Organotin Compounds (OTCs) in Saccharina latissima (Phaeophyceae) from the Barents Sea

Doklady Biological Sciences - The degree of contamination of Saccharina latissima, the dominant species among macrophyte algae in the sublittoral zones of the Kola Bay and Eastern Murman of the...     

The complete amino acid sequence of trypsin inhibitor DE-3 from Erythrina latissima seeds

Trypsin inhibitor DE-3 from Erythrina latissima seeds contains 172 amino acids, including 4 half-cystine residues, and resembles the Kunitz-type inhibitors. Limited hydrolysis of DE-3 with trypsin at pH 3 produced two fragments, F1 and F2, containing 63 and 109 amino acids, respectively. Amino-terminal sequence studies revealed that F1 was the N-terminal and that F2 was the C-terminal fragment. The complete amino acid sequence of fragments F1 and F2 was then determined on peptides produced by enzymatic digestion with trypsin and Staphylococcus aureus V8 protease. The sequence of trypsin inhibitor DE-3 from E. latissima seeds shows a high degree of homology to those of Kunitz-type trypsin inhibitors from soybeans and winged bean seeds.     

中国台湾布丹蛛属1新种（蜘蛛目：巨蟹蛛科）

本文报道了蜘蛛目巨蟹蛛科布丹蛛属1新种：宽大布丹蛛Bhutaniella latissima sp. nov.该标本采自台湾岛屏东县大汉山自然保护区,标本保存于湖北大学生命科学学院动物行为与进化中心。     

Purification and some properties of two proteinase inhibitors (DE-1 and DE-3) from Erythrina latissima (broad-leaved erythrina) seed

Two proteinase inhibitors, DE-1 and DE-3, were purified from Erythrina latissima seeds. Whereas DE-1 inhibits bovine chymotrypsin and not bovine trypsin, DE-3 inhibits trypsin but not chymotrypsin. The molecular weights and the amino acid compositions of the two inhibitors resemble the corresponding properties of the Kunitz-type proteinase inhibitors. The N-terminal primary structure of DE-3 showed homology with soybean trypsin inhibitor (Kunitz) and also with the proteinase inhibitors (A-II and B-II) from Albizzia julibrissin seed.     

Antimicrobial and radical scavenging flavonoids from the stem wood of Erythrina latissima

From the stem wood of Erythrina latissima, two isoflavones and a flavanone were isolated and characterized as 7,3'-dihydroxy-4'-methoxy-5'-(gamma,gamma-dimethylallyl)isoflavone (erylatissin A), 7,3'-dihydroxy-6',6'-dimethyl-4',5'-dehydropyrano [2',3': 4',5']isoflavone (erylatissin B), (-)-7,3'-dihydroxy-4'-methoxy-5'-(gamma,gamma-dimethylallyl)flavanone (erylatissin C), respectively, in addition to 10 known flavonoids. Structures of these compounds were determined on the basis of their spectroscopic data. These compounds showed antimicrobial activity against Escherichia coli, Staphylococcus aureus, Bacillus subtilis and Candida mycoderma. The isolated compounds also exhibited weak radical scavenging properties towards DPPH radical.     

CHEMICAL CHANGES IN LESPEDEZA ASSOCIATED WITH INDUCED POLYPLOIDY

Hanson , C. H. (U.S.D.A., Beltsville, Md.), W. A. Cope and R. M. Brinkley . Chemical changes in Lespedeza associated with induced polyploidy. Amer. Jour. Bot. 46(1): 36-39. 1959.— Changes in the quantities of 5 chemical constituents which were associated with induced polyploidy were studied using leaves of the following polyploids: Autotetraploids—Lespedeza cuneata, L. latissima, L. stipulacea, and L. striata; amphidiploids—L. hedysaroides × L. cuneata and L. latissima × L. cuneata. L. stipulacea and L. striata were annuals with low concentrations of tannin; the other species were perennials and high in tannin content. The polyploids were generally lower in crude protein, P₂O₅, and CaO, and higher in K₂O and total tannin. The greatest and most significant shift was in total tannin. Induced polyploids of Lespedeza did not appear promising as forage plants.     

