Removal of methylene blue by invasive marine seaweed: Caulerpa racemosa var. cylindracea

Caulerpa racemosa var. cylindracea is one of the well-known invasive species in the Mediterranean Sea. In the present study, dried biomass of C. racemosa var. cylindracea was shown to have adsorption capacity for methylene blue. The adsorption reached equilibrium at 90 min for all studied concentrations (5-100mg/L). The pseudo-second-order model is well in line with our experimental results. There was a sharp increase in the adsorbed dye amount per adsorbent amount from 3.3 to 16.7 g/L, then a slight increase up to 66.7 g/L was observed. Langmuir and Freundlich's models were applied to the data related to adsorption isotherm. According to Langmuir's model data, the observed maximum adsorption capacity (qm) was 5.23 mg/g at 18 degrees C. The enthalpy of adsorption was found to be 33 kJ/mol, which indicated a chemical adsorption between dye molecules and C. racemosa var. cylindracea functional groups.     

Structure of beta-1,3-xylooligosaccharides generated from Caulerpa racemosa var. laete-virens beta-1,3-xylan by the action of beta-1,3-xylanase

Recently we reported the molecular cloning and characterization of a novel beta-1,3-xylanase from the marine bacterium Vibrio sp. AX-4 [Kiyohara et al. (2005) Biochem. J. 388, 949-957]. We report here the structural analysis of oligosaccharides generated from beta-1,3-xylan of a siphonous green alga, Caulerpa racemosa var. laete-virens, by the action of beta-1,3-xylanase. The enzyme degraded the polysaccharide producing oligosaccharides with different R(f)s on TLC (EX2-EX5). Sugar component, linkage, and MALDI-TOF-MS analyses revealed that EX2 and EX3 were Xyl-1,3-Xyl and Xyl-1,3-Xyl-1,3-Xyl, respectively. On the other hand, EX4 was a mixture of Glc-1,3-Xyl-1,3-Xyl, Xyl-1,4-Xyl-1,3-Xyl and Xyl-1,3-Xyl-1,4-Xyl, while EX5 was a mixture of tetra-saccharides containing 3-substitued Glc in addition to the same components of EX4. Branching was not likely present in EXOs prepared from the polysaccharide by the enzyme. These results strongly suggest that the C. racemosa beta-1,3-xylan is a linear heteropolysaccharide containing 1,3-Glc and 1,4-Xyl both of which are thought to be located within a beta-1,3-Xyl chain and linked via covalent bonds. This report indicates the usefulness of the enzyme for the structural analysis of beta-1,3-xylan.     

Immunomodulatory effects of Caulerpa racemosa var peltata polysaccharide and its selenizing product on T lymphocytes and NK cells in mice

A polysaccharide, CrvpPS, was isolated from Caulerpa racemosa var peltata. It was reacted with nano-selenium in distilled water containing ascorbic acid (Vit C) to form a stable CrvpPS-nano-Se complex. The immunomodulatory effects of CrvpPS and CrvpPS-nano-Se on T lymphocytes subgroups and NK cells in mice were investigated. After intragastric administration for 10 days separately, both CrvpPS and CrvpPS-nano-Se showed significant stimulatory functions to thymus gland of mice. Moreover, the CrvpPS-nano-Se induced the percentage of CD3 , CD3 CD4 , NK cells and the CD4 /CD8 value to increase significantly (P<0.05) when analyzed by flow cytometry, which is better than the CrvpPS, sucrose-nano-Se, and even the positive drug levamisole.     

Pollination Ecology and Endemic Trends in Pedicularis bracteosa var. atrosanguinea Pennell & Thompson (Scrophulariaceae) in North America

