RESUMEN
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-ß1] 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.
Asunto(s)
Phaeophyceae , Linfocitos T , Animales , Citocinas , Ratones , Ovalbúmina , ARN MensajeroRESUMEN
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.
Asunto(s)
Ciclooxigenasa 2/metabolismo , Inflamación/tratamiento farmacológico , Phaeophyceae/química , Taninos/farmacología , Administración Oral , Animales , Línea Celular , Relación Dosis-Respuesta a Droga , Oído , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Inflamación/inducido químicamente , Ratones , Ratones Endogámicos ICR , Estructura Molecular , ARN Mensajero/genética , ARN Mensajero/metabolismo , Transducción de Señal , Taninos/administración & dosificación , Taninos/químicaRESUMEN
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.
Asunto(s)
Compuestos de Amonio/metabolismo , Nitrógeno/metabolismo , Rhodophyta/fisiología , Rhodophyta/genéticaAsunto(s)
Virus de la Parainfluenza 1 Humana/aislamiento & purificación , Virus de la Parainfluenza 2 Humana/aislamiento & purificación , Virus de la Parainfluenza 3 Humana/aislamiento & purificación , Virus de la Parainfluenza 4 Humana/aislamiento & purificación , Infecciones por Respirovirus/epidemiología , Infecciones por Rubulavirus/epidemiología , Adolescente , Adulto , Anticuerpos Antivirales/sangre , Niño , Preescolar , Femenino , Pruebas de Inhibición de Hemaglutinación , Humanos , Lactante , Japón/epidemiología , Masculino , Persona de Mediana Edad , ARN Viral/genética , ARN Viral/aislamiento & purificación , Infecciones por Respirovirus/virología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Infecciones por Rubulavirus/virología , Estudios Seroepidemiológicos , Adulto JovenRESUMEN
Reverse transcription polymerase chain reaction (RT-PCR) and real-time RT-PCR were used to detect 14 (6.6%) influenza C virus (InfC) among 213 clinical samples collected from children with respiratory symptoms in Mie Prefecture, Japan, between January 2012 and December 2012. Virus isolation using Madin-Darby canine kidney cells and/or embryonated chicken eggs was also successful for 3 of the 14 PCR-positive samples. Eleven patients (78.6%) were aged <3 years. Phylogenetic analysis of the hemagglutinin-esterase gene showed that the InfC detected in Mie Prefecture belonged to the C/Sao Paulo/82-related lineage. To determine the seroprevalence of InfC, a total of 575 serum samples from patients aged 1 month to 69 years in Mie Prefecture were screened by hemagglutination inhibition test using the C/Mie/199/2012 (C/Sao Paulo/82-related lineage) strain as the antigen. The samples with an antibody titer of ≥1:16 were designated as antibody-positive. The results showed that 53.7% of the 296 serum samples collected in 2011 and 85.3% of the 279 samples collected in 2012 were positive for antibodies against InfC, suggesting that an outbreak of InfC infection occurred in Mie Prefecture in 2012. Therefore, continuous and proactive monitoring is important to determine the number of InfC-infections and to better understand the epidemiology.
Asunto(s)
Gammainfluenzavirus/clasificación , Gammainfluenzavirus/genética , Gripe Humana/epidemiología , Gripe Humana/virología , Adolescente , Adulto , Anciano , Animales , Anticuerpos Antivirales/sangre , Niño , Preescolar , Análisis por Conglomerados , Femenino , Glicoproteínas Hemaglutininas del Virus de la Influenza/genética , Humanos , Lactante , Gammainfluenzavirus/aislamiento & purificación , Japón/epidemiología , Masculino , Persona de Mediana Edad , Datos de Secuencia Molecular , Filogenia , ARN Viral/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Análisis de Secuencia de ADN , Homología de Secuencia de Aminoácido , Estudios Seroepidemiológicos , Adulto JovenRESUMEN
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.
Asunto(s)
Regulación de la Expresión Génica de las Plantas , Proteínas de Dominio MADS/genética , Proteínas de Dominio MADS/metabolismo , Zosteraceae/genética , Zosteraceae/metabolismo , Secuencia de Aminoácidos , Flores/crecimiento & desarrollo , Dosificación de Gen , Orden Génico , Proteínas de Dominio MADS/química , Datos de Secuencia Molecular , Filogenia , Proteínas de Plantas/química , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Alineación de Secuencia , Homología de Secuencia de Aminoácido , Zosteraceae/clasificaciónRESUMEN
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 (> or =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.