Amino acid sequence of winged bean (Psophocarpus tetragonolobus (L.) DC.) chymotrypsin inhibitor, WCI-3

The complete amino acid sequence of winged bean chymotrypsin inhibitor 3 (WCI-3) was determined by the conventional methods. WCI-3 consisted of 183 amino acid residues, but was heterogeneous in the carboxyl terminal region owing to the loss of one to four carboxyl terminal amino acid residues. The sequence of WCI-3 was highly homologous with those of soybean trypsin inhibitor Tia, winged bean trypsin inhibitor WTI-1, and Erythrina latissima trypsin inhibitor DE-3. One of the reactive site peptide bonds of WCI-3 was identified as Leu(65)-Ser(66), which was located at the same position as those of the other Kunitz-family leguminous proteinase inhibitors.     

Purification of human tissue plasminogen activator with Erythrina trypsin inhibitor

Seeds of the legume Erythrina latissima contain a 20,000-dalton, single-chain protein that has been shown to inhibit the amidolytic activity of trypsin and tissue plasminogen activator. It had no comparable effect on urokinase. IC50 values of 1.1 X 10(-7) M for tissue plasminogen activator and 6.9 X 10(-10) M for trypsin were determined by titration. When coupled to agarose, the Erythrina inhibitor provided an effective reagent for affinity purification of tissue plasminogen activator from melanoma cell-conditioned tissue culture medium. Using this as a single-step procedure, 270-fold purified enzyme was reproducibly obtained with yields of 90% or greater. Both one- and two-chain forms of tissue plasminogen activator were purified. The enzyme migrated, in sodium dodecyl sulfate-polyacrylamide gel electrophoresis, as a predominant 72,000-dalton doublet with lesser amounts of immunochemically similar, 115,000- and 68,000-dalton components.     

The reactive sites of proteinase inhibitors from Erythrina seeds

Although the Kunitz-type proteinase inhibitors from the seeds of various Erythrina species have similar molecular weights (approximately 20,000), and share many other chemical characteristics, they could nevertheless be divided into three groups on the basis of their relative abilities to inhibit chymotrypsin, trypsin and tissue plasminogen activator. Group a inhibitors were relatively specific for chymotrypsin; they were poor inhibitors of trypsin and had no apparent effect upon tissue plasminogen activator. Group b proteins inhibited trypsin strongly and chymotrypsin slightly less effectively. They had no effect upon t-PA. Group c inhibitors inhibited trypsin, chymotrypsin and t-PA. Analysis of the amino acid composition of the three groups of inhibitors revealed major differences in alanine content. Minor differences in the content of most other amino acids were also noticed. Group b and group c inhibitors had, in most cases, the same reactive sites (Arg-Ser). The sequences neighbouring the reactive sites showed a significant degree of homology. Chemical modification of arginine in proteinase inhibitors from the seeds of E. latissima and soybeans using 1-2-cyclohexanedione confirmed the presence or absence of arginine in the reactive sites.     

Altered kelp (Laminariales) phlorotannins and growth under elevated carbon dioxide and ultraviolet-B treatments can influence associated intertidal food webs

Due to the importance of brown algae, such as kelp (Laminariales, Phaeophyta), within most cool nearshore environments, any direct responses of kelp to multiple global changes could alter the integrity of future coastal marine systems. Fifty-five-day manipulation of carbon dioxide (CO₂) and ultraviolet light (UVB) within outdoor sea-tanks, approximating past, present and two predicted future levels, examined the direct influences on Saccharina latissima (= Laminaria saccharina ) and Nereocystis luetkeana development and biochemistry, as well as the indirect influences on a marine herbivore ( Tegula funebralis ; Gastropoda, Mollusca) and on naturally occurring intertidal detritivores. Kelp species displayed variable directional (negative and positive growth) and scale responses to CO₂ and UVB manipulations, which was influenced by interactions. Kelp phlorotannin (phenolic) production in blade tissues was induced by elevated UVB levels, and especially enhanced (additively) by elevated CO₂, further suggesting that some actively growing kelp species are carbon limited in typical nearshore environments. Negative indirect effects upon detritivore consumers fed CO₂-manipulated kelp blade tissues were detected, however, no statistical relationships existed among UVB-treated tissues, and test herbivores did not distinguish between phlorotannin-altered CO₂: UVB-treated kelp blade tissues. Results suggest that past and future conditions differentially benefit these kelp species, which implies a potential for shifts in species abundance and community composition. Higher CO₂ conditions can indirectly impede marine decay processes delaying access to recycled trace nutrients, which may be disruptive to the seasonal regrowth of algae and/or higher trophic levels of nearshore ecosystems.     

