Immunostimulatory effect by aqueous extract of <Emphasis Type="Italic">Hizikia fusiforme</Emphasis> in RAW 264.7 macrophage and whole spleen cells

Hizikia fusiforme is a commonly used food that possesses potent anti-bacterial, anti-fungal, and anti-inflammatory activities. The immunostimulatory activities of aqueous extract of Hizikia fusiforme (HFAE) in RAW 264.7 macrophages and whole spleen cells were investigated. HFAE activated RAW 264.7 macrophages to produce cytokines such as nitric oxide (NO), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6) in a dose-dependent manner. In addition, HFAE induced the mRNA expression of TNF-α, IL-1β, and IL-6 in RAW 264.7 macrophages. Moreover, HFAE stimulated proliferation of whole spleen cells and reference mitogen. Taken together, the results demonstrate that HFAE potently activates the immune function by regulating NO, TNF-α, IL-1β, and IL-6 in RAW 264.7 macrophage and promoting spleen cell proliferation.     

Triple‐Layer Structured Composite Separator Membranes with Dual Pore Structures and Improved Interfacial Contact for Sustainable Dye‐Sensitized Solar Cells

A composite separator membrane (CSM) with an A/B/A type layered structure, composed of a microporous electrolyte‐philic poly(vinylidene fluoride‐co‐hexafluoropropylene) (PVdF‐HFP) gel layer (A) and a submicrometer porous polyethylene (PE) or a macroporous poly(ethylene terephthalate) (PET) non‐woven matrix (B), is introduced in a dye‐sensitized solar cell (DSSC). Commercially available PE and PET separator membranes (SMs) act as matrices that provide mechanical stability to the DSSC and permanent pore structures for facilitated ion transport. PVdF‐HFP is used as a microporous gelator for improved interfacial contact between the solid SM and the electrodes. The PVdF‐HFP gel impedes the charge recombination process between electron and I₃ ^? at the TiO₂/electrolyte interface, resulting in improved electron lifetimes. The DSSC assembled with the CSM exhibits high initial solar energy conversion efficiency (η, 6.1%) and stable η values over 1400 h, demonstrating good long term stability. The behaviors of the DSSC are attributed to the synergistic factors of the CSM, such as improved ion conductivity, electrolyte affinity, electrolyte retention capability, effective interfacial contact, and plausible passivation of the dyes. This study demonstrates a practical combination of short‐ and long‐term DSSC performance through the introduction of the CSM.     

Nuclear magnetic resonance evidence for retention of a lamellar membrane phase with curvature in the presence of large quantities of the HIV fusion peptide

The HIV fusion peptide (HFP) is a biologically relevant model system to understand virus/host cell fusion. ²H and ³¹P NMR spectroscopies were applied to probe the structure and motion of membranes with bound HFP and with a lipid headgroup and cholesterol composition comparable to that of membranes of host cells of HIV. The lamellar phase was retained for a variety of highly fusogenic HFP constructs as well as a non-fusogenic HFP construct and for the influenza virus fusion peptide. The lamellar phase is therefore a reasonable structure for modeling the location of HFP in lipid/cholesterol dispersions. Relative to no HFP, membrane dispersions with HFP had faster ³¹P transverse relaxation and faster transverse relaxation of acyl chain ²H nuclei closest to the lipid headgroups. Relative to no HFP, mechanically aligned membrane samples with HFP had broader ³¹P signals with a larger fraction of unoriented membrane. The relaxation and aligned sample data are consistent with bilayer curvature induced by the HFP which may be related to its fusion catalytic function. In some contrast to the subtle effects of HFP on a host-cell-like membrane composition, an isotropic phase was observed in dispersions rich in phosphatidylethanolamine lipids and with bound HFP.     

