Phospholipid fatty acid composition of gorgonians of the genus Pseudopterogorgia: Identification of tetracosapolyenoic acids

The phospholipid fatty acid composition of the Caribbean gorgonians Pseudopterogorgia acerosa (Pallas), Pseudopterogorgia americana (Gmelin), Pseudopterogorgia bipinnata (Verrill) and Pseudopterogorgia rigida (Bielschowsky) is described for the first time. The main phospholipids identified were phosphatidylethanolamine, phosphatidylcholine and phosphatidylserine. All four gorgonians presented a similar phospholipid fatty acid composition. The main fatty acids were 16:0, 18:3(n-6), 18:4(n-3), 20:4(n-6), 22:6(n-3), 24:5(n-6) and 24:6(n-3). In all of the studied Pseudopterogorgia gorgonians, high amounts of the tetracosapolyenoic fatty acids 24:5(n-6) and 24:6(n-3) were identified. In the four gorgonians studied, n-6 polyunsaturated fatty acids predominated. These results suggest that the occurrence of tetracosapolyenoic fatty acids in the Gorgoniidae is more general than previously recognized.     

Nonlinkage between C6 and chromosome 6 markers

Summary C6 typing was performed in a family material by two different techniques: serum or plasma samples were subjected either to high-voltage agarose gel electrophoresis or to isoelectric focusing in polyacrylamide gel slabs. Proteins with C6 activity were then visualized by a specific, hemolytic assay.In 81 unrelated adults within the family material the following allele frequencies were found: C6 ^A:0.61 and C6 ^B:0.39.Linkage studies exclude linkage between C6 and HLA region marker loci, and also between C6 and another chromosome 6 marker locus PGM3.     

Interaction of Sesamin and Eicosapentaenoic Acid against Δ5 Desaturation and n-6/ n-3 Ratio of Essential Fatty Acids in Rat

Sesamin is a specific inhibitor of Δ5 desaturation, the conversion from dihomo-γ-linolenic acid (20: 3, n-6) to arachidonic acid (AA, 20: 4, n-6). Previously, we reported that sesamin inhibited Δ5 desaturation of n-6 fatty acids in rat hepatocytes but not that of n-3 fatty acids, from 20: 4 (n-3) to eicosapentaenoic acid (EPA, 20: 5, n-3). In this study, we investigated the interaction of sesamin and EPA on Δ5 desaturation of both series and the n-6/n-3 fatty acids ratio by measuring actural fatty acid contents in vivo. Rats were fed three types of dietary oils; 1) linoleic acid (LA, 18: 2, n-6): linolenic acid (LLA, 18: 3, n-3) = 3: 1, n-6/n-3 ratio of 3: 1 (LA group), 2) LA: LLA =1: 3, n-6/n-3 ratio of 1: 3 (LLA group), 3) LA: LLA: EPA =1: 0.5: 3, n-6/n-3 ratio of 1: 3.5 (EPA group) with or without sesamin (0.5% w/w) for 4 weeks. In all groups, sesamin administration increased the content of dihomo-γ-linolenic acid (20: 3, n-6) in the liver and decreased the Δ5 desaturation index of n-6 fatty acid, the ratio of 20: 4/20: 3 (n-6). On the contrary, the Δ5 desaturation index of n-3 fatty acid, the ratio of 20: 5 + 22: 5 + 22: 6/20: 4 (n-3), was increased by the administration of sesamin. These results suggest that sesamin inhibits the A5 desaturation of n-6 fatty acid, but not that of n-3 fatty acid in rat livers. Sesamin administration decreased incorporation of EPA (n-3) and simultaneously increased the AA (n-6) content in the liver. The n-6/n-3 ratio in the liver was increased by administering sesamin under n-3 rich conditions, i.e., the LLA and EPA groups.     

Cloning, expression and function of phosphate transporter encoded gene in Oryza sativa L.

