Contamination of water by microcystins is a global problem. These potent hepatotoxins demand constant monitoring and control methods in potable water. Promising approaches to reduce contamination risks have focused on natural microcystin biodegradation led by enzymes encoded by the mlrABCD genes. The first enzyme of this system (mlrA) linearizes microcystin structure, reducing toxicity and stability. Heterologous expression of mlrA in different microorganisms may enhance its production and activity, promote additional knowledge on the enzyme, and support feasible applications. In this context, we intended to express the mlrA gene from Sphingosinicella microcystinivorans B9 in an industrial Saccharomyces cerevisiae strain as an innovative biological alternative to degrade microcystins. The mlrA gene was codon-optimized for expression in yeast, and either expressed from a plasmid or through chromosomal integration at the URA3 locus. Recombinant and wild yeasts were cultivated in medium contaminated with microcystins, and the toxin content was analyzed during growth. Whereas no difference in microcystins content was observed in cultivation with the chromosomally integrated strain, the yeast strain hosting the mlrA expression plasmid reduced 83% of toxins within 120 h of cultivation. Our results show microcystinase A expressed by industrial yeast strains as a viable option for practical applications in water treatment.

A MeOH extract of the stem of Gmelina arborea Roxb. ex Sm. (Lamiaceae) exhibited neurite outgrowth-promoting activity in NGF-mediated PC12 cells. Bioassay-guided fractionation resulted in the isolation of eight previously undescribed prenylated coumarin compounds along with nine known compounds. Structural elucidation of these compounds was accomplished by analysis of extensive spectroscopic data, comparison with the literature, and chemical reactions. It was the first time to find prenylated coumarin compounds from G. arborea. Among the isolated compounds, N-methylflindersine and artanin showed neurite outgrowth-promoting activity in NGFmediated PC12 cells.

Five undescribed norfriedelane triterpenoids, anchietins A-E, along with three known norfriedelane triterpenoids, 21β-hydroxycaloncobalactone, and welwitschiilactones B and C, were isolated from the vine of Anchietia pyrifolia. The compounds were characterized by spectroscopic and crystallographic methods, including 2D NMR spectroscopy and single crystal X-ray crystallography. The isolated compounds were evaluated for the cytotoxic activity against HeLa and HL60 cells and for the inhibitory activity against lipopolysaccharide-induced NO production. Further, inhibitory effects on the inducible nitric oxide synthase mRNA expression levels were also assessed.

Lakes Sagami and Tsukui are reservoirs constructed by connecting to the Sagami River. Because of eutrophication of the lakes, cyanobacteria have appeared every year. This review deals with phenomena related to occurrence of cyanobacteria that have been observed for 40 years since 1974 at the lakes. These 40 years of observations raised three interesting issues including the retention of cyanobacteria on their surfaces. These phenomena have been attributed to the usual factors, such as illuminance, nutrition and water temperature, but our research results suggested that they cannot be resolved without the introduction of another factor. We have attempted to elucidate various phenomena involving cyanobacteria in lake ecosystems by chemical ecological methods using volatile organic compounds (VOCs) produced by the cyanobacteria as indicators. One of the VOCs, β-cyclocitral, was significantly involved in the above phenomena, which was considered to be produced by the carotenoid cleavage dioxygenase (CCD) of the cyanobacteria. β-Cyclocitral was not produced in the two known CCDs, but two additional CCDs to Microcystis aeruginosa participated to produce the β-cyclocitral. These CCDs did not directly produce β-cyclocitral, but it was accumulated in cells as their precursors. The released β-cyclocitral underwent a Baeyer-Villiger-like oxidation. It was speculated that Microcystis activated the CCD genes through density stress and produced β-cyclocitral, which acted as an allelopathic substance. As a result, the number of cells of cyanobacteria decreased, and the resulting nitrogen and phosphorus were fed to the living cyanobacteria. It is postulated that this "quorum sensing" was functioning in the above-mentioned issues.

