Chemical investigation on a colonial marine tunicate, Didemnum sp. led to the isolation of a series of indole alkaloids including a new (1) and two known metabolites (2-3). Based on the spectroscopic analysis including 1D and 2D NMR along with MS spectra, the structure of 1 (16-epi-18-acetyl herdmanine D) was elucidated as a new amino acid derivative. The absolute configuration of 1 was determined by comparison of specific rotation with the known compound. The structures of compounds 2 and 3 were also identified as bromoindole containing compounds N-(6-bromo-1H-indole-3-carbonyl)-L-arginine and (6-bromo-1H-indol-3-yl) oxoacetamide, respectively, based on 1H and 13C NMR data, MS data and specific rotation value. Their pharmacological potentials as antibacterial agents and FXR antagonists were investigated, but no significant activity was found. However, the structural similarity of compound 1 to compound 4 suggested the antiinflammatory potential of compound 1.
Anti-Bacterial Agents ; Indole Alkaloids ; Urochordata*

BACKGROUND: In the last few decades, there has been a substantial increase in the population of aged people. Aging skin is a common concern for many people these days. OBJECTIVE: The purpose of this study was to evaluate the efficacy of cosmetics containing Ascidian tunic in reducing wrinkles. METHODS: Cream containing the Ascidian tunic was applied to right lateral epicanthal areas of 30 subjects for 8 weeks. As control group, a placebo which does not contain the Ascidian tunic was applied to the left lateral epicanthal areas of the subjects. Skin barrier function was evaluated and skin replicas were molded on both epicanthal areas every 4 weeks. The changes of the skin wrinkles were analyzed with skin visiometer. RESULTS: In comparison of the changes between groups treated with placebo and Ascidian tunic, there was statistically significant differences in hydration and sebum secretion. But there were no statistically significant differences in the result of R-value by visiometer. CONCLUSION: The treatment of Ascidian tunic extract seems to be partially effective in improving wrinkles, but further study is required.
Aged ; Aging ; Cosmetics ; Fungi ; Humans ; Sebum ; Skin ; Urochordata

Five purine and carboline alkaloids were isolated from the methanol extract of the ascidian Symplegma oceania. Classic chromatographies including preparative HPLC were used for isolation and purification of the compounds. The structures were established as 6-methoxy-7-methyl-8-oxoguanine (1), 2-methylimino-3-methyl-6-methylamino- 9H-purine (2), 1,2,3,4-tetrahydro-betacarboline (3), 1,2,3,4-tetrahydro-1-methyl-beta-carboline (4) and 1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acid (5) by comparison the spectroscopic data (MS, 1H, 13C-NMR) with those reported in the literatures. Compounds 2-5 were reported from the the genus Symplegma for the first time. The purine and carboline were the major alkaloid types of S. oceania.
Alkaloids ; chemistry ; Animals ; Carbolines ; chemistry ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Purines ; chemistry ; Urochordata ; chemistry

OBJECTIVES: To develop a bioactive membrane for guided bone regeneration (GBR), the biocompatibility and bone regenerating capacity of the cellulose membrane obtained from the Ascidians squirt skin were evaluated. MATARIALS AND METHODS: After processing the pure cellulose membrane from the squirt skin, the morphological study, amino acid analysis and the immunoreactivity of the cellulose membrane were tested. Total eighteen male Spraque-Dawley rats (12 weeks, weighing 250 to 300g) were divided into two control (n=8) and another two experimental groups (n=10). In the first experimental group (n=5), the cellulose membrane was applicated to the 8.0 mm sized calvarial bone defect and the same sized defect was left without cellulose membrane in the first control group (n=4). In the another experimental group (n=5), the cellulose membrane was applicated to the same sized calvarial bone defect after femoral bone graft and the same sized defect with bone graft was left without cellulose membrane in the another control group (n=4). Each group was sacrificed after 6 weeks, the histological study with HandE and Masson trichrome stain was done, and immunohistochemical stainings of angiogenin and VEGF were also carried out. RESULTS: The squirt skin cellulose showed the bio-inductive effect on the bone and mesenchymal tissues in the periosteum of rat calvarial bone. This phenomenon was found only in the inner surface of the cellulose membrane after 6 weeks contrast to the outer surface. Bone defect covered with the bioactive cellulose membrane showed significantly greater bone formation compared with control groups. Mesenchymal cells beneath the inner surface of the bioactive cellulose membrane were positive to the angiogenin and VEGF antibodies. CONCLUSION: We suppose that there still remains extremely little amount of peptide fragment derived from the basement membrane matrix proteins of squirt skin, which is a kind of anchoring protein composed of glycocalyx. This composition could prevent the adverse immunological hypersensitivity and also induce bioactive properties of cellulose membrane. These properties induced the effective angiogenesis with rapid osteogenesis beneath the inner surface of cellulose membrane, and so the possibilities of clinical application in dental field as a GBR material will be able to be suggested.
Animals ; Antibodies ; Basement Membrane ; Bone Regeneration* ; Cellulose* ; Glycocalyx ; Hand ; Humans ; Hypersensitivity ; Male ; Membranes* ; Osteoblasts ; Osteogenesis ; Periosteum ; Rats ; Skin* ; Transplants ; Urochordata ; Vascular Endothelial Growth Factor A