OsPT6:1, a phosphate transporter encoding gene from the leaf samples of Oryza sativa, was identified through PCR with specifically designed primers. The phylogenetic analysis and the conserved amino acid residue site detection suggested OsPT6:1 a possible high-affinity phosphate transporter encoding gene. In situ hybridization and RT-PCR demonstrated the expression of OsPT6:1 in both roots and leaves. The peak expression signal was observed in mesophyll cells under low phosphorus (P) induction. A homologous recombination study indicated that OsPT6:1 can enhance the Pi uptake efficiency of Pichia pastoris. At the meantime, the introduction of OsPT6:1 was able to complement the Pi uptake function of yeast cells with high-affinity phosphate transporters deficient. Those results substantiated our contention that OsPT6:1 encoded a high-affinity phosphate transporter of Oryza sativa.     

Cloning, expression and function of phosphate transporter encoded gene in Oryza sativa L.

OsPT6:1, a phosphate transporter encoding gene from the leaf samples of Oryza sativa, was identified through PCR with specifically designed primers. The phylogenetic analysis and the conserved amino acid residue site detection suggested OsPT6:1 a possible high-affinity phosphate transporter encoding gene. In situ hybridization and RT-PCR demonstrated the expression of OsPT6:1 in both roots and leaves. The peak expression signal was observed in mesophyll cells under low phosphorus (P) induction. A homologous recombination study indicated that OsPT6:1 can enhance the Pi uptake efficiency of Pichia pastoris. At the meantime, the introduction of OsPT6:1 was able to complement the Pi uptake function of yeast cells with high-affinity phosphate transporters deficient. Those results substantiated our contention that OsPT6:1 encoded a high-affinity phosphate transporter of Oryza sativa. These authors contributed equally to this work.     

Substrate specificities of lipases in view of kinetic resolution of unsaturated fatty acids

Several commercially available lipases have been evaluated with regard to their substrate specificity in the esterification of fatty acids having specific positions of cis double bonds, e.g. petroselinic acid (n-12 18:1), alpha-linolenic acid (n-3 18:3), gamma-linolenic acid (n-6 18:3), stearidonic acid (n-3 18:4), dihomogamma-linolenic acid (n-6 20:3), eicosapentaenoic acid (n-3 20:5) and docosahexaenoic acid (n-3 22:6), with n-butanol. A common feature of most lipases, e.g. those from Penicillium cyclopium, Candida cylindracea, Mucor miehei, Rhizopus arrhizus and Penicillium sp. is that fatty acids having the first double bond from the carboxyl end as a cis-4 (n-3 22:6), cis-6 (n-12 18:1, n-6 18:3, n-3 18:4) or a cis-8 (n-6 20:3) double bond are strongly discriminated against compared to the other fatty acids, such as myristic acid (14:0), the reference standard, and n-3 18:3. In the case of the lipase from porcine pancreas, however, the discrimination against the above fatty acids is not as strong as with the other lipases. In contrast, the lipase from Chromobacterium viscosum shows a preference for n-12 18:1, n-6 18:3 and n-3 18:4. The observed substrate specificities can be utilized for enrichment of particular fatty acids by lipase-catalysed kinetic resolution from fatty acid mixtures, derived from naturally occurring fats and other lipids.Dedicated to Prof. David A. Walker, Robert Hill Institute, Department of Animal and Plant Sciences, University of Sheffield, Sheffield, UK, on the occasion of his sixty-fifth birthday on 18 August 1993 Correspondence to: K. D. Mukherjee     

Isolation of Stable Hemolysin and Catalase Variants of Staphylococcus aureus S6C Includes One with an Exoprotein-Deficient Phenotype

Previous studies suggested that the exoprotein-deficient phenotype of a Δ1058::Tn551 insertion/deletion mutant of Staphylococcus aureus S6C was not owing to the insertion/deletion event, but instead was owing to the inherent instability of the agrC gene during transduction of the Δ1058::Tn551 region into S6C. The purpose of the following study was to examine S6C as a potential source of exoprotein-deficient mutants that would account for their appearance after transposition and transduction. Four stable variants of S6C were isolated that differed in their hemolysin and catalase activities. Surprisingly, the agr regulatory molecule, RNAIII, was undetectable in one of these variants, which most likely accounted for the exoprotein-deficient phenotype of this variant. When the original Δ1058::Tn551 mutation was transduced into the hemolytic, catalase-positive variant of S6C, none of the transductants exhibited an exoprotein-deficient phenotype. These data suggest that, while the exoprotein-deficient phenotype of the S6C variant is most likely due to mutations in the agr regulatory system, these mutations are not caused by the transduction of the Δ1058::Tn551 region into S6C, but instead already exist in an exoprotein-deficient variant of S6C. Received: 6 November 2000 / Accepted: 16 January 2001     