A phytochemical component investigation of the bark of Ptychopetalum olacoides led to the isolation of 4 new clerodane-type diterpenoids, namely, ptycholide V ( 1 ), 7α,20-dihydroxykolavelool ( 2 ), ptycholide VI ( 3 ), and ptycholide VII ( 4 ). Their structures were elucidated by extensive spectroscopic data and comparison of NMR data with that obtained for known compounds. Keywords clerodane , , , Olacaceae , diterpenoids , ptycholide

Cyanobacteria produce numerous volatile organic compounds (VOCs) that show a lytic activity against other cyanobacteria. We found the lytic phenomenon under natural conditions and during densification experiments, and also observed the species change of the cyanobacteria during the lysis processes, in which Microcystis finally became dominant. The species change of the cyanobacteria was strongly suggested to depend on the susceptibility of the cyanobacteria toward the VOCs. To verify this suggestion, the susceptibility of the species was evaluated by the minimal inhibitory concentration (MIC) using axenic cyanobacterial strains against β-cyclocitral, its oxidation products and β-ionone with the aid of log D. It was found that the difference depended on the susceptibility of the cyanobacteria toward the VOCs, in which β-cyclocitral played a crucial role and Microcystis had a significantly protective ability compared to the other cyanobacteria. In addition, the species change of cyanobacteria was consistent with the cyanobacterial seasonal succession in Lakes Sagami and Tsukui, based on data that had been accumulated for 10 years. Conventionally, although this phenomenon could be explained by nutrient availability or the physical structure of the environment, the results of this study revealed that it was controlled by the VOCs, particularly β-cyclocitral produced by the cyanobacteria.

Os potenciais riscos associados à exposição às microcistinas (MCs) requerem o desenvolvimento de pesquisas voltadas para métodos de análise visando ao monitoramento eficiente e rápido de águas. O trabalho avaliou a aplicação dos imunoensaios ELISA desenvolvido em laboratório (LABicELISA) e o comercial (kit ELISA) para detecção de MCs em águas (n=22). O LABicELISA demonstrou ser mais sensível que o kit ELISA, apresentando limite de quantificação de 0,05 e 0,12 µg.L-1, respectivamente. Os resultados demonstraram que ambos os imunoensaios foram eficientes para detecção e análise de MCs em diferentes tipos de água (sintéticas, de mananciais destinados ao abastecimento e de reservatórios), com coeficiente de correlação de 0,997 e forte correlação na exatidão e precisão. O emprego do LABicELISA proporcionou análise com custo aproximado e reduzido de 100 vezes em relação ao do kit ELISA, podendo assim, aplicar em larga escala de abrangência para monitoramento de mananciais superficiais no país.

An abnormal growth of cyanobacteria in eutrophicated freshwaters can cause various environmental problems. In particular, Microcystis producing hepatotoxic cyclic heptapeptides microcystins (MCs) has been globally observed. Recent studies have demonstrated that matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) offers a rapid classification of cyanobacteria; however, they have not fully considered the toxicity yet. In this study, we have performed MALDI-TOF MS for intact cyanobacterial cells using Biotyper software and optimized their conditions to achieve cyanobacterial classification with the toxicity. The detection mass range used for Biotyper was extended to cover small molecules, but their intense ions were suppressed as a function of the used instrument Autoflex Speed, which enabled simultaneous observations of large molecular fingerprints and small MCs with comparable ion intensity. Hierarchical clustering of mass spectra obtained under the optimized conditions differentiated toxic and non-toxic clusters of Microcystis strains and furthermore formed a tight cluster of non-toxic strains possessing the MC biosynthesis gene mcyG. Spectral libraries were expanded to >30 genera (>80 strains) under the default and optimized conditions to improve the confidence of cyanobacterial classification. Consequently, spectral library searching allowed for characterization of cyanobacteria from a field sample as mixed toxic and non-toxic Microcystis cells, without isolating those cells.

he article The search for, and chemis.

Lakes Sagami and Tsukui are reservoirs of nearly equal size, with a short residence time and constructed by connection to the Sagami River. Because of eutrophication of the lakes, cyanobacteria appear every year. However, the occurrence of cyanobacteria in each lake was different. Therefore, in order to investigate these factors, we conducted both biotic and abiotic investigations and compared defined parameters over the past 40 years. Cyanobacteria are found in the surface layer of the lakes. In Lake Sagami, the surface layer was not largely retained and some of the cyanobacteria were found in the outflow water. When the surface layer became disturbed, Dolichospermum dominated, and Microcystis dominated when the layer stabilized due to weather effects. The outflow from Lake Tsukui was taken from a deep layer ( depth 24-32 m ) and the cyanobacteria were not affected by the outflow. Prior to the installation of the aeration systems, Microcystis dominated, and after the installation, the cyanobacteria were dispersed and Nostocales dominated, but Microcystis still dominated when the surface layer stabilized owing to the influence of the weather. As a result, the influence of nutrients and water temperature was limited and the changes in the abundance and dominant species of cyanobacteria may be influenced by the weather, the difference in the outlet structure, and ancillary facilities such as aeration systems. data, Microcystis Nostocales,