Seasonal sea ice cover as principal driver of spatial and temporal variation in depth extension and annual production of kelp in Greenland

We studied the depth distribution and production of kelp along the Greenland coast spanning Arctic to sub‐Arctic conditions from 78 ºN to 64 ºN. This covers a wide range of sea ice conditions and water temperatures, with those presently realized in the south likely to move northwards in a warmer future. Kelp forests occurred along the entire latitudinal range, and their depth extension and production increased southwards presumably in response to longer annual ice‐free periods and higher water temperature. The depth limit of 10% kelp cover was 9–14 m at the northernmost sites (77–78 ºN) with only 94–133 ice‐free days per year, but extended to depths of 21–33 m further south (73 ºN–64 ºN) where >160 days per year were ice‐free, and annual production of Saccharina longicruris and S. latissima, measured as the size of the annual blade, ranged up to sevenfold among sites. The duration of the open‐water period, which integrates light and temperature conditions on an annual basis, was the best predictor (relative to summer water temperature) of kelp production along the latitude gradient, explaining up to 92% of the variation in depth extension and 80% of the variation in kelp production. In a decadal time series from a high Arctic site (74 ºN), inter‐annual variation in sea ice cover also explained a major part (up to 47%) of the variation in kelp production. Both spatial and temporal data sets thereby support the prediction that northern kelps will play a larger role in the coastal marine ecosystem in a warmer future as the length of the open‐water period increases. As kelps increase carbon‐flow and habitat diversity, an expansion of kelp forests may exert cascading effects on the coastal Arctic ecosystem.     

The structure of biogenic habitat and epibiotic assemblages associated with the global invasive kelp <Emphasis Type="Italic">Undaria pinnatifida</Emphasis> in comparison to native macroalgae

Kelp forests dominate temperate and polar rocky coastlines and represent critical marine habitats because they support elevated rates of primary and secondary production and high biodiversity. A major threat to the stability of these ecosystems is the proliferation of non-native species, such as the Japanese kelp Undaria pinnatifida (‘Wakame’), which has recently colonised natural habitats in the UK. We quantified the abundance and biomass of U. pinnatifida on a natural rocky reef habitat over 10 months to make comparisons with three native canopy-forming brown algae (Laminaria ochroleuca, Saccharina latissima, and Saccorhiza polyschides). We also examined the biogenic habitat structure provided by, and epibiotic assemblages associated with, U. pinnatifida in comparison to native macroalgae. Surveys conducted within the Plymouth Sound Special Area of Conservation indicated that U. pinnatifida is now a dominant and conspicuous member of kelp-dominated communities on natural substrata. Crucially, U. pinnatifida supported a structurally dissimilar and less diverse epibiotic assemblage than the native perennial kelp species. However, U. pinnatifida-associated assemblages were similar to those associated with Saccorhiza polyschides, which has a similar life history and growth strategy. Our results suggest that a shift towards U. pinnatifida dominated reefs could result in impoverished epibiotic assemblages and lower local biodiversity, although this could be offset, to some extent, by the climate-driven proliferation of L. ochroleuca at the poleward range edge, which provides complex biogenic habitat and harbours relatively high biodiversity. Clearly, greater understanding of the long-term dynamics and competitive interactions between these habitat-forming species is needed to accurately predict future biodiversity patterns.     

Assessing species richness of macrofauna associated with macroalgae in Arctic kelp forests (Hornsund,Svalbard)

Kelp beds support diverse and productive benthic coastal ecosystems and are often perceived as cold water analogs of tropical coral reefs, yet the levels of species richness in polar regions have remained largely unexplored. The present study aims to assess the magnitude of macrozoobenthic species richness associated with the macroalgae in an Arctic kelp forest. The study was conducted in Hornsund, a high latitude Arctic fjord off west Spitsbergen (76–77°N). A total of 403 samples (i.e., individual algae) were collected by scuba divers at three sites located along the fjord axis at varying depths (5–10 m). The most common invertebrate species and the sample species richness (mean 11.5 species per sample) were consistent among the most common algal species (Laminaria digitata, Saccharina latissima, Alaria esculenta, Desmarestia aculeta, Odonthalia dentata, Phycodrys rubens). Fauna associated with overstory kelps was concentrated on the holdfasts. A total of 208 species were identified; Bryozoa, Polychaeta and Hydrozoa yielded the highest numbers of species (70, 52 and 37, respectively). The non-parametric Chao2 estimator of true species richness gave an estimate of 259 species (with 95% confidence intervals from 234 to 308). Despite the high sampling effort, the total species richness was not captured. This may stem from the high level of rarity within the fauna associated with kelps; 38% of the species occurred only in one or two samples. The studied fauna was much less diverse than similar assemblages surveyed at lower latitudes. The high numbers of species and the common occurrence of colonial forms (Bryozoa, Hydrozoa) in the current study show that these taxa cannot be omitted in macrobenthic diversity surveys performed within kelp habitats.     

