Tichocarpus Crinitus Research Articles

Comparative study of HIV-1 inhibition efficiency by carrageenans from red seaweeds family gigartinaceae, Tichocarpaceae and Phyllophoraceae

The efficiency of human immunodeficiency virus-1 (HIV-1) inhibition by sulfated polysaccharides isolated from the various families of red algae of the Far East Pacific coast were studied. The anti-HIV-1 activity of kappa and lambda-carrageenans from Chondrus armatus, original highly sulfated X-carrageenan with low content of 3,6-anhydrogalactose from Tichocarpus crinitus and i/κ–carrageenan with hybrid structure isolated from Ahnfeltiopsis flabelliformis was found. The antiviral action of these polysaccharides and its low-weight oligosaccharide was compared with commercial κ-carrageenan. Here we used the HIV-1-based lentiviral particles and evaluated that these carrageenans in non-toxic concentrations significantly suppress the transduction potential of lentiviral particles pseudotyped with different envelope proteins, targeting cells of neuronal or T-cell origin. The antiviral action of these carrageenans was confirmed using the chimeric replication competent Mo-MuLV (Moloney murine leukemia retrovirus) encoding marker eGFP protein. We found that X-carrageenans from T. crinitus and its low weight derivative and λ-carrageenan from C. armatus effectively suppress the infection caused by retrovirus. The obtained data suggest that the differences in the suppressive effect of carrageenans on the transduction efficiency of HIV-1 based lentiviral particles may be related to the structural features of the studied polysaccharides.

Algicella marina gen. nov., sp. nov., a novel marine bacterium isolated from a Pacific red alga.

A novel Gram-staining negative, strictly aerobic, rod-shaped, and non-motile bacterium, designated strain 9Alg 56T, was isolated from the red alga Tichocarpus crinitus. The phylogenetic analysis based on 16S rRNA gene sequences placed the novel strain within the family Rhodobacteraceae, the order Rhodobacterales, the class Alphaproteobacteria, the phylum Pseudomonadota. The nearest neighbors of the new strain were Pontivivens insulae KCTC 42458T, Oceanibium sediminis KCTC 62076T, Halovulum dunhuangense YYQ-30T and Monaibacterium marinum C7T with 16S rRNA gene sequence similarity of 94.7, 94.4%, 93.1 and 92.7%, respectively. The AAI/ANI/dDDH values between 9Alg 56T and the five species of the closest genera (Pontivivens, Oceanibium, Halovulum, Monaibacterium, and 'Oceanomicrobium') were 58.63-63.91%/ 75.91-77.37%/ 19.3-20.4%. The prevalent fatty acids of strain 9Alg 56T were C18:1 ω7c, C18:0 and C14:0 3-OH. The polar lipid profile consisted of phosphatidylethanolamine, phosphatidylglycerol, phosphatidic acid, phosphatidylcholine, and two unidentified lipids. The DNA G+C content of strain 9Alg 56T was 61.5mol%. A combination of the genotypic and phenotypic data showed that the algal isolate represents a novel genus and species, for which the name Algicella marina gen. nov., sp. nov. is proposed. The type strain is 9Alg 56T (= KCTC 72005T = KMM 6775T).

New Insights into the Structure of Kappa/Beta-Carrageenan: A Novel Potential Inhibitor of HIV-1.

New insights into the structure of the hybrid κ/β-carrageenan (κ/β-CRG) of the red alga Tichocarpus crinitus have been obtained. Carrageenan oligosaccharides were prepared through the chemical and enzymatic depolymerization of κ/β-CRG with κ-carrageenase and its the enzyme-resistant fraction. The composition and distribution of the repetition units of κ/β- CRG were investigated by using the negative ion tandem MALDI-TOFMS and ESIMS method, which made it possible to prove and characterize the hybrid structure of this polysaccharide. An analysis revealed the blockwise distribution of the long β-blocks along the polysaccharide chain, with the inclusion of κ/β, μ/ν-blocks and some ι-blocks. Furthermore, the desulfated κ/β-CRG was shown to contain of –G–D– repeating units up to 3.5 kDa. Previous studies have demonstrated that CRGs suppress the replication of several viruses. Here, we established that κ/β-CRG and its oligosaccharides significantly inhibit the transduction efficiency of replication-defective lentiviral particles pseudotyped with the envelope proteins of three different viruses. We found that the polysaccharide and its oligosaccharides strongly reduced the transduction efficiency of lentiviral particles pseudotyped with GP160—the envelope protein of the human immunodeficiency virus HIV-1—when added to T-lymphocyte Jurkat cells. The CRG oligosaccharides displayed significantly higher antiviral activity.