Abstract Pedicularis bracteosa var. atrosanguinea occurs locally in association with P. racemosa or P. groenlandica in the Olympic Mountains in Washington. Other plant species, e.g., Polygonum bistortoides, Lupinus argenteus var. parviflorus and Valeriana sitchensis compete for space and bumblebee pollinators within the study area. Pollinator sharing, resulting from such competition, may increase the frequency of unvisited flowers of P. bracteosa var. atrosanguinea. P. bracteosa var. atrosanguinea , with blood purple nectariferous flowers, is presumed to lack intense reflections of blue spectral components from its corollas in attracting bumblebees compared to those of P. racemosa and P. groenlandica . Six species of bumblebees ( Bombus ) and cuckoo bees ( Psithyrus ) pollinate P. bracteosa var. atrosanguinea . Of these, Bombus mixtus and B. occidentalis occur in higher frequencies and are the major pollinators of P. bracteosa var. atrosanguinea . Queen and larger worker bumblebees pollinate nototribically as they probe for nectar, while smaller worker bumblebees pollinate sternotribically while scraping pollen from anthers deeply hidden in the dome-shaped galea. Corbicular pollen loads of bumblebees collected in the study area contain Pedicularis pollen alone/in combination of Polygonum, Valeriana, Lupinus, Erigeron and Bidens , or exclusively of Polygonum or Valeriana. P. bracteosa var. atrosanguinea does not suffer seriously from deficient pollination but seedlings resulting from pollinated flowers may be subjected to natural selection pressure exerted by colonial plant species for space. P. bracteosa var. atrosanguinea does not propagate asexually but resumes aerial growth seasonally from the self-same underground root stocks. If seedlings are under continuous selection pressure for lack of space, P. bracteosa var. atrosanguinea is presumed to regenerate primarily by perennial root stocks. This behavior may favor endemism in P. bracteosa var. atrosanguinea .     

Pharmacological effects of Cimicifuga racemosa

Cimicifuga racemosa is widely employed to relieve menopause symptoms for its hormonal-like action. However, recent experimental studies have not found an estrogenic action by this plant. The purpose of this systematic review is to analyse all experimental studies (in vivo and in vitro) performed on C. racemosa to elucidate its mechanism of action. Animal and in vitro experiments on C. racemosa were identified through a computerised literature searches performed on Medline (PubMed), Embase, Amed, CISCOM and Cochrane Library databases. In addition, bibliographies of the articles thus located were scanned for further relevant publications and manufactures of commercial C. racemosa preparations were asked to contribute published and unpublished material. No language restrictions were imposed. A total of 15 animal and 15 in vitro studies on C. racemosa have been found. Their results suggest that C. racemosa possesses a central activity instead of a hormonal effect. Further biological and chemical investigations are required to define its mechanism of action and to identify the compounds responsible of its actions.     

Chemical diversity of volatiles of Teucrium orientale L. var. orientale, var. puberulens, and var. glabrescens determined by simultaneous GC-FID and GC/MS techniques

In the present work, three varieties of Teucrium orientale, var. orientale, var. puberulens, and var. glabrescens, were collected and investigated for chemical composition of the oils. Subsequent gas chromatography (GC-FID) and gas chromatography coupled to mass spectrometry (GC/MS) revealed high abundance of sesquiterpenes in the essential oils analyzed. All the oils contained β-caryophyllene (22.6, 8.5, and 6.3%, resp.) and hexadecanoic acid (7.9, 12.8, and 13.1%). Germacrene D (24.6 and 33.4%) and bicyclogermacrene (6.7 and 8.5%) were found to be the main constituents of var. orientale and var. puberulens, respectively. The high percentages of β-cubebene (26.9%), α-cubebene (9.0%), and α-copaene (7.2%) established the diversity of var. glabrescens. The qualitative difference between the essential oils allowed the differentiation between the varieties in agreement with the morphological observations described in Flora of Turkey for each variety studied. In addition, a cluster analysis of twelve Teucrium taxa based on the essential-oil composition has been carried out. Hovewer, the analysis did not clearly reflect the infrageneric classification of the genus, it largely confirmed the relationships between the infraspecific taxa of Teucrium orientale and T. chamaedrys.     

Chemical Diversity of Volatiles of Teucrium orientale L. var. orientale,var. puberulens,and var. glabrescens Determined by Simultaneous GC‐FID and GC/MS Techniques

In the present work, three varieties of Teucrium orientale, var. orientale, var. puberulens, and var. glabrescens, were collected and investigated for chemical composition of the oils. Subsequent gas chromatography (GC‐FID) and gas chromatography coupled to mass spectrometry (GC/MS) revealed high abundance of sesquiterpenes in the essential oils analyzed. All the oils contained β‐caryophyllene (22.6, 8.5, and 6.3%, resp.) and hexadecanoic acid (7.9, 12.8, and 13.1%). Germacrene D (24.6 and 33.4%) and bicyclogermacrene (6.7 and 8.5%) were found to be the main constituents of var. orientale and var. puberulens, respectively. The high percentages of β‐cubebene (26.9%), α‐cubebene (9.0%), and α‐copaene (7.2%) established the diversity of var. glabrescens. The qualitative difference between the essential oils allowed the differentiation between the varieties in agreement with the morphological observations described in Flora of Turkey for each variety studied. In addition, a cluster analysis of twelve Teucrium taxa based on the essential‐oil composition has been carried out. Hovewer, the analysis did not clearly reflect the infrageneric classification of the genus, it largely confirmed the relationships between the infraspecific taxa of Teucrium orientale and T. chamaedrys.     

