Internal Microanatomy and Zoological Affinity of the Early Cambrian Olivooides

The early Cambrian pentamerous microfossil Olivooides/Punctatus in South China, which is characterized by a diagnostic stellate tubular apex, has been well-known for its almost complete development sequence that can be confidently traced from embryos and hatched juveniles, to conical adults. However, its zoological affinity remains highly controversial. Here we describe the internal microanatomic structures of the soft-body inside the peridermal theca of Olivooides multisulcatus Qian, 1977, including interradial pairs of tentacles, adradial and perradial frenula, perradial oral marginal lappets and twins of perradial gastric saccule-like humps as well as a circular velarium with striated coronal muscles. Particularly, one specimen shows bifurcated velarial canals along the bell aperture. Both the components of the soft-body and the external theca （or cyst） are arranged in perfect pentaradial symmetry. These characteristics are more compatible with those of living cubomedusans and co-occurring Cambrian athecate embryonic cubozoans. Concerning the presence of peridermal theca, Olivooides most likely represents an extinct thecate stem-group cubomedusae but devoid of both perradial eyes and specialized pedalia. The well-grown soft body inside the peridermal tube displays a set of mixed features of both polyp and medusa.

Mitochondrial DNA control region diversity and population structure of Pacific herring(Clupea pallasii)in the Yellow Sea and the Sea of Japan

To investigate the genetic variation and population structure of Pacific herring in the Yellow Sea and the genetic differentiation between the Yellow Sea and the Sea of Japan, fragments of 479-bp mitochondrial DNA control region were sequenced for 110 individuals collected from three different periods in the Yellow Sea and one locality in the Sea of Japan. High haplotype diversity and moderate nucleotide diversity were observed in Pacific herring. AMOVA and exact test of population differentiation showed no significant genetic differentiations among the three populations of the Yellow Sea and suggested the populations can be treated as a single panmictic stock in the Yellow Sea. However, a large and significant genetic differentiation （ФST=0.11; P=0.00） was detected between the populations in the Yellow Sea and the Sea of Japan. The high sea water temperature in the Tsushima Strait was thought a barrier to block the gene exchange between populations of the two sea areas. The neutrality tests and mismatch distribution indicated recent population expansion in Pacific herring.

Continuous Saccharification of Laminarin by Immobilized Laminarinase ULam111 Followed by Ethanol Fermentation with a Marine-Derived Yeast

We isolated a novel laminarinase ULam111 from Flavobacterium sp. strain UMI-01. Purified ULam111 showed degradation activity against laminarin with the specific activity of 224 ± 18 U/mg at 30°C and pH 6.0. Its optimum temperature was 50°C, and degradation activities against laminarin were observed at 4°C - 80°C. With a laminarin degradation system, we investigated the preparation and properties of immobilized ULam111 with the use of the 11 types of carriers. The high activity recoveries of immobilized ULam111 were as follows: 19.4% for IB-S60P carrier beads (the non-ionic type), 15.6% for IB-S60S carrier beads (the non-ionic type), 11.9% for IB-150P carrier beads (the covalent type), and 7.1% for IB-C435 carrier beads (the cationic type). With the repeated use of immobilized ULam111, the enzyme activities immobilized on IB-S60S and those on IB-S60P remained at 40% and 30% respectively after the sixth trial. We selected IB-S60S as suitable beads for enzyme immobilization, and we attempted to construct a reactor system with ULam111 immobilized on IB-S60S beads. In this system, 1.2 - 1.9 g/L glucose was repeatedly produced from 30 mg/mL laminarin solutions after 20 hr when the reactor operation was repeated 10 times. We examined ethanol fermentation from the saccharified solutions with a marine-derived yeast (Saccharomyces cerevisiae C-19), and 0.51 - 0.58 g/L bioethanol was produced from the saccharified solution that contained 1.71 - 1.86 g/L of glucose.

