Search

Intergenerational climate justice negotiations often flounder on three questions: What was the outcome on climate change? Was it intentional? Why should the current generation pay for the misdeeds of previous generations? In this research, participants from Japan and Canada rated their willingness to accept intergenerational climate legacies and the responsibilities these legacies entail; judged the importance of intent and outcome associated with creating these legacies; and rated their willingness to compensate those negatively impacted by previous generations. The study found: a) while outcome was important, intent did not matter; b) Canadians were more likely to accept an inheritance and c) more likely to equivocate, in acceptance, if it entailed obligations than the Japanese; d) among those who accepted the inheritance, Japanese were more generous in settlement of previous generation’s obligations; e) lower-income, non-Judeo-Christian participants were systematically fairer than others; and f) the resistance to compensation for past generations’ actions was diminished with the awareness about the broad scope of intergenerational climate legacies that the current generation enjoyed. Our findings highlight the influences of culture and historic awareness on accepting climate responsibilities for actions of previous generations and willingness to provide compensation. The findings also support abandoning the debate on intentionality.

FMS-like tyrosine kinase 3 (FLT3) is one of the most frequently mutated genes in acute myeloid leukemia (AML), and its activating mutations including internal tandem duplications (ITDs) and missense point mutations in the tyrosine kinase domain (TKD) are found in approximately 30% of AML patients. Although the patients initially well responded to FLT3 inhibitors such as gilteritinib, the most cases relapsed within a few months after the initiation of treatment. For these patients who relapsed after FLT3 inhibitor-based therapy, no effective treatment is available. Mechanisms of resistance are not fully elucidated, but FLT3-ITD-TKD double mutant is considered as one of possible mechanisms. We have discovered a novel FLT3 inhibitor OTS447 that shows the potent and selective inhibition in both FLT3-ITD and FLT3-ITD-TKD double mutants. OTS447 was identified through the screening of our in-house proprietary compound library. It showed potent inhibitory activity against FLT3 with an IC50 value of 0.19 nM. The selectivity of OTS447 was investigated by human kinase profile assay. Among 371 human kinases tested, there were only seven (including FLT3) whose activity were inhibited by 80% or more at 5 nM of OTS447. We then examined the anti-proliferative effects in FLT3-ITD and -wild type (WT) AML cell lines. The proliferation of FLT3-ITD cell lines, MV4-11 and MOLM13, was more strongly suppressed than that of FLT3-WT cell lines. We also investigated the inhibitory effects to FLT3-ITD-TKD double mutants using Ba/F3 cells. FLT3-ITD-D835Y mutant was inhibited as strongly as FLT3-ITD mutant, and FLT3-ITD-F691I mutant was more strongly inhibited than Ba/F3 parental cells. These data suggest that OTS447 has selective inhibitory activity against FLT3 mutants. To confirm that FLT3 inhibition by OTS447 leads to the anti-proliferative effect, FLT3 autophosphorylation and phosphorylation of downstream molecules such as STAT5, ERK and AKT were examined. Both autophosphorylation of FLT3 and phosphorylation of downstream molecules were decreased by OTS447 in dose-dependent manner. Moreover, treatment of OTS447 induced the apoptosis and increased sub-G1 population in MV4-11 cell. Finally, we tested OTS447 anti-tumor effect using MV4-11 mouse xenograft model. OTS447 showed potent growth inhibition against MV4-11 tumor in dose-dependent manner. In summary, we discovered OTS447, a potent and selective FLT3 inhibitor, that can inhibit not only ITD mutation but also ITD-TKD mutations of the FLT3 gene. OTS447 possesses cytotoxic activity induced by inhibition of FLT3 signaling pathway and has anti-tumor activity in mouse xenograft model. We are pursuing further optimization of the inhibitor aiming for the effective treatment of AML patients with several types of FLT3-activating mutations. Citation Format: Yasuhide Okamoto, Naofumi Takamatsu, Takashi Miyamoto, Akiko Taira, Kozue Toyota, Yukiho Imai, Shinobu Fujii, Rumiko Ono, Kyoko Adachi, Yo Matsuo, Suyoun Chung, Jae-Hyun Park. Discovery of OTS447, a highly potent and selective inhibitor of FLT3 for the treatment of AML patients with FLT3-ITD/TKD mutations [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5456.

A Gram-stain-negative, strictly aerobic, pale-yellow pigmented, rod-shaped, chemoheterotrophic bacterium, designated A6F-11(T), was isolated from a marine sponge collected in Japan. Phylogenetic analysis based on the 16S rRNA gene sequence indicated that the novel marine strain was affiliated with the family Flavobacteriaceae of the phylum Bacteroidetes and that it shared the highest (92.9 %) sequence similarity with Arenibacter palladensis LMG 21972(T). The strain could be differentiated phenotypically from related members of the family Flavobacteriaceae. The major fatty acids of strain A6F-11(T) were iso-C15:1 G, iso-C15:0, C16:1 ω6c and/or C16:1 ω7c and iso-C17:0 3-OH. The polar lipid profile consisted of phosphatidylglycerol, two unidentified aminolipids and two unidentified lipids. The DNA G+C content was 34.7 mol%, and the major respiratory quinone was menaquinone 6 (MK-6). From the distinct phylogenetic position and combination of genotypic and phenotypic characteristics, the strain is considered to represent a novel taxon in the family Flavobacteriaceae, for which the name Spongiimicrobium salis gen. nov., sp. nov. is proposed. The type strain of S. salis gen. nov., sp. nov. is A6F-11(T) (= KCTC 42753(T) = NBRC 111401(T)).