Cloning and characterization of two cytochrome P450 CYP6AX1 and CYP6AY1 cDNAs from Nilaparvata lugens Stål (Homoptera: Delphacidae)

Two full‐length P450 cDNAs, CYP6AX1 and CYP6AY1, were cloned from the brown planthopper Nilaparvata lugens Stål (Homoptera: Delphacidae). Both CYP6AX1 and CYP6AY1 are typical microsomal P450s and their deduced amino acid sequences share common characteristics with other members of the insect P450 CYP6 family. CYP6AX1 and CYP6AY1 show the highest percent identity (36%) of amino acid to each other; both of them have 31–33% amino acid identity with CYP6B1 from Papilio polyxenes (Lepidoptera: Papilionidae), CYP6B4 from Papilio glaucus (Lepidoptera: Papilionidae), and CYP6B8 from Helicoverpa zea (Lepidoptera: Noctuidae). Phylogenetic analysis showed the clustering of CYP6AX1 and CYP6AY1 was in the clade including CYP6AE1 from Depressaria pastinacella (Lepidoptera: Oecophoridae) and the CYP6B family members from Helicoverpa and Papilio species. Northern blot analysis revealed that both of the P450s were induced by the resistant rice variety B5 (Oryza sativa L), and CYP6AY1 was expressed at a higher level than CYP6AX1. The results suggest that more than one P450s are likely involved in metabolism of rice allelochemicals and that they are possibly important components in adaptation of Nilaparvata lugens to host rice. Arch. Insect Biochem. Physiol. 64:88–99, 2007. © 2007 Wiley‐Liss, Inc.     

New species of the subfamily Conoderinae (Coleoptera: Curculionidae) in Dominican amber

Eulechriops argyrosoman. sp. (Curculionidae: Conoderinae: Lechropini) and Geratozygops platysoman. sp., Geratozygops stenosoman. sp. and Geratozygops arsinotusn. sp. (Curculionidae: Conoderinae: Zygopini) are described from Dominican amber. The small size, nearly parallel sides of the pronotum and silvery sheen distinguish Eulechriops argyrosoma from extant members of the genus, which have not been recorded from Hispaniola. Size and rostral and pronotal characters separate the three Geratozygops species from the single species (Geratozygops atropos) previously described from Dominican amber.http://www.zoobank.org/urn:lsid:zoobank.org:pub:89BC7F43-6C1A-4A6A-839B-CE94C7F6F140     

Biosynthesis of polyunsaturated fatty acids in zooxanthellae and polyps of corals

The fatty acid (FA) composition of zooxanthellae, polyp tissue, and intact colonies was determined in soft coral Sinularia sp. and hard coral Acropora sp. Analysis of the distribution of polyunsaturated fatty acids (PUFAs) among the zooxanthellae and the host organism showed that 18: 3n-6 and C_18–22 PUFAs of the n-3 series (18: 4n-3, 20: 5n-3, 22: 5n-3, and 22: 6n-3) were mainly synthesized by the zooxanthellae and that C_20–22 PUFAs of the n-6 series (20: 3n-6, 20: 4n-6, and 22: 4n-6) were synthesized in the polyp tissue. Soft coral polyps were able to synthesize tetracosapolyenoic FAs (24: 5n-6 and 24: 6n-3) and 18: 2n-7, their zooxanthellae synthesized C₁₆ PUFAs (16: 2n-7, 16: 3n-4, and 16: 4n-1). It is supposed that the biosynthesis of 16: 2n-7 in Sinularia sp. and 18: 3n-6 in Acropora sp. is catalyzed by Δ6 desaturase. The relatively even distribution of three FAs (18: 2n-6, 18: 3n-6, and 16: 2n-7) among lipids of zooxanthellae and coral polyps indicates the possible transport of these FAs between symbionts and the host organism.     