β-Cyclocitral, specifically produced by Microcystis, is one of the volatile organic compounds (VOCs) derived from cyanobacteria and has a lytic activity. It is postulated that β-cyclocitral is a key compound for regulating the occurrence of cyanobacteria and related microorganisms in an aquatic environment. β-Cyclocitral is sensitively detected when a high density of the cells is achieved from late summer to autumn. Moreover, it is expected to be involved in changes in the species composition of cyanobacteria in a lake. Although several analysis methods for β-cyclocitral have already been reported, β-cyclocitral could be detected using only solid phase micro-extraction (SPME), whereas it could not be found at all using the solvent extraction method in a previous study. In this study, we investigated why β-cyclocitral was detected using only SPME GC/MS. Particularly, three operations in SPME, i.e., extraction temperature, sample stirring rate, and the effect of salt, were examined for the production of β-cyclocitral. Among these, heating (60 °C) was critical for the β-cyclocitral formation. Furthermore, acidification with a 1-h storage was more effective than heating when comparing the obtained amounts. The present results indicated that β-cyclocitral did not exist as the intact form in cells, because it was formed by heating or acidification of the resulting intermediates during the analysis by SPME. The obtained results would be helpful to understand the formation and role of β-cyclocitral in an aquatic environment.

Nocardithiocin is a thiopeptide compound produced by the pathogenic actinomycete Nocardia pseudobrasiliensis that displays activity against Gram-positive bacteria including drug-resistant Mycobacterium tuberculosis . However, the clinical use of nocardithiocin is hampered by light sensitivity and low water solubility. To improve these properties, the sixth amino acid of core peptide was substituted for other 19 amino acids by modifying the gene sequence of precursor peptide. Ten amino acids were successfully substituted to produce 1–3 novel compounds, of which six were the expected nocardithiocin derivatives. The compounds were tested for their stability, solubility, and antimicrobial activity. Of the 17 compounds produced, ten displayed antibiotic activity and two of them improved MIC value to bacteria. Furthermore, nocardithiocin and all derivatives were stable to the light. The substitution of amino acid sequence in precursor peptide could lead the successful production of nocardithiocin derivatives and alter its property. This report represents the first step of nocardithiocin utilization, and amino acid substitution technique can be used to create desirable nocardithiocin derivatives.

Seven new vibsane-type diterpenoids, 6-O-methyl-6,7-dihydroxyvibsanin B (1), 4-hydroxyvibsanin A (2), 14(R*),15-epoxyneovibsanin B (3), 14(S*),15-epoxyneovibsanin B (4), (8Z)-neovibsanin B (5), 18-O-methylvibsanin C (6), and (8Z)-vibsanin E (7), have been isolated from the leaves of Viburnum awabuki. Their structures have been elucidated by molecular mechanics 2 (MM2) calculations and comparison of the spectroscopic data, including C-13 NMR data, with those of previously known compounds. Moreover, neovibsane-type diterpenoids 3, 4, and 5 enhanced the neurite outgrowth of NGF-mediated PC12 cells at a concentration of 40 mu M. (C) 2019 Elsevier Ltd. All rights reserved.

Sphingosinicella microcystinivorans strain B-9 has the ability to degrade cyanobacterial hepatotoxic cyclic peptides, microcystins, and nodularins. This is the first report of the complete genome sequence of the microcystin-degrading bacterium.

In this study, we confirmed the effects of light and K + concentration on the buoyancy of the cyanobacterium Microcystis aeruginosa (Kutzing) Lemmermann (NIES-843), and the relationship between the gas vesicle and buoyancy of M. aeruginosa was revealed through the culture experiment. The results showed that under laboratory conditions, light illumination and K + concentration strongly affected the flotation and settlement of M. aeruginosa , and that the floating and settling cycle of M. aeruginosa could be reproduced in a test tube by controlling light illuminance. The ability of M. aeruginosa buoyancy weakened with the increase in the K + concentration, and M. aeruginosa could not exhibit buoyancy at the K + concentration of 0.03 mol/L. The phase-contrast microscope observation revealed that M. aeruginosa that floated on the water surface had gas vesicles, while none of the gas vesicles was detected for M. aeruginosa at the bottom of a test tube.