Random mutagenesis for the hyperthermophilic archaeon Thermococcus kodakarensis was established by the insertion of an artificial transposon designed to allow easy identification of the transposon-inserted locus. The phenotypic screening was applied for the isolation of thermosensitive mutants of T. kodakarensis, which resulted in the isolation of 16 mutants showing defective growth at the supraoptimal temperature 93°C. The high occurrence of the mutants suggested that the high thermotolerance of hyperthermophiles was achieved by a combination of diverse gene functions. The transposon insertion sites in two-thirds of the mutants were identified in a group of genes responsible for tRNA modifications including 7-formamidino-7-deaza-guanosine (archaeosine), N1-methyladenosine/N1-methylinosine, N4-acetylcytidine, and N2-dimethylguanosine/N2,N2-dimethylguanosine. LC–MS/MS analyses of tRNA nucleosides and fragments exhibited disappearance of the corresponding modifications in the mutants. The melting temperature of total tRNA fraction isolated from the mutants lacking archaeosine or N1-methyladenosine/N1-methylinosine decreased significantly, suggesting that the thermosensitive phenotype of these mutants was attributed to low stability of the hypomodified tRNAs. Genes for metabolism, transporters, and hypothetical proteins were also identified in the thermosensitive mutants. The present results demonstrated the usefulness of random mutagenesis for the studies on the hyperthermophile, as well as crucial roles of tRNA modifications in cellular thermotolerance.

Jasmonates regulate plant defense and development. In Arabidopsis (Arabidopsis thaliana), JASMONATE-ASSOCIATED VQ-MOTIF GENE1 (JAV1/VQ22) is a repressor of jasmonate-mediated defense responses and is degraded through the ubiquitin-26S proteasome system after herbivory. We found that JAV1-ASSOCIATED UBIQUITIN LIGASE1 (JUL1), a RING-type E3 ubiquitin ligase, interacted with JAV1. JUL1 interacted with JAV1 in the nucleus to ubiquitinate JAV1, leading to proteasomal degradation of JAV1. The transcript levels of JUL1 and JAV1 were coordinately and positively regulated by the CORONATINE INSENSITIVE1-dependent signaling pathway in the jasmonate signaling network, but in a manner that was not dependent on CORONATINE INSENSITIVE1-mediated signaling upon herbivory by Spodoptera litura. Gain or loss of function of JUL1 modulated the expression levels of the defensin gene PDF1.2 in leaves, conferring on the plants various defense properties against the generalist herbivore S. litura. Because neither the JUL1 mutant nor overexpression lines showed any obvious developmental defects, we concluded that the JAV1/JUL1 system functions as a specific coordinator of reprogramming of plant defense responses. Altogether, our findings offer insight into the mechanisms by which the JAV1/JUL1 system acts specifically to coordinate plant defense responses without interfering with plant development or growth.

Phylogenetic and taxonomic characterization was performed for a bacterium, designated strain Q-192T, isolated from the surface of the green macroalga Halimeda sp., collected from the subtropical Ishigaki Island, Japan. The isolate was a polysaccharide-producing, Gram-stain-negative, aerobic, rod-shaped, motile bacterium with a polar flagellum. The isolate was slightly halophilic, required Na+, Mg2+ and Ca2+ ions for growth, but did not require growth factors. The only isoprenoid quinone was ubiquinone-8.The major cellular fatty acids were C18 : 1ω7c, C16 : 0 and C14 : 0. The main hydroxy fatty acid was C10 : 0 3-OH. The DNA G+C content was 45.9 mol%. Phylogenetic analysis of 16S rRNA gene sequences placed the isolate in the class Gammaproteobacteria. The phylogenetically closest relatives with validly published names were Pseudomaricurvus alkylphenolicus KU41GT, Teredinibacter turnerae T7902T, Pseudoteredinibacter isoporae SW-11T and Simiduia agarivorans SA1T with sequence similarities of 94.5, 94.1, 93.7 and 93.6 %, respectively. The isolate was distinguished from members of these genera by a combination of DNA G+C content, chemotaxonomic characteristics (respiratory quinone system, fatty acid profile and polar lipid composition) and other phenotypic features. Based on phylogenetic, genotypic, chemotaxonomic and phenotypic characteristics, strain Q-192T is considered to represent a novel species of a new genus, for which the name Marinibactrum halimedae gen. nov., sp. nov. is proposed. The type strain of Marinibactrum halimedae is Q-192T ( = NBRC 110095T = NCIMB 14932T).

A facultatively anaerobic, Gram-stain negative, golden-yellow pigmented, non-motile and rod-shaped bacterium, designated strain A5Q-67(T) was isolated from leaf litter collected at the mangrove estuary of Nakama River, Japan. Phylogenetic analysis based on the 16S rRNA gene sequence revealed the novel isolate was affiliated with the family Flavobacteriaceae of the phylum Bacteroidetes and that it showed highest sequence similarity (94.2 %) to Imtechella halotolerans K1(T). The strain could be differentiated phenotypically from recognized members of the family Flavobacteriaceae. The major fatty acids of strain A5Q-67(T) were identified as iso-C17:0 3-OH, summed feature 1 (iso-C15:1 H and/or C13:0 3-OH) and iso-C15:0 as defined by the MIDI system. The DNA G+C content was determined to be 36.7 mol%, the major respiratory quinone was identified as menaquinone 6 (MK-6) and a polar lipid profile was present consisting of phosphatidylethanolamine, two unidentified aminolipids and an unidentified lipid. From the distinct phylogenetic position and combination of genotypic and phenotypic characteristics, the strain is considered to represent a novel genus for which the name Frondibacter aureus gen. nov., sp. nov. is proposed. The type strain of F. aureus is A5Q-67(T) (=KCTC 32991(T) = NBRC 110021(T)).