Fucans, but not fucomannoglucuronans, determine the biological activities of sulfated polysaccharides from Laminaria saccharina brown seaweed

Sulfated polysaccharides from Laminaria saccharina (new name: Saccharina latissima) brown seaweed show promising activity for the treatment of inflammation, thrombosis, and cancer; yet the molecular mechanisms underlying these properties remain poorly understood. The aim of this work was to characterize, using in vitro and in vivo strategies, the anti-inflammatory, anti-coagulant, anti-angiogenic, and anti-tumor activities of two main sulfated polysaccharide fractions obtained from L. saccharina: a) L.s.-1.0 fraction mainly consisting of O-sulfated mannoglucuronofucans and b) L.s.-1.25 fraction mainly composed of sulfated fucans. Both fractions inhibited leukocyte recruitment in a model of inflammation in rats, although L.s.-1.25 appeared to be more active than L.s.-1.0. Also, these fractions inhibited neutrophil adhesion to platelets under flow. Only fraction L.s.-1.25, but not L.s.-1.0, displayed anticoagulant activity as measured by the activated partial thromboplastin time. Investigation of these fractions in angiogenesis settings revealed that only L.s.-1.25 strongly inhibited fetal bovine serum (FBS) induced in vitro tubulogenesis. This effect correlated with a reduction in plasminogen activator inhibitor-1 (PAI-1) levels in L.s.-1.25-treated endothelial cells. Furthermore, only parent sulfated polysaccharides from L. saccharina (L.s.-P) and its fraction L.s.-1.25 were powerful inhibitors of basic fibroblast growth factor (bFGF) induced pathways. Consistently, the L.s.-1.25 fraction as well as L.s.-P successfully interfered with fibroblast binding to human bFGF. The incorporation of L.s.-P or L.s.-1.25, but not L.s.-1.0 into Matrigel plugs containing melanoma cells induced a significant reduction in hemoglobin content as well in the frequency of tumor-associated blood vessels. Moreover, i.p. administrations of L.s.-1.25, as well as L.s.-P, but not L.s.-1.0, resulted in a significant reduction of tumor growth when inoculated into syngeneic mice. Finally, L.s.-1.25 markedly inhibited breast cancer cell adhesion to human platelet-coated surfaces. Thus, sulfated fucans are mainly responsible for the anti-inflammatory, anticoagulant, antiangiogenic, and antitumor activities of sulfated polysaccharides from L. saccharina brown seaweed.     

Effect of supplementing sheep diets with macroalgae species on in vivo nutrient digestibility,rumen fermentation and blood amino acid profile

In this study, a brown macroalgae species, Saccharina latissima, processed to increase its protein concentration, and a red macroalgae species, Porphyra spp., were used to evaluate their in vivo digestibility, rumen fermentation and blood amino acid concentrations. Four castrated rams were used, whose diets were supplemented with a protein-rich fraction of S. latissima, a commercial Porphyra spp. and soybean meal (SBM). Our results show that the protein digestibility of a diet with S. latissima extract was lower (0.55) than those with Porphyra spp. (0.64) and SBM (0.66). In spite of the higher nitrogen (N) intake of diets containing Porphyra spp. and SBM (20.9 and 19.8 g N/day, respectively) than that with S. latissima (18.6 g N/day), the ratio of N excreted in faeces to total N intake was significantly higher in the diet with S. latissima than those with Porphyra spp. and SBM. This reflects that the utilization of protein in S. latissima was impaired, possibly due to reduced microbial activity. The latter statement is corroborated by lower volatile fatty acid composition (25.6, 54.8 and 100 mmol/l for S. latissima, Porphyra spp. and SBM, respectively) and a non-significant tendency for lower ammonia concentration observed in diets with S. latissima and Porphyra spp. compared to SBM. It is important to note that the S. latissima used in this trial was rinsed during processing to remove salt. This process potentially also removes other water-soluble compounds, such as free amino acids, and may have increased the relative fraction of protein resistant to rumen degradation and intestinal absorption. Furthermore, the phlorotannins present in macroalgae may have formed complexes with protein and fibre, further limiting their degradability in rumen and absorption in small intestines. We recommend that further studies explore the extent to which processing of macroalgae affects its nutritive properties and rumen degradability, in addition to studies to measure the intestinal absorption of these macroalgae species.     