Morphological characterization of biominerals from five multicellular marine algae species

Silica biominerals are deposited as amorphous solid structures in plant cells and tissues, providing rigidity to different plant parts and assisting in defence. The shape and size of phytoliths are well established and serve as a useful tool in taxonomic analyses. For the first time we extracted and studied silica biominerals of five marine macroalgae, which we observed by light microscopy, scanning electron microscopy, and X-ray diffraction analysis (XRD). More than nine different morphotypes of phytoliths ranging from ≥ 10 to ≥ 350 μm in size were found. Some of them were phytoliths made of silica while others showed characteristics of different minerals of calcium. In our study, the “honeycomb” formations were only recorded in Laurencia tropica Yamada and pyramid tabular ones were found only in Tichocarpus crinitus (S.G. Gmelin) Ruprecht. The XRD analysis showed that they consisted of virgilite and gypsum substance, respectively. Silica phytoliths are intrinsic parts of the algae and their morphological characterization can provide the basis for palaeo-reconstruction and taxonomic investigation of brown and red algae in palaeontological studies of fossils where all organic matter has decayed.

Hexagonal Phytolithes from Red Alga <i>Tichocarpus crinitus</i>

In current work unique hexagonal microparticles of silicon oxide (phytolite), isolated from the red algae Tichocarpus crinitus, which grows in the Sea of Japan described. The structures of poytolites are characterized by the methods of optical microscopy and scanning electron microscopy. The role of these formations in plant life and the mechanism of their synthesis remains unclear.

MORPHO-FUNCTIONAL CHARACTERISTICS OF GAMETES AND PRODUCING POSSIBILITIES OF MARINE ORGANISMS FOR THEIR REPRODUCTION IN MONO- AND POLYCULTURE

Reproduction of the sea urchins Strongylocentrotus intermedius, Mesocentrotus nudus, and the scallop Mizuhopecten yessoensis is considered in natural communities with various algae: red (Gracilaria verrucusa and Tichocarpus crinitus), brown (Sacharina cichorioides), and green (Vulva fenistrata). In these environments, their reproduction is influenced by exometabolites released by macrophytes, so the gametes with different production capacity could be formed and products of metabolism could impede their growth and development. To avoid these negative effects in polyculture, the cultured community should include species of different trophic levels forming a single food chain in which their exometabolites are utilized. The polyculture could be controlled by changing the number of species or influence on them by change of abiotic factors. In artificial polyculture, optimal conditions for reproduction and growth of cultivated species could be created, that provides the maximum production from a limited area.

Aquimarina algiphila sp. nov., a chitin degrading bacterium isolated from the red alga Tichocarpus crinitus.

A strictly aerobic, Gram-stain-negative, rod-shaped, motile by gliding and yellow-orange pigmented flavobacterium, designated strain 9Alg 151T, was isolated from the Pacific red alga Tichocarpus crinitus. Phylogenetic analysis based on 16S rRNA gene sequences showed that the novel strain fell into the genus Aquimarina of the family Flavobacteriaceae with a 16S rRNA gene sequence similarity range of 94.2-98.2 % to the recognized species of the genus. Strain 9Alg 151T grew in the presence of 0.5-5 % NaCl and at 5-34 °C, and hydrolysed aesculin, agar, gelatin, starch, Tween 40, DNA and chitin. The predominant fatty acids were iso-C17 : 0 3-OH, iso-C15 : 0, iso-C15 : 1 G, iso-C15 : 0 3-OH, iso-C16 : 0, iso-C17 : 1ω8c and summed feature 3. The polar lipid profile comprised phosphatidylethanolamine, three unidentified aminolipids and three unidentified lipids. The major respiratory quinone was MK-6. The genomic DNA G+C content was 32.6 mol%. On the basis of 16S rRNA gene sequence data, and chemotaxonomic and phenotypic characteristics, strain 9Alg 151T represents a novel species of the genus Aquimarina, for which the name Aquimarina algiphila sp. nov. is proposed. The type strain is 9Alg 151T (=KCTC 23622T=KMM 6462T).