Purification and Biological Activities of Enzymatically Degraded Sargassum fusiforme Polysaccharides

Enzymatic hydrolysate of the crude polysaccharide (SFP) extracted from Sargassum fusiforme was purified by column DEAE-52 and Sephadex G-100 to yield four components, namely, ESFP1, ESFP2, ESFP3 and ESFP4. These components were characterized by chemical composition assay, GC/MS, HPGPC, UV and FT-IR techniques. The in vitro antioxidant activities of the four purified fractions were investigated by measuring their radical scavenging activity and reducing power. The results suggested that all the four components possess good antioxidant activities. Among them, ESFP1 was found to possess the strongest 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and hydroxyl radical-scavenging activity, and the greatest ferric reducing power. The immunomodulatory effect of these four polysaccharides was demonstrated by their ability to promote proliferation, and to enhance both phagocytic activity and NO release in a macrophage RAW264.7 model. The results revealed that the bioactivities of the polysaccharides are related to their molecular weight, and the uronic acid and sulfate contents.     

Chemical Constituents from Sargassum fusiforme (Harv.) Setch.

Phytochemical investigation of Sargassum fusiforme (Harv.) Setch. led to the discovery of fifteen secondary metabolites, including three sterols, three monoterpenes, five nitrogenous compounds, two fatty acids, and two others. Among them, two compounds are new, while the other thirteen compounds were isolated from S. fusiforme for the first time. The structures of the two new compounds were identified by NMR and HR‐ESI‐MS data analyses, and the absolute configurations were established by comparing the calculated and experimental ECD spectroscopic data.     

Issue Information

Beautiful underwater view of an edible brown alga, Sargassum fusiforme (Fucales, Phaeophyceae) at the rocky shore in Sakurajima, Kagoshima, Japan (Photo by Ryuta Terada). Characteristic responses of the PSII photochemical efficiency on desiccation and salinity gradients in S. fusiforme are reported by Yonemori et al. in this issue.

     

Short-term variation in extracellular phosphatase activity: possible limitations for diagnosis of nutrient status in particular algal populations

Species-specific extracellular phosphatase activity (PA) was studied in natural phytoplankton with an emphasis on short-term variation in PA and its consequences for the diagnosis of inorganic phosphorus (P) deficiency. Extracellular PA was measured over 3 days at 4-h intervals to compare variations in algal PA in two distinct aquatic environments: in the Římov reservoir and acidified Plešné Lake. PA was directly detected in an epifluorescence microscope using fluorescently labelled enzyme activity; the fluorescence signal was quantified using image analysis. High short-term variation was found both in the total PA of the samples and specific PA of Ankyra ancora (G. M. Smith) Fott, Monoraphidium dybowski (Wolosz.) Hind. and Kom.-Legn., and Chlorogonium fusiforme Matwienko. The specific PA of these species varied between zero and 157 fmol cell⁻¹ h⁻¹. The PA of Chlorogonium fusiforme showed a significant relationship with the time of day. During the study period, populations of the certain species were present that differed in physiological status and activity due to diurnal or random changes caused by mixing in the reservoir or lake, or both. Therefore, species-specific extracellular PA should be interpreted carefully, as an indicator of P deficiency in freshwater phytoplankton if based on limited number of samplings.     

The effects of desiccation and salinity gradients on the PSII photochemical efficiency of an intertidal brown alga,Sargassum fusiforme from Kagoshima,Japan

The responses of photochemical efficiency to desiccation and salinity gradients in an intertidal edible brown macroalga, Sargassum fusiforme (Harvey) Setchell (Sargassaceae, Fucales), were determined using a pulse amplitude modulation (PAM)-chlorophyll fluorometer. The effective quantum yields (ΔF/F_m'; = Φ_PSII) of photosystem II (PSII) dropped to zero after 360-min aerial exposure under low irradiance (20 μmol photons m⁻² s⁻¹) and 120-min exposure under high irradiance (700 μmol photons m⁻² s⁻¹) for this species at 20°C and 50% relative humidity. Under these conditions, ΔF/F_m' failed to recover to initial levels even after 1-day rehydration in seawater. In general, ΔF/F_m' decreased as desiccation reduced the absolute water content (AWC, %). Nevertheless, when AWC was above ca. 20%, ΔF/F_m' was mostly restored to initial levels after 1-day rehydration in seawater, suggesting strong tolerance to dehydration. Furthermore, S. fusiforme appeared to tolerate a broad range of salinity (i.e. 15–50 psu) during six days of culture; however, ΔF/F_m' declined when salinity was <10 and 60 psu. Strong tolerance to dehydration and salinity stress likely provides S. fusiforme an advantage that allows it to flourish in the intertidal habitat.     