Twelve newSalmonella types

Twelve hitherto unrecognizedSalmonella types are described. S. heerlen (11 : i : 1,6) andS. sloterdijk (1, 4, 12, 27 : z 35 : z 6) were isolated from patients in the Netherlands,S. maartensdijk (40 : g, p) was isolated from a healthy calf,S. maastricht (11 : z 41 : 1, 2(7)) from imported fishmeal,S. parera (11 : z 4, z 23 : -.) from water on the island Bonaire,S. putten (13, 23, 36 : d : 1, w),S. hoograven (50 : z 10 : z 6 : z 42),S. schalkwijk ((6), 14, (24) : i : e, n, ...),S. hilversum (30 : k : 1, 2) andS. harmelen (51 : z 4, z 23 : -.) from reptiles,S. breukelen (6, 8 : 1, z 13 : e, n, z 15) from a cuscus andS. maarssen (9, 46 : z 4, z 24 : z 39 : z 42) from a lizard.     

Two new species of Parapentaneura (Diptera: Chironomidae: Tanypodinae) from Brazil,with keys to the males and immature stages

Two new species of Parapentaneura Stur, Fittkau et Serrano, 2006 (Diptera: Chironomidae: Tanypodinae) from southeastern Brazil are described and figured as male, pupa and larva: P. brunnescens and P. flavescens. Keys to the males, pupae and larvae of known species of Parapentaneura are provided.

http://www.zoobank.org/urn:lsid:zoobank.org:pub:E6860EBE-B781-4DDA-9650-6E6EA3A0309E     

Occurrence of Melibiose‐Containing Glycosphingolipids in a Sample of a Sponge‐Coral Association (Desmapsamma anchorata/Carijoa riisei)

In our research on biologically active compounds from Vietnamese marine invertebrates, rare melibiose‐containing glycosphingolipids were found in a sample of a sponge‐coral association (Desmapsamma anchorata/Carijoa riisei). Melibiosylceramides were analyzed as constituents of some multi‐component RP‐HPLC fractions, and the structures of 14 new ( 1b , 3b , 4a – 4c , 6a – 6c , 8b , 9a , 9b , 10b , 11a , 11b ) and five known ( 2b , 5a – 5c , 7b ) natural compounds were elucidated using NMR, mass spectrometry, optical rotation, and chemical transformations. These α‐d ‐Galp‐(1→6)‐β‐d ‐Glcp‐(1 1)‐ceramides (presumably sponge‐derived compounds) were shown to contain phytosphingosine‐type n‐t17:0 ( 1 ), (6E)‐n‐t17:1 ( 2 ), i‐t17:0 ( 3 ), n‐t18:0 ( 4 ), (6E)‐n‐t18:1 ( 5 ), i‐t18:0 ( 6 ), (6E)‐i‐t18:1 ( 7 ), i‐t19:0 ( 8 ), (6E)‐i‐t19:1 ( 9 ), ai‐t19:0 ( 10 ), and (6E)‐ai‐t19:1 ( 11 ) backbones N‐acylated with saturated straight‐chain (2R)‐2‐hydroxy C₂₁ ( a ), C₂₂ ( b ), and C₂₃ ( c ) acids. Characteristic trends in the fragmentations of the terminal parts of tetraacetylated normal‐chain and iso‐ and anteiso‐branched sphingoid bases were observed using GC/MS. The total sum of melibiosylceramides and compound 5b caused a reduction in colony formation of human melanoma cells.     

Inhibition of TRAF6 ubiquitin-ligase activity by PRDX1 leads to inhibition of NFKB activation and autophagy activation

