NITROGEN FIXATION IN BACTERIA AND HIGHER PLANTS
CONTENTS COMMERCIAL FUCOIDANS FROM FUCUS VESICULOSUS CAN BE GROUPED INTO ANTIADIPOGENIC AND ADIPOGENIC AGENTS 1 ANTI OBESITY EFFECT OF CHITOSAN OLIGOSACCHARIDE CAPSULES COSCS IN OBESE RATS BY AMELIORATING LEPTIN RESISTANCE AND ADIPOGENESIS 15 ANTI OBESITY EFFECTS OF COLLAGEN PEPTIDE DERIVED FROM SKATE RAJA KENOJEI SKIN THROUGH REGULATION OF LIPID METABOLISM 31 CHITOSAN OLIGOSACCHARIDES IMPROVE GLUCOLIPID METABOLISM DISORDER IN LIVER BY SUPPRESSION OF OBESITY RELATED INFLAMMATION AND RESTORATION OF PEROXISOME PROLIFERATOR ACTIVATED RECEPTOR GAMMA PPAR 43 REDUCED NUMBER OF ADIPOSE LINEAGE AND ENDOTHELIAL CELLS IN EPIDIDYMAL FAT IN RESPONSE TO OMEGA 3 PUFA IN MICE FED HIGH FAT DIET 58 INHIBITION OF ADIPOGENESIS BY DIPHLORETHOHYDROXYCARMALOL DPHC THROUGH AMPK ACTIVATION IN ADIPOCYTES 76 EFFECT OF ORAL INGESTION OF LOW MOLECULAR COLLAGEN PEPTIDES DERIVED FROM SKATE RAJA KENOJEI SKIN ON BODY FAT IN OVERWEIGHT ADULTS A RANDOMIZED DOUBLE BLIND PLACEBO CONTROLLED TRIAL 85 ANTI OBESITY AND ANTI DIABETIC EFFECTS OF ISHIGE OKAMURAE 97 CHLOROPHYLL DERIVATIVES FROM MARINE CYANOBACTERIA WITH LIPID REDUCING ACTIVITIES 108 HYPOLIPIDEMIC EFFECT OF ARTHROSPIRA SPIRULINA MAXIMA SUPPLEMENTATION AND A SYSTEMATIC PHYSICAL EXERCISE PROGRAM IN OVERWEIGHT AND OBESE MEN A DOUBLE BLIND RANDOMIZED AND CROSSOVER CONTROLLED TRIAL 126 IDENTIFICATION OF CYANOBACTERIAL STRAINS WITH POTENTIAL FOR THE TREATMENT OF OBESITY RELATED CO MORBIDITIES BY BIOACTIVITY TOXICITY EVALUATION AND METABOLITE PROFILING 139 APPLICATION OF BIOACTIVE THERMAL PROTEOME PROFILING TO DECIPHER THE MECHANISM OF ACTION OF THE LIPID LOWERING 132 HYDROXY PHEOPHYTIN ISOLATED FROM A MARINE CYANOBACTERIA 155 NEW AROMATIC BISABOLANE DERIVATIVES WITH LIPID REDUCING ACTIVITY FROM THE MARINE SPONGE MYRMEKIODERMA SP 170 COMMERCIAL FUCOIDANS FROM FUCUS VESICULOSUS CAN BE GROUPED INTO ANTIADIPOGENIC AND ADIPOGENIC AGENTS ABSTRACT FUCUS VESICULOSUS IS A BROWN SEAWEED USED IN THE TREATMENT OF OBESITY THIS SEAWEED SYNTHESIZES VARIOUS BIOACTIVE MOLECULES ONE OF THEM BEING A SULFATED POLYSACCHARIDE KNOWN AS FUCOIDAN FF THIS POLYMER CAN EASILY BE FOUND COMMERCIALLY AND HAS ANTIADIPOGENIC AND LIPOLYTIC ACTIVITY USING DIFFERENTIAL PRECIPITATION WITH ACETONE WE OBTAINED FOUR FUCOIDAN RICH FRACTIONS F0 5 F0 9 F1 1 F2 0 FROM FF THESE FRACTIONS CONTAIN DIFFERENT PROPORTIONS OF FUCOSE GLUCURONIC ACID GALACTOSE XYLOSE SULFATE AND ALSO SHOWED DIFFERENT ELECTROPHORETIC MOBILITY AND ANTIOXIDANT ACTIVITY USING 3T3 L1 ADIPOCYTES WE FOUND THAT ALL SAMPLES HAD LIPOLYTIC ACTION ESPECIALLY F2 0 WHICH TRIPLED THE