PHOTOSYNTHETIC UTILIZATION OF INORGANIC CARBON IN THE ECONOMIC BROWN ALGA,HIZIKIA FUSIFORME (SARGASSACEAE) FROM THE SOUTH CHINA SEA1

The mechanism of inorganic carbon (C_i) acquisition by the economic brown macroalga, Hizikia fusiforme (Harv.) Okamura (Sargassaceae), was investigated to characterize its photosynthetic physiology. Both intracellular and extracellular carbonic anhydrase (CA) were detected, with the external CA activity accounting for about 5% of the total. Hizikia fusiforme showed higher rates of photosynthetic oxygen evolution at alkaline pH than those theoretically derived from the rates of uncatalyzed CO₂ production from bicarbonate and exhibited a high pH compensation point (pH 9.66). The external CA inhibitor, acetazolamide, significantly depressed the photosynthetic oxygen evolution, whereas the anion‐exchanger inhibitor 4,4′‐diisothiocyano‐stilbene‐2,2′‐disulfonate had no inhibitory effect on it, implying the alga was capable of using HCO₃^? as a source of C_i for its photosynthesis via the mediation of the external CA. CO₂ concentrations in the culture media affected its photosynthetic properties. A high level of CO₂ (10,000 ppmv) resulted in a decrease in the external CA activity; however, a low CO₂ level (20 ppmv) led to no changes in the external CA activity but raised the intracellular CA activity. Parallel to the reduction in the external CA activity at the high CO₂ was a reduction in the photosynthetic CO₂ affinity. Decreased activity of the external CA in the high CO₂ grown samples led to reduced sensitiveness of photosynthesis to the addition of acetazolamide at alkaline pH. It was clearly indicated that H. fusiforme, which showed CO₂‐limited photosynthesis with the half‐saturating concentration of C_i exceeding that of seawater, did not operate active HCO₃^? uptake but used it via the extracellular CA for its photosynthetic carbon fixation.     

Photosynthetic responses of the low intertidal macroalga Sargassum fusiforme (Sargassaceae) to saline stress

Sargassum fusiforme, a species of brown seaweed with economic importance, inhabits lower intertidal zones where algae are often exposed to various stresses. In this study, changes in the photosynthetic performance of S. fusiforme under saline stress were investigated. The PSII performance in S. fusiforme significantly improved, when the thalli were exposed to 0% salinity, and remained high with prolonging treatment time. In contrast, the PSII activity declined considerably under salinities of 4.5 and 6%. The PSI activity did not change remarkably under saline stress, thus demonstrating higher tolerance to saline stress than PSII. In addition, the PSI activity could be also restored after saline treatments, when PSII was inhibited by 3-(3,4-dichlorophenyl)-1,1-dimethylurea. It might be as a result of changes in the NAD(P)H content in the thalli under saline stress. Our results suggested that PSI was much more tolerant to different saline stress than PSII in S. fusiforme. We demonstrated that S. fusiforme was much more tolerant to hyposaline than to hypersaline stress.     

MOLECULAR PHYLOGENY,ULTRASTRUCTURE, AND TAXONOMIC REVISION OF CHLOROGONIUM (CHLOROPHYTA): EMENDATION OF CHLOROGONIUM AND DESCRIPTION OF GUNGNIR GEN. NOV. AND RUSALKA GEN. NOV.1

We examined the molecular phylogeny and ultrastructure of Chlorogonium and related species to establish the natural taxonomy at the generic level. Phylogenetic analyses of 18S rRNA and RUBISCO LSU (rbcL) gene sequences revealed two separate clades of Chlorogonium from which Chlorogonium (Cg.) fusiforme Matv. was robustly separated. One clade comprised Cg. neglectum Pascher and Cg. kasakii Nozaki, whereas the other clade included the type species Cg. euchlorum (Ehrenb.) Ehrenb., Cg. elongatum (P. A. Dang.) Francé, and Cg. capillatum Nozaki, M. Watanabe et Aizawa. On the basis of unique ultrastructural characteristics, we described Gungnir Nakada gen. nov. comprising three species: G. neglectum (Pascher) Nakada comb. nov., G. mantoniae (H. Ettl) Nakada comb. nov., and G. kasakii (Nozaki) Nakada comb. nov. We also emended Chlorogonium as a monophyletic genus composed of Cg. euchlorum, Cg. elongatum, and Cg. capillatum. Because Cg. fusiforme was distinguished from the redefined Chlorogonium and Gungnir by the structure of its starch plate, which is associated with pyrenoids, we reclassified this species as Rusalka fusiformis (Matv.) Nakada gen. et comb. nov.     

