Research Article

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Insect bioactive capabilities of Epichloë festucae var lolii AR48 infected Lolium perenne : a thesis presented in partial fulfilment of the requirements for the degree of Doctor of Philosophy in Biochemistry at Massey University, Manawatū, New Zealand

Figures printed with permission from respective publishers.As the modern world expands and develops, new innovative methodologies for more efficient and environmentally friendly agricultural practices are required. Loss of crops through abiotic (e.g. drought) and biotic (e.g. herbivory) stresses has a major effect on the success of an agricultural industry. For animal production pasture crops are a key aspect of animal husbandry and directly affects yield and health. Symbiotic fungi belonging to the genus Epichloë form associations with cool season forage grasses and have been exploited as a new innovative method for insect pest management. Ryegrass infected with the asexual E. festucae var lolii strain AR48 has insect bioactivity against both the stem boring fly (SBF-Ceradontha australis) and cutworm moth caterpillar (CC -Agrotis ipsilion). The bioactive/s targeting both insects is currently unknown. The aim of this thesis was to identify the gene/s and/or bioactive/s present in AR48 infected ryegrass that have bioactivity against the SBF and/or CC. Two approaches were taken; the known insect bioactive secondary metabolite pathways in Epichloë were investigated in AR48 through bioinformatics and mass spectrometry, and the gene ‘makes caterpillars floppy’ (mcf), encoding an insect toxin like protein, was investigated through reverse genetics and insect bioactivity trials. A new indole diterpene compound (IDT) was identified in AR48 infected plant material and this compound was absent in other Epichloë strains that do not have SBF and CC bioactivity. The same mcf gene allele as that present in the E. typhina mcf model, previously identified as having CC bioactivity, is present and predicted to be functional in AR48. The other Epichloë strains also have mcf genes predicted to be functional, however the mcf allele is different to the bioactive E. typhina mcf model. Overall, this project was able to identify a new IDT compound with potential insect bioactivity as well as identify two Epichloë mcf gene alleles that potentially have differing insect bioactivities

Comparative Analysis Of Zebrafish And Planarian Model Systems For Developmental Neurotoxicity Screens Using An 87-Compound Library

There is a clear need to establish and validate new methodologies to more quickly and efficiently screen chemicals for potential toxic effects, particularly on development. The emergence of alternative animal systems for rapid toxicology screens presents valuable opportunities to evaluate how systems complement each other. In this article, we compare a chemical library of 87-compounds in two such systems, developing zebrafish and freshwater planarians, by screening for developmental neurotoxic effects. We show that the systems’ toxicological profiles are complementary to each other, with zebrafish yielding more detailed morphological endpoints and planarians more behavioral endpoints. Overall, zebrafish was more sensitive to this chemical library, yielding 86/87 hits, compared to 50/87 hits in planarians. The difference in sensitivity could not be attributed to molecular weight, Log Kow or the bioconcentration factor. Of the 87 chemicals, 28 had previously been evaluated in mammalian developmental neuro- (DNT), neuro- or developmental toxicity studies. Of the 28, 20 were hits in the planarian, and 27 were hits in zebrafish. Eighteen of the 28 had previously been identified as DNT hits in mammals and were highly associated with activity in zebrafish and planarian behavioral assays in this study. Only 1 chemical (out of 28) was a false negative in both zebrafish and planarian systems. Differences in endpoint coverage and system sensitivity illustrate the value of a dual systems approach to rapidly query a large chemical-bioactivity space and provide weight-of-evidence for prioritization of chemicals for further testing

Bioactivity and structural properties of chimeric analogs of the starfish SALMFamide neuropeptides S1 and S2

The starfish SALMFamide neuropeptides S1 (GFNSALMFamide) and S2 (SGPYSFNSGLTFamide) are the prototypical members of a family of neuropeptides that act as muscle relaxants in echinoderms. Comparison of the bioactivity of S1 and S2 as muscle relaxants has revealed that S2 is ten times more potent than S1. Here we investigated a structural basis for this difference in potency by comparing the bioactivity and solution conformations (using NMR and CD spectroscopy) of S1 and S2 with three chimeric analogs of these peptides. A peptide comprising S1 with the addition of S2's N-terminal tetrapeptide (Long S1 or LS1; SGPYGFNSALMFamide) was not significantly different to S1 in its bioactivity and did not exhibit concentration-dependent structuring seen with S2. An analog of S1with its penultimate residue substituted from S2 (S1(T); GFNSALTFamide) exhibited S1-like bioactivity and structure. However, an analog of S2 with its penultimate residue substituted from S1 (S2(M); SGPYSFNSGLMFamide) exhibited loss of S2-type bioactivity and structural properties. Collectively, our data indicate that the C-terminal regions of S1 and S2 are the key determinants of their differing bioactivity. However, the N-terminal region of S2 may influence its bioactivity by conferring structural stability in solution. Thus, analysis of chimeric SALMFamides has revealed how neuropeptide bioactivity is determined by a complex interplay of sequence and conformation