AMOUNT OF GLYCEROL IN THE CELLULAR MEDIUM MOREOVER WE OBSERVED THAT FF F1 0 AND F2 0 HAVE ANTIADIPOGENIC ACTIVITY AS THEY INHIBITED THE OIL RED STAINING BY CELLS AT 40 40 AND 50 RESPECTIVELY IN ADDITION THEY DECREASED THE EXPRESSION OF KEY PROTEINS OF ADIPOGENIC DIFFERENTIATION C EBP C EBP AND PPAR HOWEVER F0 5 AND F0 9 STIMULATED THE OIL RED STAINING AT 80 AND INCREASED THE EXPRESSION OF THESE PROTEINS THEREFORE THESE FUCOIDAN FRACTIONS HAVE AN ADIPOGENIC EFFECT OVERALL THE DATA SHOW THAT F2 0 HAS GREAT POTENTIAL TO BE USED AS AN AGENT AGAINST OBESITY AS IT DISPLAYS BETTER ANTIOXIDANT LIPOLYTIC AND ANTIADIPOGENIC ACTIVITIES THAN THE OTHER FUCOIDAN FRACTIONS THAT WE TESTED KEYWORDS 3T3 L1 CELLS FUCAN LIPOLYTIC OBESITY BROWN SEAWEED 1 INTRODUCTION FUCUS VESICULOSUS IS A BROWN SEAWEED COMMONLY FOUND IN COASTAL WETLANDS IN TEMPERATE OR COLD WATERS OF THE ATLANTIC AND PACI C OCEANS IT WAS SHOWN THAT F VESICULOSUS INTAKE HELPS WOMEN WITH ABNORMAL MENSTRUAL CYCLES AND HEALTH PROBLEMS ASSOCIATED WITH THEIR PERIODS 1 OTHER AUTHOR ALSO REPORTED THAT CONSUMPTION OF THIS SEAWEED PROMOTES A DECREASE IN BODY WEIGHT 2 3 THIS SEAWEED HAS VARIOUS ACTIVE ELEMENTS IN ITS COMPOSITION OF WHICH FUCOIDAN IS ONE OF THE BEST KNOWN THE PRESENCE OF FUCOIDAN IN F VESICULOSUS WAS DEMONSTRATED IN 1913 AND WAS INITIALLY CALLED FUCOIDIN 4 YEARS LATER IT WAS SUGGESTED THAT THE TERM BE CHANGED TO FUCOIDAN 5 1 THE STRUCTURE OF FUCOIDAN FF FROM F VESICULOSUS WAS LAST REVIEWED BY PATANKAR ET AL 6 IT WAS SUGGESTED THAT IT POSSESSES A CENTRAL CORE FORMED BY L FUCOSE 1 3 LINKED SULFATED AT C4 IN ADDITION SEVERAL BRANCHING POINTS EVERY TWO OR THREE FUCOSE RESIDUES WERE PRESENT IN 1 2 OR 1 4 LINKED ON THE MAIN CHAIN CURRENTLY IT IS EASY TO ACQUIRE FUCOIDAN FROM F VESICULOSUS AS A MULTINATIONAL COMPANY SELLS IT COMMERCIALLY IN PART THIS MAY EXPLAIN THE LARGE AMOUNT OF RESEARCH AND THE NUMBER OF ACTIVITIES ASCRIBED TO THIS SULFATED POLYSACCHARIDE INCLUDING ANTILIPEMIC 7 8 AND ANTIADIPOGENIC ACTIVITIES 9 10 ADIPOGENESIS IS A PROCESS OF CELL DIFFERENTIATION IN WHICH MESENCHYMAL STEM CELLS DIFFERENTIATE INTO ADIPOCYTES WITH THIS PROCESS INITIALLY INVOLVING A STAGE WHERE CELLS COMPROMISE WITH THE ADIPOCYTE LINE 11 IN THE NEXT STEP DIFFERENTIATION OCCURS AND THE PRE ADIPOCYTES DEVELOP INTO MATURE ADIPOCYTES THE PROCESS OF DIFFERENTIATION IS COMPLEX AND INVOLVES THE PARTICIPATION OF HUNDREDS OF PROTEINS ALTHOUGH TWO PROTEINS PARTICULARLY PLAY A CRUCIAL ROLE IN THIS EVENT C EBP A PROTEIN OF THE CCAAT ENHANCER BINDING PROTEIN CLASS AND PPAR A PEROXISOME PROLIFERATOR ACTIVATED RECEPTOR 12 ALTHOUGH PRE ADIPOCYTE