Asunto(s)
ADN Complementario/química , Proteínas HSP70 de Choque Térmico/genética , Ulva/genética , Secuencia de Aminoácidos , ADN Complementario/aislamiento & purificación , Fertilidad , Expresión Génica/efectos de los fármacos , Proteínas HSP70 de Choque Térmico/metabolismo , Metales Pesados/toxicidad , Datos de Secuencia Molecular , ARN Mensajero/metabolismo , Estrés Fisiológico , Transcripción Genética/efectos de los fármacos , Ulva/metabolismoRESUMEN
Eisenia arborea is an edible brown alga occasionally used as a folk medicine in gynecopathy in Japan. A new phlorotannin was isolated from the alga during our search for naturally occurring anti-allergic compounds from edible algae guided by the inhibitory effect on histamine release from rat basophile leukemia (RBL)-2H3 cells. The phlorotannin was called "phlorofucofuroeckol-B." Its structure was determined by spectral analysis and chemical conversion. This paper describes the isolation, structure elucidation, and inhibitory effect of phlorofucofuroeckol-B on histamine release.
Asunto(s)
Antialérgicos/química , Antialérgicos/farmacología , Benzofuranos/química , Benzofuranos/farmacología , Dioxinas/química , Dioxinas/farmacología , Laminaria/química , Animales , Antialérgicos/aislamiento & purificación , Benzofuranos/aislamiento & purificación , Línea Celular Tumoral , Dioxinas/aislamiento & purificación , Alimentos , Histamina/metabolismo , Espectroscopía de Resonancia Magnética , Estructura Molecular , RatasRESUMEN
We constructed a cDNA library from sterile Ulva pertusa (Ulvales, Chlorophyta), and isolated and characterized a full-length cDNA clone encoding actin. The actin (ACT) cDNA consisted of 1487 nucleotides (nt) and had an open reading frame (ORF) encoding a polypeptide of 377 amino acid (AA) residues. The ACT gene had one intron in the 5'-untranslated region and three introns in the coding region. Transcription started 26 nt downstream of the putative TATA box. A potential polyadenylation signal, TGTAG, was located 100 nt downstream of the terminator codon, TAG. Amino acid alignment with actins from various algae and land plants showed that sterile U. pertusa actin was more similar to actins from Chlorophyta, Phaeophyta, Euglenophyta, and higher plants (over 76.9%) than to actins from Rhodophyta. Southern blot analysis indicated that the sterile U. pertusa genome has only a single actin-encoding gene. Thalli grown on a 12D/12L photoperiod increased in surface area some two-fold over 24 h regardless of the nutritional conditions. The growth rate of thalli during the light period was significantly higher than that during the dark period. Northern hybridization indicated that the expression of actin mRNA was induced and repressed by the light and dark treatments, respectively. These results suggest that the U. pertusa cell division cycle has a periodicity of 24 h and that the ACT gene is highly transcribed during cell growth and development in the light period.
Asunto(s)
Actinas/genética , Ulva/genética , Secuencia de Aminoácidos , Secuencia de Bases , Northern Blotting , Medios de Cultivo/farmacología , ADN/química , ADN/genética , ADN/aislamiento & purificación , Dosificación de Gen , Expresión Génica/efectos de los fármacos , Genes/genética , Datos de Secuencia Molecular , Mutación , Fotoperiodo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Alineación de Secuencia , Análisis de Secuencia de ADN , Homología de Secuencia de Aminoácido , Ulva/crecimiento & desarrollo , Ulva/efectos de la radiaciónRESUMEN
Sterile Ulva, which is a macroalga, has the potential to grow stably; therefore, this seaweed is expected to be an efficient resource of functional food containing various nutrients such as sulfur amino acids, proteins, carbohydrates, and minerals. Ulva latuca was selected from the "Marine Park" in Tokyo Bay, and its growth rate (g-dry/[m2.d]) was measured using model reactors located on the land or on the surface of the sea at Yokohama. The growth rate of U. lactuca was recorded to be approx 20 g-dry/(m2.d), which is estimated to be 10 times greater than that in a natural field in the Marine Park. In addition, this growth rate was higher than that of conventional crops such as corn and rice on a farm or paddy. These data led us to newly design and propose a floating type of labor-efficient U. lactuca production system. d-Cysteinolic acid, which is included in U. lactuca as a major sulfur amino acid, inhibited the Fenton reaction, resulting in suppression of hydroxyl radical production and singlet oxygen. Addition of the sulfur amino acid (1 microM) to HepG2 cells markedly decreased the intracellular triglyceride level. Hence, this proposed facility also has the potential for industrial production of a valuable resource for the primary prevention of lifestyle-related diseases using enriched or eutrophied seawater.