A Gram-stain-negative, rod-shaped, non-motile, strictly aerobic, chemoheterotrophic, pale-yellow-pigmented bacterium, designated strain A6D-50T, was isolated from an ascidian collected at Kohama Island offshore Okinawa, Japan. Preliminary analysis based on the 16S rRNA gene sequence revealed that the novel isolate was affiliated with the family Flavobacteriaceae of the phylum Bacteroidetes and that it showed highest sequence similarity (97.6 %) to Aureisphaera galaxeae 04OKA003-7T. The DNA-DNA relatedness value between strain A6D-50T and A. galaxeae 04OKA003-7T was 23.6 %. The DNA G+C content of strain A6D-50T was 40.8 mol%, MK-6 was the only menaquinone, and iso-C17 : 0 3-OH, iso-C15 : 0, iso-C15 : 1 G and summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c) were the major (>10 %) cellular fatty acids. The polar lipid profile consisted of phosphatidylethanolamine, two unidentified aminolipids and an unidentified lipid. From the distinct phylogenetic position and combination of genotypic and phenotypic characteristics, the strain is considered to represent a novel species of the genus Aureisphaera for which the name Aureisphaera salina sp. nov. is proposed. The type strain is A6D-50T (= KCTC 42975T=NBRC 111827T).

A new rifamycin antibiotic, salinisporamycin (1), has been isolated from a culture of a marine actinomycete. The producing organism was identified as Salinispora arenicola [corrected] on the basis of the 16S rRNA sequence. High-resolution FAB-MS established the molecular formula of 1 as C(33)H(43)NO(9). The planar structure of 1 was elucidated by NMR spectral analysis including COSY, heteronuclear single quantum coherence and heteronuclear multiple bond correlation. The relative stereochemistry of 1 was determined on the basis of rotating frame nuclear Overhauser effect spectroscopy. In addition, the solvatochromic behavior of 1 was investigated by measuring the UV spectra. This compound inhibited the growth of A549 cells, the human lung adenocarcinoma cell line, with an IC(50) value of 3 microg ml(-1), and also showed antimicrobial activity.

Two marine heterotrophic bacteria, A5K-61(T) and A5K-106(T), were isolated from marine animals. 16S rRNA gene sequence analysis data showed that the isolates were affiliated with the genus Thalassomonas; highest 16S rRNA gene sequence similarity values were found with Thalassomonas viridans DSM 13754(T) (97.5 and 98.1 %, respectively). DNA-DNA hybridization values of strains A5K-61(T) and A5K-106(T) with T. viridans DSM 13754(T) (22.2-49.1 %) were clearly below 70 %, the generally accepted limit for species delineation. The isolates produced a brown diffusible pigment. The major respiratory quinone was Q-8 and the predominant cellular fatty acids were C(16 : 1)omega7c and C(16 : 0). Based on DNA-DNA hybridization data, some biochemical characteristics and 16S rRNA gene sequence comparison, the isolates represent two novel species of the genus Thalassomonas, for which the names Thalassomonas actiniarum sp. nov. (type strain A5K-106(T) =MBIC08328(T) =NCIMB 14418(T) =NBRC 104231(T)) and Thalassomonas haliotis sp. nov. (type strain A5K-61(T) =MBIC08329(T) =NCIMB 14417(T) =NBRC 104232(T)) are proposed.

Suppressive effects of neocarrabiose 4-O-sulfate on the thermal and freeze denaturation of myosin subfragment-1 (S-1) and on myofibrils were investigated. The compound strongly suppressed the thermal denaturation of S-1. Its suppressive effect was greater than that of sorbitol and similar to that of maltose. However, it tended to accelerate the denaturation of myofibrils, suggesting a loss of protection by F-actin upon its addition. The compound suppressed the freeze denaturation of myofibrils and S-1. The effect was similar to that of sorbitol or maltose, and completely different from that of Na-sulfate. The compound solubilized myofibrils at concentrations similar to KCl. Therefore, it was concluded that neocarrabiose 4-O-sulfate behaved as an ionic salt in the thermal treatment process, whereas it behaved as a sugar in the frozen storage process.

Two strains of marine, heterotrophic, alkaliphilic bacteria, designated A3F-7(T) and ssthio04PA2-7c, were isolated from hard coral and marine sand, respectively, collected in the Republic of Palau. A phylogenetic analysis based on 16S rRNA gene sequences showed that the isolates were related to members of the genus Moritella. However, the sequence similarities between the isolates and the type strains of the Moritella species were less than 93 %. The G+C contents of the isolates were around 57 mol%, the major respiratory quinone was Q-8 and the predominant cellular fatty acids were 16 : 1omega7c, 16 : 0, 18 : 1omega7c and 14 : 0. On the basis of the phylogenetic data, phenotypic characteristics and DNA-DNA hybridization results, strains A3F-7(T) and ssthio04PA2-7c represent a novel species of a novel genus, for which the name Paramoritella alkaliphila gen. nov., sp. nov. is proposed. The type strain of Paramoritella alkaliphila is A3F-7(T) (=MBIC06429(T) =DSM 19956(T)).

Chalcone compounds have been widely studied for their anti-inflammatory, anti-pyretic, anti-invasive and anti-proliferative activities in various cell lines. However, their effects on the central nervous system (CNS) are still largely unexplored. We have recently developed a bioconversion system using a recombinant Escherichia coli that enables us to produce chemical compounds that are naturally rare and usually difficult to chemically synthesize. One such compound is 3-(2,3-dihydroxyphenyl)-1-phenylpropan-1-one, a novel chalcone-diol. Here we show, for the first time, that the chalcone-diol enhanced the phosphorylation of extracellular signal-regulated kinase (ERK) in a time- and concentration-dependent manner in cultured cortical neurons. Also, this chalcone-diol increased intracellular cyclic AMP (cAMP) concentration, thereby enhancing phosphorylation of ERK and cAMP-response element-binding protein (CREB), and CRE-mediated transcription via the cAMP-dependent protein kinase (PKA)/mitogen-activated protein kinase/ERK kinase (MEK) pathway in cultured rat hippocampal neurons. Recent studies have demonstrated that PKA/CREB-dependent signaling, which is required for long-term potentiation, is inhibited by sublethal concentrations of amyloid beta-peptide (Abeta) in cultured hippocampal neurons. After treatment with the chalcone-diol at 50 muM prior to treatment with a sublethal concentration of Abeta(1-42), the Abeta(1-42)-induced inhibition of phosphorylation of PKA substrates and CREB was prevented in cultured hippocampal neurons, indicating the potential for protection against the Abeta-induced impairment of PKA/CREB signaling observed in Alzheimer's disease. Therefore, these results suggest that our present study provides a new approach for discovering novel lead compounds for the treatment of neurodegenerative CNS diseases associated with impaired PKA/CREB signaling, including Alzheimer's disease.