Carrageenanolytic enzymes from marine bacteria associated with the red alga Tichocarpus crinitus

One hundred six bacterial strains (106 strains) were isolated from the Pacific red alga Tichocarpus crinitus, collected from the Troitsa Bay of Gulf of Peter the Great (Sea of Japan, Russia), and studied on their ability to degrade different samples of carrageenan as substrates. Some of the studied strains exhibited hydrolytic activity selectively to a total polysaccharide (kappa and lambda) from Chondrus armatus, other strains, to κ/β-carrageenan from T. crinitus, which did not possess the same effect to a total polysaccharide from T. crinitus. The strains with the highest enzyme activity (28 strains) were identified using 16S rRNA gene sequence techniques and classified to the phyla Bacteroidetes and Proteobacteria. Most (75%) of the studied carrageenase producers belong to phylum Bacteroidetes (21 strains). These strains were related with 7 phylotypes of the genera Aquimarina, Cellulophaga, Maribacter, and Zobellia (family Flavobacteriaceae of the class Flavobacteriia). Cellulophaga strains were the dominant group (35.7% of total) and closely related to species C. baltica and C. lytica with 99.8 and 99.9% sequence similarity, respectively. The remaining strains (7 strains) were represented by members of the genera Altererythrobacter (the family Erythrobacteraceae), Phaeobacter, Sulfitobacter, Tateyamaria (the family Rhodobacteraceae), and Sphingomonas (the family Sphingobacteriaceae) of the class Alphaproteobacteria and Psychrobacter (the family Moraxellaceae) and Vibrio (the family Vibrionaceae) of the class Gammaproteobacteria.

Influence of carrageenan on cytokine production and cellular activity of mouse peritoneal macrophages and its effect on experimental endotoxemia.

The in vivo effect of κ/β-carrageenan isolated from the red alga Tichocarpus crinitus on cytokine synthesis and cellular activity of murine peritoneal macrophages and also the protective effect of polysaccharides in LPS-induced endotoxemia in mice was studied. It was established that κ/β-carrageenan given orally at a dose of 100 mg/kg stimulates the induction of anti-inflammatory cytokines (IL-10) in mouse blood cells by more than 2.5-fold compared with control, with no effect on pro-inflammatory cytokine (TNF-α) production. Pretreating mice with carrageenan once a day before injecting LPS increased the levels of IL-10 by 2.5-fold and reduced TNF-α production by 2-fold compared with control. So, κ/β-carrageenan alone and in combination with LPS enhanced the cellular activity and mobility of peritoneal macrophages by increasing cell adhesion and migration compared with control. LPS activated cells intensively, sometimes resulting in their destruction by necrosis; carrageenan pretreatment reduced the excessive inflammatory cell activation caused by LPS. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1549-1557, 2017.

Examination of the structures of several glycerolipids from marine macroalgae by NMR and GC-MS.

Several classes of glycerolipids were isolated from the total lipids of the algae Saccharina cichorioides, Eualaria fistulosa, Fucus evanescens, Sargassum pallidum, Silvetia babingtonii (Ochrophyta, Phaeophyceae), Tichocarpus crinitus, and Neorhodomela larix (Rhodophyta, Florideophyceae). The structures of these lipids were examined by nuclear magnetic resonance (NMR) spectroscopy, including 1D ((1) H and (13) C) and 2D (COSY, HSQC and HMBC) experiments. All of the investigated algae included common galactolipids and sulfonoglycolipids as the major glycolipids. Minor glycolipids isolated from S. cichorioides, T. crinitus, and N. laris were identified as lyso-galactolipids with a polar group consisted of the galactose. Comparison of the (1) H NMR data of minor nonpolar lipids isolated from the extracts of the brown algae S. pallidum and F. evanescens with the (1) H NMR data of other lipids allowed them to be identified as diacylglycerols. The structures of betaine lipids isolated from brown algae were confirmed by NMR for the first time. The fatty acid compositions of the isolated lipids were determined by gas chromatography-mass spectrometry.