Rapid phosphorylation of MAP kinase-like proteins in two species of Arctic kelps in response to temperature and UV radiation stress

Mitogen-activated protein kinases (MAPKs) are a group of cytoplasmic phosphoproteins that constitute the central core of the signalling network to respond to stress in most organisms. Their role in stress responses has been extensively studied in organisms from yeast to humans, and recently, their presence has also been described in higher plants as well as in micro- and macroalgae. In this study, we demonstrate via short experiments (1 h in duration), the rapid activation of two MAPKs similar to p38 and JNK of mammalian cells, in the Arctic kelps Laminaria solidungula and Saccharina latissima exposed to temperature and UV stress. The molecular mass of p38 is 40 kDa in L. solidungula and 42 kDa in S. latissima, while two JNKs were detected in both species, of 36 and 42 kDa in L. solidungula, and 36 and 40 kDa in S. latissima. These MAPKs are highly phosphorylated in response to temperature and UV light. In S. latissima, both p38 and the JNK showed higher phosphorylation at 2 °C than at 7 °C, while the reverse response was shown for L. solidungula. In addition, a significant increase in phosphorylation of both kinases was found following exposure to UV radiation (UVR). Exposure to PAR + UVA + UVB induced higher phosphorylation than PAR + UVA in L. solidungula, especially at 7 °C. In S. latissima, this response occurred only with JNK, and no differences in p38 phosphorylation between PAR + UVA and PAR + UVA + UVB at any temperature were observed. These results indicate the possible participation of MAPK-like proteins in response to stress in Arctic kelps, and that their activation is species-specific.     

Anthropology and genetics of the caucasus peoples and the problem of the origin of the caucasoids

The electrophoretical polymorphisms of some blood proteins were studied in the Talysh population of Pirasora situated in South-East Azerbaidjan. We calculated the gene frequencies of these polymorphisms and determined the genetic distances between the Talyshes and some Iranian populations of North, Central and South Iran, Afghans, and three populations of Azerbaijan. The Talyshes are very close to Iranians of Shiraz, whereas they are distant from the Azerbaijanians. Anthropological investigations showed that the Caucasoids and Mongoloids lived in the Aragvi Basin since the Eneolithic period. This was stated by Alexeev (1974), who emphasized the mixture of the Caucasus populations from ancient times on. We calculated the genetic distances between the Caucasus populations and numerous populations of other geographic regions, considering 28 alleles of 12 loci of blood group, serum protein and red cell enzyme polymorphisms and constructed the dendrogram of these populations. The position of the Caucasus populations in the dendrogram corresponds on principle to the earlier anthropological observations. The clustering of the Caucasoid populations corresponds completely with anthropological and historical data, and supports our earlier hypothesis (Nazarova 1999) concerning the differentiation of Caucasoids, Northern Mongoloids and Amerinds from the populations, which inhabitated Asia in palaeolithic times.     

Kinematics and dynamics of the temporomandibular joint and of the movements of the mandible

In order to analyze the complicated movements of the mandible as the open-closing movement and the protrusio are, it is useful to evaluate the basic kinematic principles and reduce them to simple technical constructions. Both the open-closing movement and the protrusio could be reduced to 4-bar links, which were used to simulate the movements with help of a computer. Besides, the polodes and the curves of points in the muscular attachments could be constructed. The 2 entirely different 4-bar links have 3 things in common: The resting system - cranium, the moving system - mandibula, and 1 of the 2 arms connecting these 2 systems - the ligamentum laterale. As this ligament is taut during movements it can be considered a "guiding ligament" representing 1 of the 3 determining components of the mandibular movements. The other of the 2 arms has no anatomical equivalent; this arm, however, is "replaced" by the 2 other determining components of the mandibular movements: the joint and the muscles. The curves, which the Caput mandibulae describes, are practically identical for the open-closing movement and the protrusio despite of the different 4-bar links and these curves exactly correspond to the Discus articularis, taut by the upper part of the M. pterygoideus lateralis. The muscles do not only just move the mandibula, but they are also the component, which can choose between the different mandibular movements. By means of the curves, which points in the muscular attachments describe, the function of the masticatory muscles could be analyzed exactly.