Four bacterial strains, 04HE-4-40, A4I-9, A4I-21 and AK15-027(T), were isolated from Heita Bay off Kamaishi in Japan. Based on 16S rRNA gene sequence analysis, the isolates were placed as a new lineage within the genus Psychromonas. The 16S rRNA gene sequence similarity values between the isolates and type strains of recognized Psychromonas species were less than 97 %. The DNA G+C contents of the isolates were about 38 mol%. The major respiratory quinone was Q-8, and the predominant cellular fatty acids were 16 : 1omega7c and 16 : 0. The isolates were able to grow at 30 degrees C, unlike other Psychromonas species. Based on the results of the phylogenetic analysis and phenotypic characteristics and DNA-DNA hybridization data, the isolates represent a novel species, for which the name Psychromonas heitensis sp. nov. is proposed. The type strain is AK15-027(T) (=MBIC 06898(T)=NCIMB 14416(T)).

Ariakemicins A (1) and B (2), unusual linear hybrid polyketide-nonribosomal peptide antibiotics, were discovered from the fermentation extract of the marine gliding bacterium Rapidithrix sp. These metabolites were positional isomers with regard to a double bond and chromatographically inseparable, rendering the structure study on a mixture basis. The ariakemicins were composed of threonine, two omega-amino-(omega-3)-methyl carboxylic acids with diene or triene units, and delta-isovanilloylbutyric acid. The antibiotics selectively inhibited the growth of Gram-positive bacteria.

Ten pale-pink- and pale-yellow-pigmented, Gram-negative, non-motile, rod-shaped, chemoheterotrophic bacteria designated strains YM20-087T, YM21-151, MN1-741T, YM27-120T, YM26-010T, YM24-184, YM20-122, A4T-83T, A5J-41-2T and A5J-40 were isolated from various marine environments and were subjected to a polyphasic taxonomic investigation. Phylogenetic analyses based on 16S rRNA gene sequences indicated that these isolates belonged to the phylum 'Verrucomicrobia' (subdivision 1) and represented three independent lineages that were distinct from species of genera of the family Verrucomicrobiaceae with validly published names. The cell-wall peptidoglycan of these strains contained muramic acid and meso-diaminopimelic acid. Strains MN1-741T, YM27-120T, YM26-010T, YM24-184 and YM20-122 produced pinkish carotenoid pigments. On the basis of polyphasic taxonomic evidence, it was concluded that these strains should be classified within three new genera, Persicirhabdus gen. nov. (with one species, the type species Persicirhabdus sediminis sp. nov.), Roseibacillus gen. nov. (with three species; type species Roseibacillus ishigakijimensis sp. nov.) and Luteolibacter gen. nov. (with two species; type species Luteolibacter pohnpeiensis sp. nov.), of the family Verrucomicrobiaceae within the phylum 'Verrucomicrobia'. The names Persicirhabdus sediminis gen. nov., sp. nov. (type strain YM20-087T =MBIC08313T =KCTC 22039T), Roseibacillus ishigakijimensis gen. nov., sp. nov. (type strain MN1-741T =MBIC08315T =KCTC 12986T), Roseibacillus ponti sp. nov. (type strain YM27-120T =MBIC08316T =KCTC 12987T), Roseibacillus persicicus sp. nov. (type strain YM26-010T =MBIC08317T =KCTC 12988T), Luteolibacter pohnpeiensis gen. nov., sp. nov. (type strain A4T-83T =MBIC08322T =KCTC 22041T) and Luteolibacter algae sp. nov. (type strain A5J-41-2T =MBIC08320T =KCTC 22040T) are therefore proposed. Emended descriptions of the class Verrucomicrobiae, the order Verrucomicrobiales and the family Verrucomicrobiaceae are also presented.

Shampoo ginger (Zingiber zerumbet Smith) has a high content and large variety of terpenoids in the essential oil of its rhizome. Here, we report on the isolation of a cDNA clone (ZSS1) encoding alpha-humulene synthase, a possible key enzyme of zerumbone biosynthesis. This clone contains an open reading frame of 1,644 bp and is predicted to encode a protein of 548 amino acids with a calculated molecular mass of 64.5 kDa. The deduced amino acid sequence shows 34-63% identity with known sesquiterpene synthases of other angiosperm species. Based on exon-intron organization, ZSS1 is classified as the terpene synthase-III (TPS-III) subfamily. When expressed in Escherichia coli, the recombinant enzyme catalyzed the formation of a major product, alpha-humulene (95%) and a minor by-product, beta-caryophyllene (5%). Introduction of ZSS1 and the foreign mevalonate pathway involved in FPP synthesis into E. coli results in in vivo production of alpha-humulene. Transcript of ZSS1 was detected almost exclusively in rhizomes and was up-regulated in both leaves and rhizomes after treatment with methyl jasmonate (MeJA), suggesting its ecological function in shampoo ginger.

Two new antibiotic depsipeptides, unnarmicins C (1) and A (2), were isolated from the fermentation broth of a marine bacterium, Photobacterium sp. strain MBIC06485. The structure of 1 was established by spectroscopic studies and chiral analyses of its chemical degradation/conversion products, and that of 2 by comparing its NMR, MS, and CD data with those of 1. Both compounds selectively inhibited the growth of two strains belonging to the genus Pseudovibrio, one of the most prevalent genera in the marine environments within the class Alphaproteobacteria.