Oligosaccharides of κ/β-carrageenan from the red alga Tichocarpus crinitus and their ability to induce interleukin 10

The structural peculiarities of oligosaccharide mixture (LMW-carr) obtained by mild acid hydrolysis of κ/β-carrageenan from the red alga Tichocarpus crinitus were investigated by negative-ion MALDI-TOFMS. An analysis suggested the presence of β-, ι- and γ-carrageenan (the precursor of β-carrageenan) blocks. MS/MS analysis revealed the presence of β/ι-hybrid structures, which co-existed in the hydrolysis mixture along with κ/β-tetrasaccharide. The effect of source polysaccharide (400 kDa) and purified oligosaccharides LMW-carr (1.7 kDa) on the production of anti-inflammatory cytokine IL-10 was examined in experiments in vivo and ex vivo. Both samples of carrageenan stimulated induction of IL-10 in human and in mice blood cells that depended on their molecular weights. LMW-carr (administered orally) induced more IL-10 in vivo production than source polysaccharide. The ex vivo cytokine-induced activity of HMW-carr did not depend on its concentration. LMW-carr caused strong increase in the level of cytokines at high concentration (10 μg mL−1).

Distribution of sublittoral vegetation at the continental coast of the Okhotsk Sea (within the limits of Khabarovsk Region)

Spatial distribution of algae vegetation is considered for the area from the Uda Bay to the Ushki Bay (1100 km along the northwestern coast of the Okhotsk Sea) on the data of 10 surveys conducted by Pacific Fish. Res. Center (TINRO) at the depth from 2 to 12-35 m in 2002-2013 (3043 stations, including about 300 diving stations made by the author personally). Dominant species of sublittoral vegetation are few: 4 species of brown algae ( Saccharina gurjanovae, Alaria marginata, Pseudolessonia laminarioides , and Stephanocystis crassipes ) and a group of red algae ( Neoptilota asplenioides, Odonthalia corymbifera, Tichocarpus crinitus, Chondrus platynus, Neohypophyllum middendorffii , and Hymenena ruthenica ), other species can form the vegetation cover in local areas only. The algae belt reaches the depth 8-13 m, sometimes 26-35 m. Its vegetation is qualitatively homogeneous, with feebly marked vertical zoning and tier structure; the dominant species don’t change by latitude and topical conditions. Mean biomass of the sublittoral algae is about 2 kg/m 2 , with the greatest contribution of Saccharina gurjanovae (annual mean 41 %) and the minimal one of Pseudolessonia laminarioides (13 %). The species composition is relatively stable in time, as well. The features of vegetation homogeneity are shown in over 900 km of the coastal zone. However, the sublittoral vegetation changes significantly southwestward and northeastward from the investigated area.

Effects of water soluble extracts from marine algae on root growth in soybean seedlings

Data on the effects of water soluble extracts of red marine algae—such as Grateloupia divaricate, Chondrus pinnulatus, Neorhodomela larix, and Tichocarpus crinitus—brown marine algae—such as Stephanocystis crassipes, Coccophora langsdorfii, Sphaerotrichia divaricata, Saccharina japonica, Sargassum pallidum, and Chorda filum—and green marine algae—such as Ulva fenestrata and Codium fragile—on root growth in soybean seedlings (Glycine max (L.) Merr.) have been presented. The stimulatory effects of extracts containing T. crinitus, N. Larix, C. pinnulatus, and S. crassipes algae at low (10−4 g/mL) and ultralow (10−10–10−12 g/mL) concentrations are defined.