TRAF6 (TNF receptor associated factor 6) plays a pivotal role in NFKB activation and macroautphagy/autophagy activation induced by TLR4 (toll like receptor 4) signaling. The objective of this study was to determine the functional role of PRDX1 (peroxiredoxin 1) in NFKB activation and autophagy activation. PRDX1 interacted with the ring finger domain of TRAF6 and inhibited its ubiquitin-ligase activity. The inhibition on TRAF6 ubiquitin-ligase activity by PRDX1 induced the suppression of ubiquitination of an evolutionarily conserved signaling intermediate in Toll pathways (ECSIT) essential for NFKB activation and BECN1 (beclin 1) required for autophagy activation. An inhibitory effect of PRDX1 on TRAF6 was clearly evidenced in PRDX1-knockdown (PRDX1KD) THP-1, PRDX1KD MDA-MB-231, and PRDX1KD SK-HEP-1 cells. PRDX1KD THP-1 cells showed increases of NFKB activation, pro-inflammatory cytokine production, NFKB-dependent gene expression induced by TLR4 stimulation, and resistance against Salmonella typhimurium infection. Additionally, migration and invasion abilities of PRDX1KD MDA-MB-231 and PRDX1KD SK-HEP-1 cancer cells were significantly enhanced compared to those of control cancer cells. Taken together, these results suggest that PRDX1 negatively regulates TLR4 signaling for NFKB activation and autophagy functions such as bactericidal activity, cancer cell migration, and cancer cell invasion by inhibiting TRAF6 ubiquitin-ligase activity.

Abbreviations: 3-MA: 3-methyladenine; BECN1: beclin 1; CHUK/IKKA: conserved helix-loop-helix ubiquitous kinase; ECSIT: ECSIT signalling integrator; ELISA: enzyme-linked immunosorbent assay; NFKB: nuclear factor kappa-light-chain-enhancer of activated B cells; IB: immunoblotting; IKBKB/IKKB: inhibitor of nuclear factor kappa B kinase subunit beta; IL1B: interleukin 1 beta; IL6: interleukin 6; IP: immunoprecipitation; LPS: lipopolysaccharide; MAP1LC3/LC3: microtuble associated protein 1 light chain 3; MAP3K7/TAK1: mitogen-activated protein kinase kinase kinase 7; MAPK14/p38: mitogen-activated protein kinase 14; mROS: mitochondrial reactive oxygen species; PRDX1: peroxiredoxin 1; PRDX6: peroxiredoxin 6; RELA/p65: RELA proto-oncogene, NF-kB subunit; TRAF6 TNF: receptor associated factor 6.     

Tyrosylprotein sulfotransferase regulates collagen secretion in Caenorhabditis elegans

The sulfation of tyrosine residues is an important post-translational modification involved in the regulation of protein function. We examined the activity of worm tyrosylprotein sulfotransferase (TPST-1) on a typical cuticle collagen, ROL-6, in C. elegans. We verified that TPST-1 sulfates three tyrosine residues of ROL-6 in vitro. We found that these tyrosine residues are important for the secretion of ROL-6::GFP. Mutant ROL-6::GFP proteins that contain more than two substitutions of the target tyrosine residues are severely deficient in cuticle localization. Consistently, knock down of tpst-1 blocked the cuticle localization of ROL-6::GFP. Therefore, the sulfation of ROL-6 by TPST-1 is critical for the proper localization of ROL-6. We also confirmed that worm TPST-1 is localized to the trans-Golgi network (TGN). Our results indicate that TPST-1 regulates cuticle organization by promoting the transport of ROL-6 from the TGN to the cuticle.     

Functional characterization of a delta 6-desaturase gene from the black seabream (<Emphasis Type="Italic">Acanthopagrus schlegeli</Emphasis>)

Delta 6-fatty acid desaturase (D6DES) is used in the synthesis of polyunsaturated fatty acids (PUFAs) from microorganisms to higher animals, including arachidonic acid (ARA) and eicosapentaenoic acid (EPA). A 1,338 bp full-length cDNA encoding D6DES was cloned from Acanthopagrus schlegeli (AsD6DES) through degenerate- and RACE-PCR methods. A recombinant vector expressing AsD6DES (pYES-AsD6DES) was subsequently constructed and transformed into Saccharomyces cerevisiae to test the enzymatic activity of AsD6DES towards the production of n-6 and n-3 fatty acids. The exogenously expressed AsD6DES produced γ-linolenic acid (18:3 n-6) and stearidonic acid (18:4n-3) at 26 and 36% from exogenous linoleic acid (18:2n-6) and α-linolenic acid (18:3n-3), respectively, indicating that it is essentially a delta 6-fatty acid desaturase.     