Novel inhibitors of fungal ATP-binding cassette transporters were obtained by screening compounds and crude extracts from marine-derived fungi and bacteria using disk diffusion assays of Saccharomyces cerevisiae strains overexpressing a variety of fungal multi-drug efflux pumps. The cyclodepsipeptides unnarmicin A and unnarmicin C were able to sensitize cells overexpressing azole drug pumps ScPdr5p, CaCdr1p, CgCdr1p, and CgPdh1p to sub-MIC concentrations of fluconazole without affecting the growth of CaCdr2p and CaMdr1p overexpressing cells. Unnarmicin A and unnarmicin C were potent inhibitors of rhodamine 6G efflux of CaCdr1p expressing cells with IC50 values of 3.61 and 5.65 microM, respectively. They inhibited the in vitro CaCdr1p ATPase activity at IC50 values of 0.495 and 0.688 microM, respectively. And most importantly, they were able to sensitize azole-resistant Candida albicans clinical isolates to fluconazole. Unnarmicin A and unnarmicin C are candidate efflux pump inhibitors with the potential to be used as adjuvants for antifungal chemotherapy.

Two Gram-negative, non-motile, coccoid or rod-shaped, chemoheterotrophic bacteria designated strains YM21-132T and YM27-005T were isolated from marine animals, and were subjected to a polyphasic taxonomic examination. Phylogenetic analyses based on 16S rRNA gene sequences indicated that the two isolates belong to the genus Rubritalea of the phylum ‘Verrucomicrobia’ (subdivision 1). The novel isolates shared approximately 97–98 % sequence similarity with each other and showed 93–97 % similarity with Rubritalea species of the family Verrucomicrobiaceae. The level of DNA–DNA relatedness between strains YM21-132T and YM27-005T was less than 70 %, which is accepted as the phylogenetic definition of a species. Both strains produced reddish carotenoid pigments and squalene. The cell wall peptidoglycan of both strains contained muramic acid and meso-diaminopimelic acid. The G+C contents of the genomic DNA were 48.0 mol% (strain YM21-132T) and 50.3 mol% (strain YM27-005T). The presence of MK-8 and MK-9 as the major isoprenoid quinones, and iso-C14 : 0, iso-C16 : 0 and C16 : 1 ω7c as the major cellular fatty acids supported the identification of the two novel strains as members of the genus Rubritalea. On the basis of polyphasic taxonomic studies, it was concluded that these strains should be classified as representing two novel, separate species in the genus Rubritalea within the phylum ‘Verrucomicrobia’, for which the names Rubritalea spongiae sp. nov. (type strain YM21-132T=MBIC08281T=KCTC 12906T) and Rubritalea tangerina sp. nov. (type strain YM27-005T=MBIC08282T=KCTC 12907T) are proposed.

SUMMARY Using nuclear magnetic resonance spectroscopy, we identified and characterized accumulated compatible solutes in cells of the haptophyte alga Pavlova sp. strain CCMP504. The predominant organic solutes were d-1,4/2,5-cyclohexanetetrol (CHT), 1,3,5/2,4-cyclohexanepentol (CHP) and scyllo-inositol. We then profiled the intracellular organic solutes present in Pavlova sp. grown in medium with salinity ranging from 23 practical salinity units (PSU) (hyposaline) to 35 PSU (optimum salinity for growth), to 47 PSU (hypersaline). The results of these analyses reveal progressive accumulation of CHT and CHP in response to increasing growth medium salinity. We also observed altered accumulation of CHT and CHP in samples subjected to salinity shock. To further characterize the CHT and CHP biosynthesis in Pavlova sp., we carried out stable isotope feeding experiments. Specific labeling of CHT and CHP with d-13C-glucose suggested that d-glucose is a biosynthetic precursor of these cyclitols. The salinity-induced accumulation of CHT and CHP suggests that these cyclitols act as compatible solutes. Our results therefore provide new evidence supporting classification of CHP as a compatible solute.

Urukthapelstatin A, a novel cyclic peptide, was isolated from the cultured mycelia of marine-derived Thermoactinomycetaceae bacterium Mechercharimyces asporophorigenens YM11-542. The peptide was purified by solvent extraction, silica gel chromatography, ODS flash chromatography, and finally by preparative HPLC. Urukthapelstatin A dose-dependently inhibited the growth of human lung cancer A549 cells with an IC(50) value of 12 nM. Urukthapelstatin A also showed potent cytotoxic activity against a human cancer cell line panel.

Piericidins C7 and C8, two new members of the piericidin family, were isolated from a marine actinomycete. The producing organism was identified as Streptomyces sp. based on the cultural, biochemical and chemotaxonomical characteristics as well as the 16S rRNA sequence. Piericidins C7 and C8 showed selective cytotoxicity against rat glia cells transformed with the adenovirus E1A gene (IC50 C7: 1.5 nM, C8: 0.45 nM) and Neuro-2a mouse neuroblastoma cells (IC50 C7: 0.83 nM, C8: 0.21 nM).

A polyphasic taxonomic approach was applied to determine the taxonomic position of a hydrocarbon-degrading actinomycete, strain Hou_blueT, which was isolated from soil samples collected from an oil spring in Niigata, Japan. The results of 16S rRNA and gyrB gene sequence comparisons indicated that strain Hou_blueT represented a novel lineage in the suborder Corynebacterineae. Colonies were malachite green-like in colour on 1/10 trypticase soy agar and the cell morphology was coccoid in all growth phases. The cell-wall diamino acid and sugar indicated chemotype IV and variation A1γ. The sugars of the peptidoglycan were glycolated. The polar lipids were composed of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, phosphatidylinositol mannoside and some unspecified glycolipids. The organism contained two novel cyclic forms of menaquinone, smaragdiquinone A-8(H4, ω-cycl) and smaragdiquinone B-8(H4, dicycl). The major fatty acids were cis-9-18 : 1 (34.46 %) and 16 : 0 (25.1 %). Small amounts of 10-methyl-branched fatty acids were also present (10-methyl-17 : 0, 0.17 %), but not tuberculostearic acid (10-methyl-18 : 0), which has been shown to be present in all nocardiae. Gas-chromatographic analysis of the mycolic acid revealed a carbon-chain length of C43–C49. The DNA G+C content was 63.7 mol%. On the basis of phenotypic and phylogenetic distinctness, the organism is proposed to represent a novel genus and species, Smaragdicoccus niigatensis gen. nov., sp. nov., with the type strain Hou_blueT (=MBIC 06267T=DSM 44881T).