Histidine Decarboxylases and Their Role in Accumulation of Histamine in Tuna and Dried Saury

Histamine-producing bacteria (HPB) such as Photobacterium phosphoreum and Raoultella planticola possess histidine decarboxylase (HDC), which converts histidine into histamine. Histamine fish poisoning (HFP) is attributable to the ingestion of fish containing high levels of histamine produced by HPB. Because freezing greatly decreases the histamine-producing ability of HPB, especially of P. phosphoreum, it has been speculated that HFP is caused by HDC itself from HPB cells autolyzing during frozen storage, even when HPB survive frozen storage. Here we constructed recombinant HDCs of P. phosphoreum, Photobacterium damselae, R. planticola, and Morganella morganii and investigated the ability of HDCs to produce sufficient histamine to cause HFP. To elucidate the character of these HDCs, we examined the specific activity of each recombinant HDC at various temperatures, pH levels, and NaCl concentrations. Further, we also investigated the stability of each HDC under different conditions (in reaction buffer, tuna, and dried saury) at various temperatures. P. damselae HDC readily produced sufficient histamine to cause HFP in fish samples. We consider that if HDC is implicated as an independent cause of HFP in frozen-thawed fish, the most likely causative agent is HDC of P. damselae.     

Fifty‐two polymorphic microsatellite loci in the rust fungus,Cronartium quercuum f.sp. fusiforme

We report on 52 microsatellite markers for use in Cronartium quercuum f.sp. fusiforme. The markers were developed from di‐, tri‐, and tetranucleotide repeat‐enriched genomic libraries. In 46 isolates collected from two natural populations in the southeastern USA, the number of alleles per locus ranged from two to 20 (mean 6.94) with gene diversity values ranging from 0.043 to 0.933 (mean 0.537). The markers should prove highly useful for genetic ‘fingerprinting’ of single‐spore isolates commonly used in host–pathogen gene interaction studies, as marker loci for linkage mapping studies, and for examining fine‐scale population genetic structure in natural populations of the fungus.     

Immunostimulatory Effect of Enzyme-Modified <Emphasis Type="Italic">Hizikia fusiforme</Emphasis>in a Mouse Model In Vitro and Ex Vivo

Hizikia fusiforme, a brown seaweed, has been utilized as a health food and in traditional medicine. In this study, we investigated whether enzyme-modified H. fusiforme extracts (EH) have immunological effects compared with normal H. fusiforme extracts (NH). The effects of NH and EH on immune responses were investigated by assessing nitric oxide (NO) production, phagocytosis, and cytokine secretion in RAW 264.7 murine macrophages and mice. Also, fucosterol was evaluated to find the active component of NH and EH by addressing cytotoxicity test and NO production. Both of NH and EH significantly increased cell viability and NO synthesis. Tumor necrosis factor-α (TNF-α) expression was more induced by EH with LPS treatment. Phagocytic activity, as the primary function of macrophages, was markedly induced by EH treatment. Additionally, EH encouraged splenocyte proliferation and recovered the levels of cytokines IL-1β, IL-6, and TNF-α in mice. Finally, fucosterol increased NO production with no cytotoxicity, which means that fucosterol is an active component of EH. In conclusion, EH has the potential to modulate immune function and could offer positive therapeutic effect for immune system diseases.     