HIGH POLYUNSATURATED FATTY ACID LEVELS IN TWO SUBTROPICAL MACROALGAE,CLADOSIPHON OKAMURANUS AND CAULERPA LENTILLIFERA1

The lipid and fatty acid compositions in two edible subtropical algae (the brown alga Cladosiphon okamuranus Tokida and the green alga Caulerpa lentillifera J. Agardh) were determined to clarify their lipid characteristics and nutritional values. Glycolipids and phospholipids were the major lipid classes, with significant levels of triacylglycerols. Polyunsaturated fatty acids (PUFA) were the major fatty acids of both algae. The lipid class composition and major fatty acids were similar in both the algal species, irrespective of wild and cultured specimens. Typical n‐6 PUFA, such as 18:2n‐6 (linoleic acid) and 20:4n‐6 (arachidonic acid), occurred in characteristically high levels in both of the algae. High levels of n‐3 PUFA were measured in all lipid classes of both species without 22:6n‐3 (docosahexaenoic acid), 18:3n‐3, 18:4n‐3, and 20:5n‐3 (eicosapentaenoic acid) for Cl. okamuranus; and 16:3n‐3, 18:3n‐3, and 20:5n‐3 for Ca. lentillifera. The finding suggests that the green algal species, which mainly biosynthesizes short‐chain (C₁₆ and C₁₈) PUFA, differs from that of the brown alga, which is capable of biosynthesizing high 20:5n‐3 levels. The PUFA levels in glycolipids of the two algal species comprised up to 60%, even though they are subtropical marine species. High n‐6 PUFA levels in the algal lipids probably influence the significant levels of n‐6 PUFA in herbivorous fishes, because the n‐6 PUFA levels in marine fish lipids are generally undetectable or negligible.     

Establishment of highly efficient transgenic system for black soldier fly (Hermetia illucens)

The black soldier fly (BSF), Hermetia illucens, is a promising insect for mitigating solid waste problems as its larvae are able to bioconvert organic waste into valuable biomass. We recently reported a high-quality genome assembly of the BSF; analysis of this genome sequence will further the understanding of insect biology and identify genes that can be manipulated to improve efficiency of bioconversion. To enable genetic manipulation of the BSF, we have established the first transgenic methods for this economically important insect. We cloned and identified the ubiquitous actin5C promoter (Hiactin5C-p3k) and 3 endogenous U6 promoters (HiU6:1, HiU6:2, and HiU6:3). The Hiactin5C promoter was used to drive expression of a hyperactive variant of the piggyBac transposase, which exhibited up to 6-fold improvement in transformation rate when compared to the wild-type transposase. Furthermore, we evaluated the 3 HiU6 promoters using this transgenic system. HiU6:1 and HiU6:2 promoters provided the highest knockdown efficiency with RNAi and are thus promising candidates for future Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) development. Overall, our findings provide valuable genetic engineering toolkits for basic research and genetic manipulation of the BSF.     

Comparing formulations for a mixed-microbial biopesticide with Bacillus thuringiensis var. kurstaki and Beauveria bassiana blastospores

Developing a dual agent biopesticide formulation made with blastospores of Beauveria bassiana (Balsamo) Vuillemin strain GHA (Bb) at the ratio of 1:1 mixed with traditional fermentation production of Bacillus thuringiensis kurstaki (Btk) spores and crystals were studied. Twelve potential formulation ingredients were screened for their impact on insecticidal activity, microbe viability, UV protection and product suspensibility of a spray dried Bt/Bb (1:1) mixture applications to cabbage plants against cabbage looper, Trichoplusia ni. Based on the results, Spray-dried mixtures of Bt/Bb made with biochar, chitosan and skim milk powder formulations gave the greatest efficacy and higher solar stability when exposed to simulated sunlight. They therefore can be used as a formulation that combines Bt with Bb in one product. However, the formulation made with aitkin component had significantly more colony forming units (CFUs) of Bt, whereas, formulations made with albumin, sodium carbonate and pharmamedia had more Bb CFUs than other formulations. Albumin, pharmamedia, skim milk powder and biochar had significantly better suspension in water (less settling) than others. These results support the concept of formulating the two biocontrol agents, B. thuringiensis and B. bassiana blastospores for greater efficacy, more viability and longer field persistence as an environmentally friendly sustainable pest control tool.

LSID:urn:lsid:zoobank.org:pub:568D6FE6-E876-417B-BE39-C51F7CBAD6AB