Strains YM18-098T and A5E-52T were both Gram-positive, aerobic, irregular rod-shaped bacteria, with lysine and ornithine as the diagnostic diamino acids of their peptidoglycans, respectively. The acyl type of the peptidoglycan in both cases was N-glycolyl. The major menaquinones were MK-11, -12 and -13. Mycolic acids were not detected. The G+C content of the DNA was 69–70 mol%. Comparative 16S rRNA studies revealed that the isolates belonged to the genus Microbacterium, that YM18-098T was closely related to the species Microbacterium lacticum and Microbacterium schleiferi, and that A5E-52T was closely related to the species Microbacterium aurum, Microbacterium aoyamense, Microbacterium deminutum and Microbacterium pumilum. DNA-DNA relatedness analysis showed that the isolated strains represented two separate genomic species. Based on both phenotypic and genotypic data, the following new species of the genus Microbacterium are proposed: Microbacterium flavum sp. nov. and Microbacterium lacus sp. nov., with the type strains YM18-098T (= MBIC08278T, DSM 18909T) and A5E-52T (= MBIC08279T, DSM 18910T), respectively.

A Gram stain-negative, strictly aerobic, brown-pigmented, non-motile, rod-shaped, chemoheterotrophic bacterial strain-designated A6F-119(T) was isolated from a marine sponge (Rhabdastrella sp.). Phylogenetic analyses based on the 16S rRNA gene sequence revealed that the new strain represented a member of the family Flavobacteriaceae of the phylum Bacteroidetes and that it showed highest sequence similarity (93 %) to Tenacibaculum maritimum NBRC 15946(T). The strain could be differentiated phenotypically from the recognized members of the family Flavobacteriaceae. The DNA G + C content of strain A6F-119(T) was determined to be 30.8 mol%; MK-6 was identified as the major menaquinone; and the presence of iso-C15:0, iso-C17:0 3-OH, and C16:1 ω7c and/or C16:1 ω6c as the major (>10 %) cellular fatty acids. A polar lipid profile was present consisting of phosphatidylethanolamine, an unidentified aminolipid, and three unidentified lipids. From the distinct phylogenetic position and combination of genotypic and phenotypic characteristics, the strain is considered to represent a novel genus for which the name Spongiiferula fulva gen. nov., sp. nov. is proposed. The type strain of S. fulva is A6F-119(T) (= KCTC 42752(T) = NBRC 111402(T)).

A polyphasic taxonomic investigation was performed on a bacterial strain, 38-Ka-2(T), which was isolated from the green alga Ulva pertusa Kjellman (Chlorophyta) in Hokkaido, Japan. The bacterial cells were observed to be golden-yellow coloured, Gram-negative, strictly aerobic, non-spore-forming, non-motile and rod-shaped. Phylogenetic analyses based on the 16S rRNA gene sequence indicated that the new strain is a member of the family Flavobacteriaceae within the phylum Bacteroidetes and that it shows high sequence similarity (94.8 %) to Aquimarina addita JC2680(T). The genomic DNA G+C content of strain 38-Ka-2(T) was determined to be 36 mol%; MK-6 was identified as the major menaquinone; and the presence of iso-C15:0, C18:0 and iso-C17:0 3-OH as the major (10 %) cellular fatty acids. The polar lipid profile was found to consist of phosphatidylethanolamine, an unidentified glycolipid and two unidentified lipids. From the distinct phylogenetic position and combination of genotypic and phenotypic characteristics, strain 38-Ka-2(T) is considered to represent a novel genus for which the name Algitalea ulvae gen. nov., sp. nov. is proposed. The type strain of A. ulvae is 38-Ka-2(T) (=KCTC 32994(T) = NBRC 110017(T)).

Chemically synthesized 4-hydroxybenzoate (4HBA) is widely used in the chemical and electrical industries as a material for producing polymers such as those of the liquid crystal type. Its alkyl esters, called parabens, have been the most widely used preservatives by the food and cosmetic industries. We report here for the first time a microorganism, a marine bacterium, which biosynthesizes these petrochemical products. The marine bacterial strain, A4B-17, which was found to belong to the genus Microbulbifer on the basis of its rRNA and gyrB sequences, was isolated from an ascidian in the coastal waters of Palau. Strain A4B-17 was, surprisingly, found to produce 10 mg/liter of 4HBA, together with its butyl (24 mg/liter), heptyl (0.4 mg/liter), and nonyl (6 mg/liter) esters. We therefore characterized 23 other marine bacteria belonging to the genus Microbulbifer , which our institute had previously isolated from various marine environments, and found that these bacteria also produced 4HBA, although with low production levels (less than one-fifth of that produced by A4B-17). We also show that the alkyl esters of 4HBA produced by strain A4B-17 were effective in preventing the growth of yeasts, molds, and gram-positive bacteria.

A novel Ferrimonas species is described on the basis of phenotypic, chemotaxonomic and phylogenetic studies. Four halophilic organisms were isolated from marine sand and marine macroalgae samples by using high-pH marine agar 2216. An analysis of the nearly complete 16S rRNA gene sequences of these new isolates indicated that they were phylogenetically close (16S rRNA gene sequence similarity >99·5 %, gyrB gene sequence similarity >97·8 %), and were most closely related to Ferrimonas balearica (16S rRNA gene sequence similarity 97·1–97·3 %, gyrB gene sequence similarity 84·4–85·0 %). Chemotaxonomic data (major menaquinone MK7; major fatty acids C16 : 0 and C18 : 1 ω9c) supported the affiliation of the new isolates to the genus Ferrimonas. The results of physiological and biochemical tests allowed phenotypic differentiation of the isolates from F. balearica. It is therefore proposed that the new isolates represent a novel species with the name Ferrimonas marina sp. nov. and type strain A4D-4T (=MBIC06480T=DSM 16917T).