Genetic maps for Pinus elliottii var. elliottii and P. caribaea var. hondurensis using AFLP and microsatellite markers

Genetic maps for individual Pinus elliottii var. elliottii and P. caribaea var. hondurensis trees were generated using a pseudo-testcross mapping strategy. A total of 329 amplified fragment length polymorphic (AFLP) and 12 microsatellite markers were found to segregate in a sample of 93 interspecfic F(1) progeny. The male P. caribaea var. hondurensis parent was more heterozygous than the female P. elliottii var. elliottii parent with 19% more markers segregating on the male side. Framework maps were constructed using a LOD 5 threshold for grouping and interval support threshold of LOD 2. The framework map length for the P. elliottii var. elliottii megagametophyte parent (1,170 cM Kosambi; 23 linkage groups) was notably smaller than the P. caribaea var. hondurensis pollen parent (1,658 cM Kosambi; 27 linkage groups). The difference in map lengths was assumed to be due to sex-related recombination variation, which has been previously reported for pines, as the difference in map lengths not be accounted for by the larger number of markers mapping to the P. caribaea var. hondurensis parent - 109 compared with 78 in P. elliottii var. elliottii parent. Based on estimated genome sizes for these species, the framework maps for P. elliottii var. elliottii and P. caribaea var. hondurensis covered 82% and 88% of their respective genomes. The pseudo-testcross strategy was extended to include AFLP and microsatellite markers in an intercross configuration. These comprehensive maps provided further genome coverage, 1,548 and 1,828 cM Kosambi for P. elliottii var. elliottii and P. caribaea var. hondurensis, respectively, and enabled homologous linkage groups to be identified in the two parental maps. Homologous linkage groups were identified for 11 out of 24 P. elliottii var. elliottii and 10 out of 25 P. caribaea var. hondurensis groups. A higher than expected level of segregation distortion was found for both AFLP and microsatellite markers. An explanation for this segregation distortion was not clear, but it may be at least in part due to genetic mechanisms for species isolation in this wide cross.     

Effect of Inula racemosa root extract on cardiac function and oxidative stress against isoproterenol-induced myocardial infarction

The cardioprotective potential of Inula racemosa root hydroalcoholic extract against isoproterenol-induced myocardial infarction was investigated in rats. The rats treated with isoproterenol (85 mg/kg, s.c.) exhibited myocardial infarction, as evidenced by significant (P < 0.05) decrease in mean arterial pressure, heart rate, contractility, relaxation along with increased left ventricular end diastolic pressure, as well as decreased endogenous myocardial enzymatic and non-enzymatic antioxidants. Isoproterenol also significantly (P < 0.05) induced lipid peroxidation and increased leakage of myocyte injury marker enzymes. Pretreatment with I. racemosa extract (50, 100 or 200 mg/kg per day, p.o.) for 21 consecutive days, followed by isoproterenol injections on days 19th and 20th significantly (P < 0.05) improved cardiac function by increasing the heart rate, mean arterial pressure, contractility and relaxation along with decreasing left ventricular end diastolic pressure. Pretreatment with I. racemosa also significantly (P < 0.05) restored the reduced form of glutathione and endogenous antioxidant enzymes superoxide dismutase, catalase, glutathione peroxidase from the heart, which were depleted after isoproterenol administration. In addition to restoration of antioxidants, I. racemosa significantly (P < 0.05) inhibited lipid peroxidation and prevented the leakage of myocytes specific marker enzymes creatine phosphokinase-MB and lactate dehydrogenase from the heart. Thus, it is concluded that I. racemosa protects heart from isoproterenol-induced myocardial injury by reducing oxidative stress and modulating hemodynamic and ventricular functions of the heart. Present study findings demonstrate the cardioprotective effect of I. racemosa and support the pharmacological relevance of its use and cardioprotection mechanism in ischemic heart disease as well as substantiate its traditional claim.     