Involvement of Cbl-b-mediated macrophage inactivation in insulin resistance

Aging and overnutrition cause obesity in rodents and humans. It is well-known that obesity causes various diseases by producing insulin resistance(IR). Macrophages infiltrate the adipose tissue(AT) of obese individuals and cause chronic low-level inflammation associated with IR. Macrophage infiltration is regulated by the chemokines that are released from hypertrophied adipocytes and the immune cells in AT. Saturated fatty acids are recognized by toll-like receptor 4(TLR4) and induce inflammatory responses in AT macrophages(ATMs). The inflammatory cytokines that are released from activated ATMs promote IR in peripheral organs, such as the liver, skeletal muscle and AT. Therefore, ATM activation is a therapeutic target for IR in obesity. The ubiquitin ligase Casitas b-lineage lymphoma-b(Cbl-b) appears to potently suppress macrophage migration and activation. Cbl-b is highly expressed in leukocytes and negatively regulates signals associated with migration and activation. Cbl-b deficiency enhances ATM accumulation and IR in aging- and diet-induced obese mice. Cbl-b inhibits migration-related signals and SFA-induced TLR4 signaling in ATMs. Thus, targeting Cbl-b may be a potential therapeutic strategy to reduce the IR induced by ATM activation. In this review, we summarize the regulatory functions of Cbl-b in ATMs.

Vitamin B6 and Lipid Contents in Engraulisjaponica Specifically Caught for Production of Japanese Soup Stock

The purpose of this study was to elucidate the relationship between percentage of body lipid and individual vitamin B6 contents in the anchovy used for production of Japanese soup stock. The contents of individual vitamin B6 compound in foods should be determined to estimate the functionality of foods. Anchovies （Engraulisjaponica） for use as production of iriko, which is a raw material for preparation of Japanese soup stock, were caught using a medium-sized purse seine on May 26th, June 1st, and July 6th 2011 in Tachibana Bay, Nagasaki, Japan. Vitamin B6 contents were analyzed by 4-pyridoxolacone-coversion （all-enzymatic） HPLC. The relationship between body length, lipid content and the individual contents of vitamin B6 compounds in the anchovies was determined. The propriety of anchovies for iriko preparation could not be estimated based on the appearance of length of fishes. The anchovies were rich in vitamin B6, especially pyridoxamine 5＇-phosphate, which may prevent diabetic complications. The percentage of body lipid significantly showed a negative correlation with the content of pyridoxamine 5＇-phosphate. The contents of the other vitamin B6 compounds did not show the significant correlation with the percentage of body lipid.

Molecular Probes in Tandem Electrospray Ionization Mass Spectrometry: Application to Tracing Chemical Changes of Specific Phospholipid Molecular Species

New ionization and detection techniques in mass spectrometry have been successfully applied for efficient analyses of complex biological systems. It is, however, still difficult to trace structural changes of a specific molecular species in such systems. In the present study, a molecular probe strategy in combination with tandem electrospray ionization mass spectrometry has been examined using synthetic deuterium-labeled phosphatidylcholine hydroperoxide (PC-OOH/D3) and ethyl-labeled phosphatidylcholine having docosahexaenoic acid side chain (DHA-PC/Et). Administration of a mixture of PC-OOH/D3 and DHA-PC/Et to human blood and human skin surface, followed by extraction and analysis with collision-induced tandem electrospray ionization mass spectrometry demonstrated that metabolites of both molecular probes can be detected simultaneously with strict selectivity. The present method is also found to be useful in tracing chemical changes of the unstable docosahexaenoyl group on the surface of processed fish. The activity of phospholipase A2 can also be assessed using a phospholipid molecular probe with a linoleoyl and a deuteriomethyl group via selective detection of the lyso-phospholipid product by mass spectrometry. The advantage of the present method is that no chromatographic separation is required and analysis can be performed under strictly the same condition for different molecular probes, affording multiple data by one experiment. The present strategy may be useful for tracing time-dependent phenomena in dynamic phospholipid biochemistry, and can be widely used for any biological and food systems.