A carotenoid biosynthesis gene cluster mediating the production of 2-hydroxyastaxanthin was isolated from the marine bacterium Brevundimonas sp. strain SD212 by using a common crtI sequence as the probe DNA. A sequence analysis revealed this cluster to contain 12 open reading frames (ORFs), including the 7 known genes, crtW , crtY , crtI , crtB , crtE , idi , and crtZ . The individual ORFs were functionally analyzed by complementation studies using Escherichia coli that accumulated various carotenoid precursors due to the presence of other bacterial crt genes. In addition to functionally identifying the known crt genes, we found that one (ORF11, named crtG ) coded for a novel enzyme, carotenoid 2,2′-β-hydroxylase, which showed intriguingly partial homology with animal sterol-C5-desaturase. When this crtG gene was introduced into E. coli accumulating zeaxanthin and canthaxanthin, the resulting transformants produced their 2-hydroxylated and 2,2′-dihydroxylated products which were structurally novel or rare xanthophylls, as determined by their nuclear magnetic resonance and high-performance liquid chromatography/photodiode array detector/atmospheric pressure chemical ionization mass spectrometry spectral data. The new carotenoid produced was suggested to have a strong inhibitory effect on lipid peroxidation.

The two new anti-dinoflagellates, clonostachysins A and B, were obtained from a marine sponge derived fungus Clonostachys rogersoniana strain HJK9. Their chemical structures were determined by spectroscopic studies as highly N-methylated cyclic peptides of the nine amino acids. The absolute stereochemistry was elucidated by the advanced Marfey's method. Both clonostachysins A and B exhibited a selectively inhibitory effect on a dinoflagellate Prorocentrum micans at 30 microM, but had no effect on other microalgae and bacteria even at 100 microM.

A Gram-negative, strictly aerobic, lemon-yellow pigmented, non-motile, rod-shaped strain designated A7G-39(T) was isolated from a yellow coloured marine sponge (Tethya sp.). Phylogenetic analyses based on the 16S rRNA gene sequence indicated that the new strain represented a member of the family Flavobacteriaceae of the phylum Bacteroidetes and that it showed highest sequence similarity (94.1 %) to Algibacter wandonensis WS-MY22(T). The strain could be differentiated phenotypically from recognised members of the family Flavobacteriaceae. The major fatty acids of strain A7G-39(T) were identified as iso-C15:0 3-OH and summed feature 1 (iso-C15:1 H and/or C13:0 3-OH) as defined by the MIDI system. The DNA G+C content was determined to be 32.6 mol%, the major respiratory quinone was identified as menaquinone 7 (MK-7) and a polar lipid profile was present consisting of phosphatidylethanolamine, an unidentified aminolipid and an unidentified lipid. From the distinct phylogenetic position and combination of genotypic and phenotypic characteristics, the strain is considered to represent a novel genus for which the name Spongiivirga citrea gen. nov., sp. nov. is proposed. The type strain of S. citrea is A7G-39(T) (=KCTC 32990(T) = NBRC 110022(T)).

Four new bromophenols and a known bromophenol derivative have been isolated from the Palauan sponge Phyllospongia dendyi. The structures of these new compounds were established using nuclear magnetic resonance and mass spectral data. They showed antimacroalgal activity with IC50 values of 0.02–0.05 p.p.m. Moreover, all compounds were antimicroalgal against various classes of algae at concentrations of 0.5–5.0 p.p.m.

Fouling organisms cause serious problems by settling on ships’ hulls, and other marine infrastructures. Organotin compounds have been developed as effective antifoulants. However, environmental problems are associated with these compounds. It is necessary to find antifouling substances that exhibit a lower toxicity than those substances currently used. A new pyrimidine derivative, 3,4,5,6-tetrahydro-6-hydroxymethyl-3,6-dimethyl-4-pyrimidinecarboxylic acid, (compound 1) and zooanemonin (compound 2) were isolated from the marine sponge Protophlitaspongia aga as antifouling substances against the barnacle Balanus amphitrite. We also found that α-nicotinamide ribose (compound 3) inhibited germination and the attachment of Ulva spores.

The world's oceans are iron-deficient environments and there is little knowledge available regarding iron uptake by marine sponges. To understand iron-related biofunctions in marine organisms, iron-binding natural compounds from marine sponges were investigated. Here we reported a natural compound haliclonamide A and its analogue haliclonamide B were isolated from the marine sponge Haliclona sp. and their structures were investigated by spectroscopic analysis. The structure of haliclonamide A was determined to consist of novel cyclic peptides containing oxazole and methyloxazoline rings. Mass spectra revealed that these two compounds formed a 1:1 stable complex with trivalent iron but not with divalent iron. EPR analysis showed that these compounds will bind with Fe(III) and Cr(III) specifically, but will not bind to other cation ions such as Cu2+, Zn2+, Co2+, Ni2+, Al3+, and Ti3+. The binding constant of compound-iron complex was 10(19) which is lower than the binding constant of siderophores. The Fe(III) concentration in this sponge tissue was shown to be 10 and 100 times higher than the other sponge tissues and seawater. This indicated the sponge Haliclona sp. may possibly uptake iron through nonsiderophore metal-binding peptides haliclonamides A and B. It also suggests that iron uptake activity of marine organisms may occur through nonsiderophore metal-binding peptides in natural ocean.

The photooxidation of NO with oxygen over Hycom TiO2 and zeolite (A and Y form zeolite: TiO2-AZ and TiO2-YZ) composite catalysts was studied to remove NOx in the atmosphere. The photocatalytic oxidation activity of the titania in the composite catalyst in a proportion of AZ:TiO2=3:7 is about three times larger than that in the bare titania. The adsorption behaviors of NO and NO2 for the bare titania sample obey Langmuir adsorption equations of NO and NO2, respectively. In the titania–zeolite composite catalysts, the adsorption data indicate the increase in the amount of NO adsorption on the TiO2 phase and the decrease in the amount of NO2 adsorption, compared with the bare titania. The acceleration of NO photooxidation rate, resulting from the increase in the amount of NO adsorbed and the decrease in the amount of NO2 adsorbed, thus occurs on the TiO2 phase. IR spectra, when irradiating the catalysts with UV, showed the immediate formation of nitrate and NO2 species on the catalyst. The results lead to the conclusion that the zeolites promote the photocatalytic oxidation of NO over the titania.