Spatially distinct expression of two new cytochrome P450s in leaves of Nepeta racemosa/: identification of a trichome-specific isoform

Using a PCR-based approach, two novel cytochrome P450 cDNAs were isolated from a catmint (Nepeta racemosa) leaf cDNA library. The cDNAs (pBSK3C7 and pBSK4C3) were 76.9% identical in their nucleotide sequences, indicating that they are the products of two closely-related genes. A comparison of the sequence of these cDNAs with database sequences indicated that they represent new members of the CYP71 gene family of plant cytochrome P450s. Clone pBSK3C7 contains the full-length coding sequence of a cytochrome P450, whilst pBSK4C3 lacks ca. 6 codons at the 5' end. The cytochromes P450 encoded by these clones were designated CYP71A5 and CYP71A6 (pBSK3C7 and pBSK4C3, respectively). Southern blot analysis indicated that the corresponding genes were present as single copies in the genome of N. racemosa. Northern blot analysis showed that a gene homologous with CYP71A5 was expressed in the related species N. cataria, but no homologue of CYP71A6 was detected in this species. Expression of CYP71A5 in N. racemosa was maximal in flowers, tissues within the apical bud, and young expanded leaves. That of CYP71A6 was maximal in older leaves. Expression of CYP71A5 occurred exclusively in trichomes present on the leaf surfaces, in contrast to that of CYP71A6, which occurred predominantly within the leaf blade tissues.     

var1 Gene on the mitochondrial genome of Torulopsis glabrata

We have cloned and sequenced a region of the Torulopsis glabrata mitochondrial genome homologous to the Saccharomyces cerevisiae var1 gene (var1Sc). An open reading frame that could encode a protein of 339 amino acids was found with 72.7% amino acid and 85.3% nucleotide sequence homology to the S. cerevisiae var1 gene. The T. glabrata gene (var1Tg) is transcribed yielding two stable RNAs, a more abundant 13.5 S RNA and a less abundant 18 S species. We have also identified a candidate for a T. glabrata var1 protein among mitochondrial translation products labeled in isolated mitochondria. The var1Tg gene is even more A + T-rich (93%) than var1Sc (89.6%) and has conserved the strong codon bias of var1Sc. Major differences between the two sequences were found. Significant among these are that no GC clusters are found in var1Tg and the sequences surrounding each of the sites where known polymorphisms exist in var1Sc have deletions at the corresponding sites in var1Tg. These data are discussed with respect to possible origins of these var1 genes and translocation of GC clusters in S. cerevisiae mitochondrial DNA.     

Conditions and characteristics of the early stages of the sexual interaction of Pichia amethionina var. amethionina and Pichia amethionina var. pachycereana yeast cells

Pichia amethionina varieties have different sensitivity to the acidity of the medium: P. amethionina var. amethionina has the optimum pH 3 for agglutination and 5 for conjugation; P. amethionina var. pachycereana has the optimum pH 4-6 for agglutination and 6 for conjugation. The optimum temperature for the both organisms is 24 degrees C. Under the optimum conditions, P. amethionina var. amethionina has a 98% agglutination and a 28% conjugation; for P. amethionina var. pachycereana, these values are 83 and 55%, respectively. The agglutination of the two varieties does not depend on the presence of glucose. The conjugation of P. amethionina var. pachycereana does not occur in a medium without glucose while the conjugation of P. amethionina var. amethionina is reduced twofold under these conditions. Inositol, biotin and folic acid produce positive effect on the conjugation of P. amethionina var. pachycereana, but only folic acid favours the conjugation of P. amethionina var. amethionina. No differences have been found between the varieties by staining their cells and zygotes with FITC-ConA.     

低聚磷酸盐对粪产碱杆菌合成热凝胶的影响

粪产碱杆菌合成热凝胶需要消耗大量ATP用于前体物质UDPG的再生,利用3种具有不同高能磷酸键的低聚磷酸盐Na4P2O7(2P)、Na5P3O10(3P)和(NaPO3)6(6P)作为高能磷酸键供体,并取代培养基中的KH2PO4-K2HPO4(1P)而作为磷元素供体,研究其对粪产碱杆菌合成热凝胶的影响。结果表明相对于对照磷元素摩尔含量的单倍和双倍量添加3P和6P能够分别提高热凝胶产量23%和134%,达到15.1g/L和30.0g/L;同时副产物乙酸分别较对照降低了87.5%和77.7%;在以双倍量添加6P后,副产物甲酸的生成也显著降低75.7%。当培养基中不含碳酸钙进行低聚磷酸盐添加实验时,生物量显著降低,热凝胶合成几乎没有,发酵液pH最低降到2.1。以磷元素单倍量和双倍量分别添加3P和6P,并与1P混合发酵时,热凝胶产量变化不大;但是,当发酵液不存在碳酸钙而1P被作为缓冲物质时,以单倍量和双倍量添加6P使得热凝胶产量较对照分别提高60.4%和49.4%,分别达到18.4g/L和16.9g/L。     

Subtle effects of biological invasions: cellular and physiological responses of fish eating the exotic pest Caulerpa racemosa