To investigate the efficacy and safety of first-line Pazopanib advanced or metastatic renal cell carcinoma (RCC): a single-institution study. (Patients and methods) We gathered 23 RCC patients treated between April 2014 and February 2018 in our institution and examined the treatments outcome and adverse events (AEs). (Results) Their risk criteria were categorized as follows: Favorable (n=2), Intermediate (n=12) and Poor (n=9) by International mRCC Database Consortion (IMDC). Median progression free survival (PFS) was 8.6 months and median overall survival (OS) was unreached. Best response (BR) of Pazopanib was partial response (PR): n=10, 50.0%, stable disease (SD): n=8, 40.0% and progression disease (PD): n=2, 10.0% (Three patients had no evaluable data,). AEs of Grade3 had liver dysfunction (n=3), thrombocytopenia (n=2), hand-foot syndrome (n=1), vomiting (n=1) and bowel bleeding (n=1). (Conclusion) The fist-line pazopanib demonstrated good clinical benefit with well-tolerance.

SUMMARY The purpose of the present study was to demonstrate that the lysis with the blue color formation was caused by densification of the cyanobacteria, and related events of the species change in the cyanobacteria were induced by the resulting volatile organic compounds (VOCs), particularly β‐cyclocitral. In order to obtain a high cell density of cyanobacteria in the laboratory, a concentration technique (graduated cylinder method) using the buoyancy of the gas vesicles was successfully used. The collected scum contained mainly Dolichospermum spp. and Microcystis, and the dispersed cyanobacteria were concentrated in the surface layer after several hours and the concentration ratio became approximately 10. The concentrated cyanobacteria were gradually lysed, while some of the cyanobacteria sank to the bottom, which finally died and disappeared. This method has the additional advantage that it is possible to visualize the entire lysis process. During the concentration process, β‐cyclocitral and its oxidation products together with β‐ionone were significantly detected. Because β‐cyclocitral was easily oxidized to the corresponding carboxylic acid, the pH of the water in the graduated cylinder decreased to approximately 6. Under favorable conditions, lysis with the blue color from phycocyanin could be observed due to the acid stress. Overall, the results of the present study were consistent with the hypothesis that VOCs were produced when the cyanobacteria are highly dense, and that the lysis with the blue color formation occurs due to the higher density.

Strain B-9, which has a 99% similarity to Sphingosinicella microcystinivorans strain Y2, is a Gram-negative bacterium with potential for use in the degradation of microcystin-related compounds and nodularin. We attempted to extend the application area of strain B-9 and applied it to mycotoxins produced by fungi. Among the tested mycotoxins, only ochratoxin A was completely hydrolyzed to provide the constituents ochratoxin α and l-phenylalanine, and levels of fumonisin B1 gradually decreased after 96 h. However, although drugs including antibiotics released into the aquatic environment were applied for microbial degradation using strain B-9, no degradation occurred. These results suggest that strain B-9 can only degrade amino acid-containing compounds. As expected, the tested compounds with amide and ester bonds, such as 3,4-dimethyl hippuric acid and 4-benzyl aspartate, were readily hydrolyzed by strain B-9, although the sulfonamides remained unchanged. The ester compounds were characteristically and rapidly hydrolyzed as soon as they came into contact with strain B-9. Furthermore, the degradation of amide and ester compounds with amino acids was not inhibited by the addition of ethylenediaminetetraacetic acid (EDTA), indicating that the responsible enzyme was not MlrC. These results suggest that strain B-9 possesses an additional hydrolytic enzyme that should be designated as MlrE, as well as an esterase.

To develop the Javanese ginger Bangle (Zingiber purpureum), which contains neurotrophic phenylbutenoid compounds named banglenes 1 and 2, as a functional food additive for protection of neurodegenerative disease, we investigated conversion method from 3 to 1 and 2 under practical conditions available in food processing steps. After several trials, we found that dimerization of 3 proceeded smoothly under refluxing EtOH/H2O (1/3) in the presence of ubiquinone (0.1 eq) in sealed tube, giving rise to 1 and 2 in 54% yield. When this condition was applied for 70% EtOH extract of bangle, content of 1 and 2 in the extract was increased 2-fold by 2.1%.