may contain compounds that have antiallergic effects without inducing cell death.