Polysaccharide structure of tetrasporic red seaweed Tichocarpus crinitus

Sulfated polysaccharide isolated from tetrasporic plants of Tichocarpus crinitus was investigated. The polysaccharide was isolated by two methods: with water extraction at 80°C (HT) and with a mild alkaline extraction (AE). The extracted polysaccharides were presented by non-gelling ones only, while galactose and 3,6-AG were the main monosaccharides, at the same time amount of 3,6-AG in AE polysaccharides was the similar to that of HT. According to methods of spectroscopy and mass spectrometry, the polysaccharide from tetrasporic T. crinitus contains main blocks of 1,3-linked β-d-galactopyranosyl-2,4-disulfates and 1,4-linked 3,6-anhydro-α-d-galactopyranosyl while 6-sulfated 4-linked galactopyranosyl resudies are randomly distributed along the polysaccharide chain. The alkaline treatment of HT polysaccharide results in obtaining polysaccharide with regular structure that composed of alternating 1,3-linked β-d-galactopyranosyl-2,4-disulfates and 1,4-linked 3,6-anhydro-α-d-galactopyranosyl residues. Native polysaccharide (HT) possessed both high anticoagulant and antiplatelet activity measured by fibrin clotting and platelet aggregation induced by collagen. This activity could be connected with peculiar chemical structure of HT polysaccharide which has high sulfation degree and contains also 3,6-anhydrogalactose in the polymer chain.

Low molecular weight derivatives of different carrageenan types and their antiviral activity

A number of low molecular weight (LMW) fractions of carrageenans with different structural types were obtained by free radical depolymerization (H2O2), mild acid hydrolysis (HCl), and a specific enzyme. Three samples of carrageenans were depolymerized: kappa-carrageenan from Chondrus armatus, kappa-carrageenan from Kappaphycus alvarezii, and kappa/beta-carrageenan from Tichocarpus crinitus with initial molecular weights of 250, 390, and 400 kDa, respectively. The chemical depolymerization by two methods resulted to LMW derivatives of carrageenans with molecular weight from 1.2 to 3.5 kDa. Oligosaccharides of kappa- and kappa/beta-carrageenans with molecular weight of 2.2 and 4.3 kDa, respectively, were obtained after enzymatic depolymerization by recombinant kappa-carrageenase from Pseudoalteromonas carrageenovora. It was shown that the antiviral activity of high molecular weight carrageenans against tobacco mosaic virus was higher than that of their LWM derivatives independently on the depolymerization method. The method of depolymerization had some influence on the antiviral activity of carrageenan. LMW derivatives of kappa- and kappa/beta-carrageenans obtained by mild acid hydrolysis showed higher antiviral activity than the products of free radical depolymerization. The oligosaccharides prepared by enzymatic degradation possessed the lowest activity.

Comparison of the structures of hybrid κ-/β-carrageenans extracted from Furcellaria lumbricalis and Tichocarpus crinitus

Abstract κ-/β-Carrageenans extracted from Furcellaria lumbricalis and Tichocarpus crinitus are hybrid polysaccharides that have roughly similar κ- and β-carrabiose contents. The distributions of these carrabiose moieties were compared using gel permeation chromatography to analyse the end-products obtained after degradation with Pseudoalteromonas carrageenovora κ-carrageenase. Three fractions were obtained: standard oligo-κ-carrageenans, hybrid oligo-κ-/β-carrageenans and an enzyme-resistant fraction. The structure of the most abundant hybrid oligo-κ-/β-carrageenans and the composition of the κ-/β-fractions resistant to degradation were determined by 1H NMR. Altogether, the results suggest that the κ- and β-carrabiose moieties in F. lumbricalis are more randomly distributed than in T. crinitus carrageenan, which has a more regular block-type distribution.

Seasonal dynamics of subtidal macrophyte assemblages in Sobol Bay (Peter the Great Bay, Sea of Japan) in relation to depth