PRIMARY CULTURES ARE AN IMPORTANT TOOL FOR UNDERSTANDING THE MECHANISMS OF ADIPOCYTE DIFFERENTIATION THESE CELLS HAVE LOW MITOGENIC CAPACITY AND LOSE THEIR ABILITY TO DIFFERENTIATE OVER TIME UNDER CULTURE CONDITIONS 13 THEREFORE THE DEVELOPMENT OF STUDIES ON ADIPOGENESIS IS CARRIED OUT MAINLY BY THE USE OF CELLULAR MODELS SUCH AS THE MURINE 3T3 L1 AND 3T3 F442A PRE ADIPOCYTE LINES THESE CELLS WHEN STIMULATED TO DIFFERENTIATE INTO ADIPOCYTES FOLLOW THE SAME METABOLIC PATHWAYS OF DIFFERENTIATION OF MESENCHYMAL CELLS 14 BY USING A DIFFERENTIATION COCKTAIL BASED ON INSULIN DEXAMETHASONE ISOBUTYLMETHYLXANTHINE AND FETAL BOVINE SERUM IT IS POSSIBLE TO OBTAIN MATURE ADIPOCYTES FROM A 3T3 L1 CULTURE THE ACTIONS OF THESE COMPOUNDS RESULT IN THE INITIAL EVENTS OF DIFFERENTIATION REPRESENTED BY THE EXPRESSION OF CCAAT ENHANCER BINDING PROTEINS 13 15 AFTERWARDS THE CELLS RETURN TO THEIR CELL CYCLES UNDERGO CLONAL EXPANSION IN A REGULATED MANNER AND ENTER A TERMINAL DIFFERENTIATION PROCESS BY ACTIVATION OF PPAR AND C EBP 16 BESIDES THE DIFFERENTIATION OF THESE CELLS TAKES PLACE IN PRECISELY CONTROLLED SEQUENTIAL STAGES CELL CYCLE ARREST CLONAL EXPANSION AND DIFFERENTIATION RST PHASE AND SECOND PHASE OF ACTIVATION OF TRANSCRIPTIONAL FACTORS BY ACTIVATING HUNDREDS OF PREVIOUSLY SILENCED GENES 15 STUDIES WITH PRE ADIPOCYTE 3T3 L1 CELLS SHOWED THAT FUCOIDAN FROM F VESICULOSUS INHIBITS ADIPOGENESIS REAL TIME POLYMERASE CHAIN REACTION PCR DATA SHOWED THAT FUCOIDAN REDUCED MRNA EXPRESSION OF C EBP AND PPAR BY 22 6 AND 17 6 RESPECTIVELY 9 FF FROM F VESICULOSUS CAN BE FRACTIONATED WITH CERTAIN FRACTIONS SHOWING VERY SIMILAR ACTIVITIES TO EACH OTHER 10 MOREOVER NISHINO ET AL 17 REPORTED THAT SOME FUCOIDAN FRACTIONS SHOWED MUCH GREATER ACTIVITY THAN OTHERS DID HOWEVER ANTIADIPOGENIC ACTIVITY ACROSS DIFFERENT FUCOIDAN POPULATIONS HAS NOT YET BEEN EVALUATED WITH THIS IN MIND WE OBTAINED FOUR DIFFERENT FUCOIDAN RICH FRACTIONS OF COMMERCIAL FUCOIDAN FROM F VESICULOSUS AND ASSESSED THEM FOR THEIR ADIPOGENIC ACTIVITY 2 RESULTS 2 1 OBTAINING DIFFERENT FRACTIONS OF FUCOIDAN FF USING DIFFERENTIAL PRECIPITATION WITH ACETONE WE OBTAINED FOUR FRACTIONS FROM FF THESE WERE CALLED F0 5 F0 9 F1 1 AND F2 0 CORRESPONDING TO 4 5 35 2 22 0 AND 38 3 OF THE MATERIAL RESPECTIVELY TABLE 1 CHEMICAL ANALYSIS AND SULFATED POLYSACCHARIDE YIELD ARE SUMMARIZED IN TABLE 1 DATA SHOW THAT MANNOSE AND GLUCOSE WERE NOT FOUND IN THE SAMPLES WHEREAS FUCOSE GLUCURONIC ACID GALACTOSE AND XYLOSE WERE FOUND IN ALL SAMPLES THE DATA ALSO SHOWED FUCOSE WAS THE MAJOR COMPONENT PRESENT IN ALL FRACTIONS WHEREAS THE RELATIVE AMOUNTS