Fusobacterium nucleatum subsp. fusiforme Gharbia and Shah 1992 is a Later Synonym of Fusobacterium nucleatum subsp. vincentii Dzink et al. 1990

On the basis of the DNA–DNA hybridization patterns and phenotypic characteristics, Fusobacterium nucleatum was classified into five subspecies. Previous studies have suggested that F. nucleatum subsp. vincentii is genetically similar to F. nucleatum subsp. fusiforme. The aim of this study was to investigate the possibility of classifying these two subspecies into a single subspecies by phylogenetic analysis using a single sequence (24,715 bp) concatenated 22 housekeeping genes of eight F. nucleatum strains including type strains of five F. nucleatum subspecies. The phylogenetic analysis indicated that F. nucleatum subsp. vincentii and F. nucleatum subsp. fusiforme were clustered in the same group and each strain of other F. nucleatum subspecies were also separated into the same cluster. These results suggested that F. nucleatum subsp. fusiforme and F. nucleatum subsp. vincentii can be classified into a single subspecies. F. nucleatum subsp. vincentii was early published name; therefore, F. nucleatum subsp. fusiforme Gharbia and Shah 1992 can be regarded as a later synonym of F. nucleatum subsp. vincentii Dzink et al. 1990.     

Surface ultrastructure and molecular phylogenetics of four unarmored heterotrophic dinoflagellates, including the type species of the genus Gyrodinium (Dinophyceae)

Small subunit rRNA gene sequences were determined for four unarmored heterotrophic dinoflagellates (Gyrodinium spirals, the type species of the genus Gyrodinium, as well as G. fusiforme, Gymnodinium rubrum and the freshwater species G. helveticum) using a single‐cell polymerase chain reaction (PCR) technique. For identification and record keeping, each cell was carefully observed and photographed using a light microscope under high magnification, prior to single‐cell PCR. G. rubrum and G. helveticum possess an elliptical apical groove and longitudinal striations similar to those of G. spirale and G. fusiforme, and molecular phylogenetic studies reveal that the four species form a single clade. We therefore propose the following new combinations: Gyrodinium rubrum (Kofoid et Swezy) Takano et Horiguchi comb. nov. and Gyrodinium helveticum (Penard) Takano et Horiguchi comb. nov.     

Effects of hypo- and hypersalinity on photosynthetic performance of Sargassum fusiforme (Fucales,Heterokontophyta)

Photoprotection mechanisms protect photosynthetic organisms, especially under stress conditions, against photodamage that may inhibit photosynthesis. We investigated the effects of short-term immersion in hypo- and hypersalinity sea water on the photosynthesis and xanthophyll cycle in Sargassum fusiforme (Harvey) Setchell. The results indicated that under moderate light [110 μmol(photon) m^?2 s^?1], the effective quantum yield of PSII was not reduced in S. fusiforme fronds after 1 h in hyposalinity conditions, even in fresh water, but it was significantly affected by extreme hypersalinity treatment (90‰ sea water). Under high light [HL, 800 μmol(photon) m^?2 s^?1], photoprotective mechanisms operated efficiently in fronds immersed in fresh water as indicated by high reversible nonphotochemical quenching of chlorophyll fluorescence (NPQ) and de-epoxidation state; the quantum yield of PSII recovered during the subsequent relaxation period. In contrast, fronds immersed in 90‰ sea water did not withstand HL, barely developed reversible NPQ, and accumulated little antheraxanthin and zeaxanthin during HL, while recovery of the quantum yield of PSII was severely inhibited during the subsequent relaxation period. The data provided concrete evidence supporting the short-term tolerance of S. fusiforme to immersion in fresh water compared to hypersalinity conditions. The potential practical implications of these results were also discussed.     

Equilibrium and Kinetic Studies of Cd2+ Biosorption by the Brown Algae Sargassum fusiforme

A fundamental investigation of the biosorption of Cd²⁺ from aqueous solution by the edible seaweed Sargassum fusiforme was performed under batch conditions. The influences of experimental parameters, such as the initial pH, sorption time, temperature, and initial Cd²⁺ concentration, on Cd²⁺ uptake by S. fusiforme were evaluated. The results indicated that the biosorption of Cd²⁺ depended on the initial Cd²⁺ concentration, as well as the pH. The uptake of Cd²⁺ could be described by the Langmuir isotherm model, and both the Langmuir biosorption equilibrium constant and the maximum biosorption capacity of the monolayer decreased with increasing temperature, thereby confirming the exothermic character of the sorption process. The biosorption kinetics follows the pseudo-second-order kinetic model, and intraparticle diffusion is the sole rate-limiting step for the entire biosorption period. These fundamental equilibrium and kinetic results can support further studies to the removal of cadmium from S. fusiforme harvested from cadmium-polluted waters.