The green alga Caulerpa racemosa var. cylindracea has invaded Mediterranean seabed including marine reserves, modifying the structure of habitats and altering the distributional patterns of associated organisms. However, the understanding of how such invasion can potentially affect functional properties of Mediterranean subtidal systems is yet to be determined. In this study, we show that C. racemosa changes foraging habit of the native white seabream, Diplodus sargus. In invaded areas, we found a high frequency of occurrence of C. racemosa in the stomach contents of this omnivorous fish (72.7 and 85.7%), while the alga was not detected in fish from a control area. We also found a significant accumulation of caulerpin, one of the main secondary metabolites of C. racemosa, in fish tissues. The level of caulerpin in fish tissues was used here as an indicator of the trophic exposure to the invasive pest and related with observed cellular and physiological alterations. Such effects included activation of some enzymatic pathways (catalase, glutathione peroxidases, glutathione S-transferases, total glutathione and the total oxyradical scavenging capacity, 7-ethoxy resorufin O-deethylase), the inhibition of others (acetylcholinesterase and acylCoA oxidase), an increase of hepatosomatic index and decrease of gonadosomatic index. The observed alterations might lead to a detrimental health status and altered behaviours, potentially preventing the reproductive success of fish populations. Results of this study revealed that the entering of alien species in subtidal systems can alter trophic webs and can represent an important, indirect mechanism which might contribute to influence fluctuations of fish stocks and, also, the effectiveness of protection regimes.     

Production of feruloyl esterases and xylanases by Talaromyces stipitatus and Humicola grisea var. thermoidea on industrial food processing by-products

Feruloyl esterase (FAE) and xylanase activities were detected in culture supernatants from Humicola grisea var. thermoidea and Talaromyces stipitatus grown on brewers' spent grain (BSG) and wheat bran (WB), two agro-industrial by-products. Maximum activities were detected from cultures of H. grisea grown at 150 rpm, with 16.9 U/ml and 9.1 U/ml of xylanase activity on BSG and WB, respectively. Maximum FAE activity was 0.47 U/ml and 0.33 U/ml on BSG and WB, respectively. Analysis of residual cell wall material after microbial growth shows the preferential solubilisation of arabinoxylan and cellulose, two main polysaccharides present in BSG and WB. The production of low-cost cell-wall-deconstructing enzymes on agro-industrial by-products could lead to the production of low-cost enzymes for use in the valorisation of food processing wastes.     

Chemical polymorphism of the essential oils from populations of Thymus caespititius grown on the island S. Jorge (Azores)

The compositions of the essential oils obtained from ten populations of Thymus caespititius grown on S. Jorge (Azores) were studied by GC and GC-MS. All the oils analysed were dominated by their monoterpene fraction (69-9%). The sesquiterpene fraction was rather small (4-17%) and consisted mainly of oxygen-containing compounds (3-13%). Enantiomeric ratio of sabinene and alpha-terpineol was investigated in the oils from some populations. (+)-Sabinene was detected with high enatiomeric purity (96 99%). Although ( +)-alpha terpineol was the predominant enantiomer, its purity varied considerably (66-97%). Cluster analysis of all identified components grouped the oils into four main clusters that corresponded with their major components: alpha-terpineol (43-68%), carvacrol (32-52%), thymol (44-58%) and sabinene (41%). The essential oil from Th. caespititius showed a clear chemical polymorphism, that was particularly evident among four populations growing along a 200 m distance at Pico Verde.     

湿加松无性系表型遗传多样性研究

以35个湿加松（Pinus elliottii×P.caribaea Morelet var.hondurensis）无性系为试验材料,对其8个表型性状进行相关性分析、聚类分析及主成分分析研究,并深入探讨了遗传多样性水平,为湿加松无性系利用及良种选育提供科学依据。结果表明：（1）生长性状变异系数介于12.31~29.28%,均值为22.90%;多样性指数介于1.61~1.80,均值为1.69。形质性状变异系数介于13.34~47.25%,均值为26.86%;多样性指数介于1.50~1.94,均值为1.77。表明湿加松无性系遗传变异较为丰富,遗传多样性水平较高。（2）8对表型性状两两间有11对呈极显著正相关,5对呈显著正相关。选定材积为该材料育种选择的主要指标,其次为树高。（3）采用系统聚类法在欧式距离为12的时候将35个无性系分为6大类群,部分来源相同或父本相同的聚为一类,说明它们在表型上的变异较小。（4）针对8个表型性状做主成分分析提取了3个主成分,累积贡献率达86.00%,主要代表了6个性状,以此来表达所有供试材料的综合变异情况。