This paper describes the species composition, vertical distribution, and the seasonal changes in the biomass of subtidal macrophytes in Sobol Bay (Peter the Great Bay, Sea of Japan). The samples were collected in the depth-range from 0.5 to 4 m. A total of 65 macrophyte species were identified, among which were 10 species of Chlorophyta, 17 of Phaeophyceae, 37 of Rhodophyta and 1 of Magnoliophyta. Multivariate analyses were performed to detect spatial and temporal variations. Maximum species richness was registered in June, with a particularly dramatic biomass increase of brown algae. Both the number of species and the biomass of macrophytes decreased with increasing depth. Depth clearly affected the patterns of seasonal fluctuations of the species composition and biomass. In the study area, the species composition of the shallow-water algal assemblages was more stable throughout the year compared to that of the algal assemblages found in deeper waters. In total, four macrophyte assemblages were identified in the bay. A Phyllospadix iwatensis and Coccophora langsdorfii dominated assemblage occupied muddy-gravel bottoms in depths from 0.5 to 2 m throughout the year. An assemblage, co-dominated by annual brown algae Desmarestia viridis and Costaria costata, occurred at depths more than 3 m at late spring. Two other assemblages occurred on the rock and boulder bottom at 0.5–2 m depths, but they were temporally separated. An assemblage co-dominated by annual laminarian algae (Undaria pinnatifida and Costaria costata) developed at late spring and was succeeded in the autumn and winter by a Tichocarpus crinitus, Ulva lactuca, Sargassum pallidum and Codium fragile co-dominated assemblage. A comparison between the data obtained in the present study and the results of a previous study conducted in 1927 shows that the macrophytes assemblages in Sobol Bay underwent little change.

Analysis of structural heterogeneity of κ/β-carrageenan oligosaccharides from Tichocarpus crinitus by negative-ion ESI and tandem MALDI mass spectrometry

A mixture of oligosaccharides, released with enzymatic digestion from κ/β-carrageenan of the red seaweed Tichocarpus crinitus was investigated. The composition, sequence and sulfation pattern of the oligosaccharide mixture were elucidated by negative-ion electrospray ionization and MALDI potential lift tandem TOF mass spectrometry. It was shown, that oligosaccharide mixture (DP up to 20), produced by κ-carrageenase, contained κ-carrabiose, κ-carratetraose as main components, and set of hybrid oligosaccharides as less abundant components: β→κ, κ→β→κ, β→κ→κ, and minor β→ι→κ, β→β→κ→κ, β→κ→β→κ, κ→β→β→κ, κ→β→κ→κ κ→β→β→β→κ, β→β→κ→β→κ, κ→β→κ→β→κ, κ→β→β→κ→κ. The application of arabinoosazone as a MALDI matrix for negative-ion mode with the addition of l-fucose allowed decreasing the in-source fragmentation. The oligosaccharides of κ/β-carrageenan were shown to maintain an ability to induce of TNF-α and IL-6 secretion of immune cells.

Inhibitory effect of κ/β-carrageenan from red alga Tichocarpus crinitus on the development of a potato virus X infection in leaves of Datura stramonium L.

The effect of kappa/beta-carrageenan from red alda Tichocarpus crinitus on the development of a potato virus X (PVX) infection in the leaves of Datura stramonium L. has been studied. The treatment of leaves with carrageenan stimulates a protein synthesis in the cells, causing an increase in the size of nucleoli and in the number of mitochondria and membranes of the granular endoplasmic reticulum. At the same time, such treatment slightly stimulates lytic processes, causing an increase in the number of agranular endoplasmic reticulum cisterns, dictyosomes, and cytoplasmic vacuoles and the formation of cytoplasmic zones, transparent to electron microscopy. The carrageenan-induced stimulation of lytic processes results in the destruction of viral particles and can be considered as one of the defense mechanisms, preventing the intracellular accumulation of viruses. The carrageenan-stimulated formation of PVX-specific laminar structures, able to bind viral particles and, therefore, prevent their intracellular translocation and reproduction, represents another carrageenan-induced mechanism of the antiviral defense in plant cells.

Characteristics of polysaccharides and protein associated with them from dried and freshly collected red alga Tichocarpus crinitus

Polysaccharides isolated by successive extraction with water at 20 and 80°C from freshly collected and dried alga T. crinitus were compared. It was shown that the yield of polysaccharides from freshly collected alga was 40-44%; from dried material, less than 25%. It was found that the amount of extracted polysaccharides and their molecular weights decreased upon storage of dried alga for three years. Polysaccharides isolated from freshly collected and dried alga had identical structures and were a mixture of κ/β- and a new X-type of carrageenan. It was shown that protein, the amount of which reached 24% in the extracts obtained at 20°C, was strongly bound to the carrageenan. The amino-acid compositions of the proteins associated with the polysaccharides isolated at 20 and 80°C were identical and had an elevated content of serine, glutamic acid, glycine, and alanine.