An improved plate assay for screening antifoulants against blue mussels, Mytilus edulis galloprovincialis, was designed based on an assay contrived by Ina and colleagues. To obtain more reliable results, the number of test mussels for one sample was increased from 4 to 10. Cardboard pieces used as test plates were cut into rectangular shapes to separate the test sample and the blank zone, which are of the same area. Activity was expressed numerically. In this improved method, several inhibitors of byssus thread formation were isolated by using a smaller amount of sample per area than in the original method. Previously our laboratory developed a foot-stimulating bioassay for the same purpose. This method requires only a small amount of sample and is therefore useful for isolating antifouling substances from natural sources, including marine organisms. However, the inhibition of byssus thread formation cannot be directly evaluated in the foot-stimulating method. The improved plate assay and the foot-stimulating method were examined and compared using a secosterol from the sponge Dysidea granulosa and CuSO(4).

A new sulfoalkylresorcinol (1) was isolated from the marine-derived fungus, Zygosporium sp. KNC52. The structure of 1 was elucidated by spectroscopic methods including MS and NMR, and the absolute stereochemistry was determined by the modified Mosher's method. Compound 1 inhibited FtsZ polymerization in vitro and exhibited weak antimicrobial activity against multi-drug-resistant bacteria.

Callus of the mangrove plant, Sonneratia alba J. Smith, established from pistils of flower buds were cultured on solid Murashige and Skoog medium supplemented with 0 to 500 mM NaCl. Maximum growth was observed with 50 mM NaCl, and net growth of callus occurred for concentrations up to 200 mM NaCl. At 500 mM NaCl, growth of callus was completely inhibited, although a part of the tissue was still alive after 30 d. Cellular levels of Na+ and Cl− were greatly increased by the treatment with NaCl. Uptake of K+ was also enhanced and was accompanied by increasing levels of Na+ and Cl− so that the Na+/K+ ratio was almost constant (4.1–4.2) in callus grown with 50–200 mM NaCl. Levels of Mg2+ and Ca2+ were not changed significantly with 50–200 mM NaCl, whereas levels of free NH 4 + , NO 3 − and SO 4 2− ions, which are convertible to organic compounds, were lowest in callus grown with 50 mM NaCl. The rate of conversion of 15NH 4 + into macromolecules during 30 d culture with 0–100 mM NaCl did not vary greatly, but 200 mM NaCl reduced the biosynthesis of macromolecules from this ion. The highest rate of conversion of 15NO 3 − into macromolecules was observed at 50 mM NaCl. Identification of compatible solutes with NMR-spectroscopy indicated that mannitol is the compatible solute for intact plants of Sonneratia alba, but no accumulation of mannitol was found in calluses, not even in those grown at high concentrations of NaCl.

Two new cytotoxic antibiotics, piericidins C7 (1) and C8 (2), were isolated from a marine Streptomyces sp. High-resolution FAB-MS established the molecular formulae of 1 and 2 as C28H41NO5 and C29H43NO5, respectively. The planar structures of the piericidins were elucidated by NMR spectral analysis including COSY, HMQC and HMBC. The relative stereochemistry of 2 was determined based on NOESY and coupling constant analyses. 1 and 2 are new members of the piericidin family possessing 11,12-epoxide and 12-butenyl groups.

Novel cadmium-inducible peptides were purified from the barnacle, Megabalanus volcano. They lacked cysteine and contained high amounts of acidic residues. Their molecular weights were much smaller than those of other cadmium-inducible/binding proteins. These peptides, megabalanein A (CdIP1) and megabalanein B (CdIP2) may be the smallest metal-inducible peptides that have been found in vertebrates and invertebrates. Novel cadmium-inducible peptides, megabalanein A (a) and megabalanein B (b), were purified from the barnacle, Megabalanus volcano. Download : Download full-size image

In the present study we aimed to observe the intracellular responses when there was a hyperosmotic shock with a large shift in ionic strength in nutrient-rich and nutrient-poor external environments in order to clarify the availability of substrates. To do this, we used the halotolerant organism Brevibacterium sp. strain JCM 6894, which is able to grow in the presence of a wide range of salt concentrations. Hyperosmotic shock was induced by transferring cells in the late exponential phase of growth in a complex medium containing 0.5 M NaCl into either old or fresh culture medium containing 2 M NaCl. Changes in the growth rate, in the pH of the medium, and in the internal cation or organic solute concentrations in the cytosol after an upshock were analyzed as a function of incubation time. The cells exhibited very different responses to upshocks in fresh culture medium and in old culture medium; in fresh culture medium, growth was stimulated and the medium became more acidic, whereas the old culture medium repressed growth and the medium became more alkaline. The intracellular free Na + concentrations remained low (80 nmol mg of protein −1 ) after an upshock in fresh culture medium, although they quickly increased twofold in the old culture medium. In contrast, K + ions immediately accumulated in the cells in fresh culture medium, whereas K + ions were taken up quite slowly in old culture medium. Furthermore, the cells placed in fresh culture medium transiently accumulated alanine and glutamine in response to the upshock, but the cells placed in old culture medium did not. Growth of the Brevibacterium strain at higher levels of salinity was supported by ectoine synthesis but was not observed after the shift to high-osmolarity conditions in the old culture. In the fresh culture, however, ectoine was vigorously synthesized in cells for more than 5 h after the upshock; the concentration of ectoine in cells was more than 3,500 nmol mg of protein −1 at 10 h, which corresponded to a ninefold increase compared to the concentration before the shock. These findings are consistent with the results of an analysis of the extracellular medium composition before and after the upshock.