Ascidian soft tunic syndrome (AsSTS) caused by Azumiobodo hoyamushi (A. hoyamushi) is a serious aquaculture problem that results in mass mortality of ascidians. Accordingly, the early and accurate detection of A. hoyamushi would contribute substantially to disease management and prevention of transmission. Recently, the loop-mediated isothermal amplification (LAMP) method was adopted for clinical diagnosis of a range of infectious diseases. Here, the authors describe a rapid and efficient LAMP-based method targeting the 18S rDNA gene for detection of A. hoyamushi using ascidian DNA for the diagnosis of AsSTS. A. hoyamushi LAMP assay amplified the DNA of 0.01 parasites per reaction and detected A. hoyamushi in 10 ng of ascidian DNA. To validate A. hoyamushi 18S rDNA LAMP assays, AsSTS-suspected and non-diseased ascidians were examined by microscopy, PCR, and by using the LAMP assay. When PCR was used as a gold standard, the LAMP assay showed good agreement in terms of sensitivity, positive predictive value (PPV), and negative predictive value (NPV). In the present study, a LAMP assay based on directly heat-treated samples was found to be as efficient as DNA extraction using a commercial kit for detecting A. hoyamushi. Taken together, this study shows the devised A. hoyamushi LAMP assay could be used to diagnose AsSTS in a straightforward, sensitive, and specific manner, that it could be used for forecasting, surveillance, and quarantine of AsSTS.
Animals ; Euglenozoa Infections/diagnosis/veterinary ; Kinetoplastida/*classification/genetics/*isolation & purification ; Nucleic Acid Amplification Techniques/*methods ; Predictive Value of Tests ; RNA, Ribosomal, 18S/*genetics ; Sensitivity and Specificity ; Urochordata

This study was conducted to investigate food and meal preference of workers (435 male and 212 female) in the Chonnam Yeosu industrial area. The results of the survey were analyzed by principal components analysis. The results were obtained as follows: 19.3% of the subjects were twenties, 28.3% were thirties, 28.8% were forties and 23.7% were fifties. Females liked rice rolled in dried laver, rice cakes, janchi-noodle and breads, on the other hand males liked thick beef soup, loach soup, an eel stew and soju. The young liked instant noodles, fried chicken, sweet and sour pork, pork cutlet, pizza, hamburger, ham, sausage and fruit beverage, on the other hand the old liked a fish pot stew, loach soup, eel stew, fish boiled in soy with spices, panfried fish, sea slug, ascidian, bunder, green laver and boiled burdock-lotus root in soy. Females who are young liked hamburger and sweet and sour pork whereas males who are young liked instant noodles, pork cutlet, ham, sausage and fruit beverage. Aged females liked fish boiled in soy with spices, panfried fish, bunder, green laver, boiled burdock-lotus root in soy whereas aged males liked a fish pot stew, loach soup, eel stew, sea mussel and oysters. Boiled rice was located within the middle irrespective of age and sex, but noodles, wheat flour meal, fast foods and fruits were situated at the young female side. Aged males liked soup and pot stew. Young males liked meats and eggs whereas fish and shellfish and kimch were located at the aged people side. Aged females liked sea weeds and most people disliked vegetables but females liked some vegetables irrespective of age. Processed foods, salted foods, and alcohol were generally disliked foods by subjects but males liked those foods. Soybean curd was liked more males than females, and teas, except coffee, was liked by males.
Aged ; Beverages ; Bivalvia ; Bread ; Chickens ; Coffee ; Eels ; Eggs ; Fast Foods ; Female ; Flour ; Fruit ; Gastropoda ; Hand ; Humans ; Male ; Meals ; Meat ; Ostreidae ; Ovum ; Shellfish ; Soybeans ; Spices ; Tea ; Triticum ; Urochordata ; Vegetables

A new method based on high performance liquid chromatography-electrospray ionization time of flight-mass spectrometry (HPLC-ESI-TOF/MS) was developed for the rapid identification of active compounds in Styela clava and the development of its specific chromatograms. Samples were extracted by ultrasonic-assisted extraction, and the extraction conditions were optimized. The developed HPLC-ESI-TOF/MS method was used to identify the components in Styela clava extract, and a specific chromatogram based on HPLC analysis was established. Ten compounds in Styela clava extract have been primary identified by HPLC-ESI-TOF/MS on-line detection combined with literature review. The result of similarity evaluation for specific chromatograms indicated that the quality of different Styela clava samples was not entirely consistent. This method has the advantages of simple operation, rapid measurement and it is a powerful tool for identification of active components in Styela clava and its quality control.
Animals ; Chromatography, High Pressure Liquid ; methods ; Hypoxanthine ; analysis ; Quality Control ; Spectrometry, Mass, Electrospray Ionization ; methods ; Tyrosine ; analysis ; Uridine ; analysis ; Urochordata ; chemistry