Recent Advances in Researches on Physiologically Active Substances in Holothurians

In this report, we reviewed recent literature on physiologically active substances from sea cucumbers (SCs) andtheir activities together with results obtained from our study. Preventive properties against lipid metabolism were reported inrats using a whole SC preparation with no particular constituent specified. Administration of the preparation lowered serumand hepatic cholesterol levels and improved the HDL/LDL ratio. These functions may be attributed to the stimulatory effectof the extract on the secretion of cholesterol in feces. Novel fucosylated chondroitin sulfates (FCSs) from Ludwigothureagrisea significantly induced fibroblast growth factor 2-dependent angiogenesis in human umbilical vein endothelial cells (HU-VECs). The proangiogenetic activity seemed attributable to the action of the sulfated fucose branches on the polysaccharide.SCs contain mycosporine-like amino acids (MAAs) that are capable of absorbing UV. A biogenetic precursor of MAAs wasfirst reported in SCs. The anti-proliferative effects of a branched chain fatty acid from a sea cucumber on prostate cancercells was reported with the activity of 5-lipoxygenase. Glycosphingolipid constituents in SCs have been systematically ana-lyzed over the past ten years. The results showed that the gangliosides in several SCs differed from those of mammals in thata sialic acid of SC gangliosides directly binded to glucose of cerebroside. Neuritogenic activity of the glycosphingolipids wasdemonstrated in vitro experiments and may lead to the development of therapeutic products for neurological disorders. Ourstudy also showed that sphingoid bases, the hydrolyzed products of glycosphingolipids from SCs, induced significant apoptosisin several tumor cell lines.

Difference in Nitrogen Starvation-Inducible Expression Patterns among Phylogenetically Diverse Ammonium Transporter Genes in the Red Seaweed <i>Pyropia yezoensis</i>

Nitrogen deficiency induces senescence and the expression of genes encoding ammonium transporters (AMTs) in terrestrial plants where the AMT family is subdivided into AMT1 and AMT2 subfamilies. Nitrogen starvation in the red seaweed Pyropia yezoensis causes senescence-like discoloration. In this study, we identified five genes in P. yezoensis encoding AMT domain-containing proteins, which were phylogenetically categorized into the AMT1 subfamily. We also found a gene encoding a Rhesus protein (Rh) that was related to, but diverged from, AMTs. Moreover, our phylogenetic analysis showed that AMT domain-containing proteins from micro- and macro-algae belonged to either the AMT1 or Rh subfamily, indicating the absence of AMT2 in algae. Gene expression analyses revealed the presence of gametophyte- and sporophyte-specific AMT1 genes that were up-regulated transiently and continually, respectively, under nitrogen-deficient conditions. In addition, up-regulated sporophyte-specific gene expression was suppressed when nitrogen was resupplied. Accordingly, an expansion of the ancient AMT gene has produced AMT1 functional variants differing in temporal and nitrogen starvation-inducible expression patterns during the life cycle of P. yezoensis. These findings help elucidate the unique nutrition starvation responses involving functionally diverse AMT1 and Rh subfamilies in red seaweed.

Evolutionary relationship of three mitten crabs(Eriocheir sp)revealed by mitogenome and 5S ribosomal DNA analysis

To reveal the evolutionary relationship of three mitten crabs(Eriocheir sinensis,E.hepuensis,and E.japonica),complete mitogenomes and nuclear 5S rDNA sequences were analyzed.Sequencing revealed that the mitogenomes analyzed shared conserved organization of the coding and non-coding regions but genetic variation was identified.Among the three mitten crabs distinct tandem repeats were identified in the mitochondrial D-loop region.The 5S gene(5S rDNA)sequence was highly conserved across the three species,whereas non-transcribed spacer(NTS)region exhibit high levels of variation including insertions,deletions and point mutations.Cluster analysis suggested that the three mitten crabs had their own independent 5S rDNA sequence variation and evolutionary pattern.Both mitogenome and 5S rDNA sequence analysis revealed significant genetic variation across the mitten crabs species.Phylogenetic analysis using mitogenome and 5S rDNA sequences demonstrated that E.japonica was relatively more distant from E.sinensis and E.hepuensis.This study extended our previous knowledge and confirmed that the three mitten crabs are likely to be genetically differentiated species.In addition,our study also provided insights into the conservation of pure natural resources of E.sinensis,an important aquaculture species.