Selection of gonadotrophin surge attenuating factor phage antibodies by bioassay

Peer reviewedPublisher PD

Моделювання рецептур сироватково-полуничних напоїв профілактичного призначення

Expediency of the development of formulae and innovative technologies for production of prophylactic application drinks possessing antioxidant, probiotic and hepatoprotective properties with the use of the secondary dairy product – whey, as well as the domestic vegetable raw materials having a high content of bioactive substances has been substantiated. Formulation composition of the prophylactic drinks based on cheese whey, extract of Tagetes patula flowers and the berry filler “Strawberry” with the use of the response surface method has been developed. Bioactivity of the drinks and the complex quality indicator which accounts for the total influence of the bioactivity, organoleptic assessment and weight coefficients of the specified unit indicators were taken as the optimization criteria; as the independent factors that were varied in the course of the experiment, the mass fractions of the marigold flowers extract and the strawberries filler were selected. It is recommended that the mass fractions of the berry filler “Strawberry” and the extract of Tagetes patula flowers in the prophylactic drinks are set as 7 and 20 % of the finished product, accordingly. The practical mass fraction of the citric acid of 0.2 % was determined as it ensures high organoleptic characteristics of the finished drinks. Recommendations are given concerning development of innovative technologies of unfermented and fermented strawberry whey drinks of prophylactic application enriched with the extract of Tagetes patula flowers.Обґрунтовано доцільність розроблення рецептур та інноваційних технологій напоїв профілактичного призначення з антиоксидантними, пробіотичними та гепатопротекторними властивостями з використанням вторинної молочної сировини – сироватки, а також вітчизняної сировини рослинного походження з високим вмістом біологічно активних речовин. Розроблено рецептурний склад напоїв профілактичного призначення на основі сирної сироватки, екстракту квітів Tagetes patula і ягідного наповнювача «Полуниця» з використанням методології поверхні відгуку. У якості критеріїв оптимізації обрано біологічну активність напоїв і комплексний показник якості, який враховує сукупний вплив біологічної активності, органолептичної оцінки і коефіцієнтів вагомості зазначених одиничних показників; незалежними факторами, які варіювалися, в експерименті – масові частки екстракту квітів чорнобривців і полуничного наповнювача. Рекомендовано масову частку ягідного наповнювача «Полуниця» та екстракту квітів Tagetes patula в напоях профілактичного призначення встановити 7 і 20 % від маси готового продукту відповідно. Визначено раціональну масову частку лимонної кислоти – 0,2 %, яка забезпечує високі органолептичні характеристики готових напоїв. Наведено рекомендації з розробки інноваційних технологій неферментованих і ферментованих сироватково-полуничних напоїв профілактичного призначення, збагачених екстрактом квітів Tagetes patula

Surface Modification of Biodegradable Porous Mg Bone Scaffold Using Polycaprolactone/Bioactive Glass Composite

A reduction in the degradation rate of magnesium (Mg) and its alloys is in high demand to enable these materials to be used in orthopedic applications. For this purpose, in this paper, a biocompatible polymeric layer reinforced with a bioactive ceramic made of polycaprolactone (PCL) and bioactive glass (BG) was applied on the surface of Mg scaffolds using dip-coating technique under low vacuum. The results indicated that the PCL-BG coated Mg scaffolds exhibited noticeably enhanced bioactivity compared to the uncoated scaffold. Moreover, the mechanical integrity of the Mg scaffolds was improved using the PCL-BG coating on the surface. The stable barrier property of the coatings effectively delayed the degradation activity of Mg scaffold substrates. Moreover, the coatings induced the formation of apatite layer on their surface after immersion in the SBF, which can enhance the biological bone in-growth and block the microcracks and pore channels in the coatings, thus prolonging their protective effect. Furthermore, it was shown that a three times increase in the concentration of PCL-BG noticeably improved the characteristics of scaffolds including their degradation resistance and mechanical stability. Since bioactivity, degradation resistance and mechanical integrity of a bone substitute are the key factors for repairing and healing fractured bones, we suggest that PCL-BG is a suitable coating material for surface modification of Mg scaffolds