Alkaloids ; chemistry ; isolation & purification ; pharmacology ; Animals ; Antineoplastic Agents ; pharmacology ; Antiviral Agents ; pharmacology ; Carbolines ; chemistry ; isolation & purification ; pharmacology ; Humans ; Molecular Structure ; Urochordata ; chemistry

Styela clava, called non-native tunicate or sea squirt, is habitat which include bays and harbors in Korea and several sites in the sea faced world. We fabricate cellulose membrane nerve conduit (CMNC) from this native sea squirt skin, and evaluate the capacity of promoting peripheral nerve regeneration in the rat sciatic nerve defect model. After processing the pure cellulose membrane from the sea squirt skin as we already published before, CMNC was designed as a non-tubular sheet with 14 mm length and 4 mm width. Total eleven male Spraque-Dawley rats (12 weeks, weighing 250 to 300g) were divided into sham group (n=2), silicone tube grafted control group (n=3) and experimental group (n=6). Each CMNC grafted nerve was evaluated after 4, 8 and 12 weeks in the experimental group, and after 12 weeks, sciatic function was evaluated with sciatic function index (SFI) and gait analysis, and histomorphology of nerve conduit and the innervated tissues of sciatic nerve were all examined using image analyzer and electromicroscopic methods in the all groups. The regenerated axon and nerve sheath were found only in the inner surface of the CMNC after 4 weeks and became more thicker after 8 and 12 weeks. In the TEM study, CMNC grafted group showed more abundant organized myelinated nerve fibers with thickened extracellular matrix than silicone conduit grafted group after 12 weeks. The sciatic function index (SFI) and ankle stance angle (ASA) in the functional evaluation were -47.2+/-3.9, 35.5.+/-4.9.in CMNC grafted group (n=2) and -80.4+/-7.4, 29.2.+/-5.3.in silicone conduit grafted group (n=3), respectively. And the myelinated axon was 41.59% in CMNC group and 9.51% in silicone conduit group to the sham group. The development of a bioactive CMNC to replace autogenous nerve grafts offers a potential and available approach to improved peripheral nerve regeneration. As we already published before, small peptide fragment derived from the basement membrane matrix proteins of squirt skin, which is a kind of anchoring protein composed of glycocalyx, induced the effective axonal regeneration with rapid growth of Schwann cells beneath the inner surface of CMNC. So the possibilities of clinical application as a peripheral nerve regeneration will be able to be suggested.
Animals ; Ankle ; Axons ; Basement Membrane ; Bays ; Cellulose* ; Ecosystem ; Extracellular Matrix ; Gait ; Glycocalyx ; Humans ; Korea ; Male ; Membranes* ; Myelin Sheath ; Nerve Fibers, Myelinated ; Peripheral Nerves* ; Rats ; Regeneration* ; Schwann Cells ; Sciatic Nerve ; Silicones ; Skin ; Transplants ; Urochordata

The ascidian Ciona intestinalis is a model organism of developmental and evolutionary biology and may provide crucial clues concerning two fundamental matters, namely, how chordates originated from the putative deuterostome ancestor and how advanced chordates originated from the simplest chordates. In this paper, a whole-life-span culture of C. intestinalis was conducted. Fed with the diet combination of dry Spirulina, egg yolk, Dicrateria sp., edible yeast and weaning diet for shrimp, C. intestinalis grew up to average 59 mm and matured after 60 d cultivation. This culture process could be repeated using the artificially cultured mature ascidians as material. When the fertilized eggs were maintained under 10, 15, 20, 25 degrees C, they hatched within 30 h, 22 h, 16 h and 12 h 50 min respectively experiencing cleavage, blastulation, gastrulation, neurulation, tailbud stage and tadpole stage. The tadpole larvae were characterized as typical but simplified chordates because of their dorsal nerve cord, notochord and primordial brain. After 8 - 24 h freely swimming, the tadpole larvae settled on the substrates and metamorphosized within 1- 2 d into filter feeding sessile juvenile ascidians. In addition, unfertilized eggs were successfully dechorionated in filtered seawater containing 1% Tripsin, 0.25% EDTA at pH of 10.5 within 40 min. After fertilization, the dechorionated eggs developed well and hatched at normal hatching rate. In conclusion, this paper presented feasible methodology for rearing the tadpole larvae of C. intestinalis into sexual maturity under controlled conditions and detailed observations on the embryogenesis of the laboratory cultured ascidians, which will facilitate developmental and genetic research using this model system.
Animals ; Ciona intestinalis ; growth & development ; Embryonic Development ; physiology ; Female ; Male ; Metamorphosis, Biological ; physiology ; Zygote ; growth & development