OF OTHER MONOSACCHARIDES VARY ACCORDING TO THE FRACTION THUS THE RELATIVE AMOUNTS OF THESE SUGARS VARY ACCORDING TO THE FRACTION WHEN THE SULFATE CONTENT OF THE SAMPLES WAS QUANTI ED IT WAS OBSERVED THAT THERE IS NO SIGNI CANT DIFFERENCE BETWEEN F1 1 AND F2 0 ALTHOUGH F1 1 AND F2 0 BOTH HAVE THE SAME SULFATE CONTENT THEY WERE PRECIPITATED WITH DIFFERENT VOLUMES OF ACETONE THIS IS PROBABLY BECAUSE THE SULFATED POLYSACCHARIDE CONFORMATION INTERFERES DURING THE PRECIPITATION PROCESS 2 TO CONFIRM THIS HYPOTHESIS THE FUCOIDAN RICH FRACTIONS WERE SUBJECTED TO AGAROSE GEL ELECTROPHORESIS IN 1 3 DIAMINEACETATE PDA BUFFER FIGURE 1 SHOWS AN AGAROSE GEL STAINED WITH TOLUIDINE BLUE IT IS POSSIBLE TO SEE THAT ALL THE FRACTIONS HAVE A PREDOMINANT BAND FOR F0 9 F1 1 AND F2 0 THE BANDS DISPLAY DIFFERENT ELECTROPHORETIC MOBILITIES TABLE 1 CHEMICAL COMPOSITION OF FUCOIDAN FF AND ITS FRACTIONS FUC FUCOSE GLUC ACID GLUCURONIC ACID GAL GALACTOSE XYL XYLOSE MAN MANNOSE GLUC GLUCOSE N D NOT DETECTED DIFFERENT LETTERS A B C D INDICATE A SIGNI CANT DIFFERENCE P 0 05 BETWEEN THE SAMPLES EACH VALUE IS THE MEAN STANDARD DEVIATION SD OF THREE DETERMINATIONS AND FROM THREE INDEPENDENT ASSAYS SULFATED POLYSACCHARIDES YIELD SULFATE MOLAR RATIO FUC GLUC ACID GAL XYL MAN GLUC FF 23 70 0 04 A 1 0 0 7 0 4 0 2 N D N D F0 5 4 5 12 70 0 08 B 1 0 0 2 0 3 0 5 N D N D F0 9 35 2 17 40 0 02 C 1 0 0 8 0 2 0 6 N D N D F1 1 22 0 20 30 0 05 D 1 0 0 2 0 4 0 2 N D N D F2 0 38 3 20 40 0 04 D 1 0 0 1 0 3 0 1 N D N D FIGURE 1 STAINING PATTERN OF THE POLYSACCHARIDES AFTER AGAROSE GEL ELECTROPHORESIS STAINED WITH TOLUIDINE BLUE ABOUT 5 L 50 G OF EACH SAMPLE WAS APPLIED IN AGAROSE GEL PREPARED IN DIAMINOPROPANE ACETATE BUFFER AND SUBJECTED TO ELECTROPHORESIS AS DESCRIBED IN METHODS OR ORIGIN THIS FIGURE IS REPRESENTATIVE OF THREE SEPARATE TESTS MADE INDEPENDENTLY 2 2 ANTIOXIDANT ACTIVITIES THE ANTIOXIDANT ACTIVITY OF SAMPLES WAS EVALUATED IN VITRO BY THE TOTAL ANTIOXIDANT CAPACITY TEST TAC ALL SAMPLES SHOWED ANTIOXIDANT ACTIVITY THOUGH THE VALUES WERE SIGNIFICANTLY DIFFERENT FROM EACH OTHER PARTICULARLY FOR F2 0 FUCOIDAN WHICH WAS APPROXIMATELY 400 12 0 EQUIVALENTS OF ASCORBIC ACID THIS IS NEARLY DOUBLE THE VALUES OBTAINED FOR FF AND F0 9 WHICH WERE 189 10 0 AND 172 11 0 ASCORBIC ACID EQUIVALENTS RESPECTIVELY THE VALUE IDENTIFIED FOR F1 1 WAS FOUND TO BE SIGNIFICANTLY LOWER THAN THE TWO MENTIONED ABOVE 150 8 0 EQUIVALENTS THE VALUE OBTAINED FOR F0 5 WAS AROUND 38 2 0 EQUIVALENTS THE LOWEST RECORDED IN THIS STUDY THERE WAS POSITIVE PEARSON CORRELATION COEF CIENT BETWEEN THE SULFATED CONTENT OF FRACTIONS AND TAC P 0 566 3