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

CaSiO3 microstructure modulating the in vitro and in vivo bioactivity of poly(lactide-co-glycolide) microspheres

Poly (lactide-co-glycolide) (PLGA) microspheres have been used for regenerative medicine due to their ability for drug delivery and generally good biocompatibility, but they lack adequate bioactivity for bone repair application. CaSiO3 (CS) has been proposed as a new class of material suitable for bone tissue repair due to its excellent bioactivity. In this study, we set out to incorporate CS into PLGA microspheres to investigate how the phase structure (amorphous and crystal) of CS influences the in vitro and in vivo bioactivity of the composite microspheres, with a view to the application for bone regeneration. X-ray diffraction (XRD), N2 adsorption-desorption analysis and scanning electron microscopy (SEM) were used to analyze the phase structure, surface area/pore volume, and microstructure of amorphous CS (aCS) and crystal CS (cCS), as well as their composite microspheres. The in vitro bioactivity of aCS and cCS – PLGA microspheres was evaluated by investigating their apatite-mineralization ability in simulated body fluids (SBF) and the viability of human bone mesenchymal stem cells (BMSCs). The in vivo bioactivity was investigated by measuring their de novo bone-formation ability. The results showed that the incorporation of both aCS and cCS enhanced the in vitro and in vivo bioactivity of PLGA microspheres. cCS/PLGA microspheres improved better in vitro BMSC viability and de novo bone-formation ability in vivo, compared to aCS/PLGA microspheres. Our study indicates that controlling the phase structure of CS is a promising method to modulate the bioactivity of polymer microsphere system for potential bone tissue regeneration

Bioactivity of tempe by inhibiting adhesion of ETEC to intestinal cells, as influenced by fermentation substrates and starter pure cultures

Soya bean tempe is known for its bioactivity in reducing the severity of diarrhoea in piglets. This bioactivity is caused by an inhibition of the adhesion of enterotoxigenic Escherichia coli (ETEC) to intestinal cells. In this paper, we assessed the bioactive effect of soya tempe on a range of ETEC target strains, as well as the effect of a range of cereal and leguminous substrates and starter pure cultures. Soya bean tempe extracts strongly inhibited the adhesion of ETEC strains tested. All tempe made from other leguminous seeds were as bioactive as soya bean tempe, whereas tempe made from cereals showed no bioactivity. Using soya beans as substrate, fermentation with several fungi (Mucor, Rhizopus spp. and yeasts) as well as Bacillus spp. resulted in bioactive tempe, whereas fermentation with lactobacilli showed no bioactivity. The active component is releasedor formed during the fermentation and is not present in microbial biomass and only partly in unfermented substrates. The bioactivity being not specific for a single ETEC strain, makes the bioactive tempe relevant for applications in animal husbandry

A screen of Crohn's disease-associated microbial metabolites identifies ascorbate as a novel metabolic inhibitor of activated human T cells.

Microbial metabolites are an emerging class of mediators influencing CD4+ T-cell function. To advance the understanding of direct causal microbial factors contributing to Crohn's disease, we screened 139 predicted Crohn's disease-associated microbial metabolites for their bioactivity on human CD4+ T-cell functions induced by disease-associated T helper 17 (Th17) polarizing conditions. We observed 15 metabolites with CD4+ T-cell bioactivity, 3 previously reported, and 12 unprecedented. A deeper investigation of the microbe-derived metabolite, ascorbate, revealed its selective inhibition on activated human CD4+ effector T cells, including IL-17A-, IL-4-, and IFNγ-producing cells. Mechanistic assessment suggested the apoptosis of activated human CD4+ T cells associated with selective inhibition of energy metabolism. These findings suggest a substantial rate of relevant T-cell bioactivity among Crohn's disease-associated microbial metabolites, and evidence for novel modes of bioactivity, including targeting of T-cell energy metabolism

Preparation of bioactive surface via gel oxidation on titanium for biomedical application (hip joint replacement)

Titanium and its alloys are widely used as implant in biomedical applications. They have good mechanical and chemical properties, biocompatibility and biointegration with human body, but they have no ability to bond directly to natural bone. Therefore, alkali and heat treatments (gel oxidation) were introduced to improve the bioactivity of titanium by forming a mixture of sodium titanate and rutile on the surface of titanium. This method enables titanium to possess a bioactive surface which is essential to induce the apatite formation. This study aims to investigate the effects of alkali, sodium removal and heat treatments on in vitro bioactivity of titanium. UV light irradiation was used to study the effect on in vitro bioactivity of titanium. Alkalitreated titanium subjected to heat treatment in air have shown better overall in vitro performance than those treated in argon atmosphere. Therefore, the sodium removal treatment (dilute hydrochloric acid (HCl) treatment) was introduced to convert sodium titanate into anatase to improve the bioactivity of titanium treated in argon atmosphere. Thus, four samples (AT-0.5-HT500R, AT-0.5-HT600R, AT-5-HT500R and AT-5- HT600R) with different ratios of anatase to rutile were produced by varying the concentration of HCl acid treatment and heating temperature in argon atmosphere. It was found that the incorporation of sodium removal treatment has reduced two times the duration of apatite formation as compared with the conventional alkali and heat treatments. In order to further enhance the bioactivity, these samples were subjected to six different conditions of ultraviolet light irradiation and followed by in vitro bioactivity test. As a result, AT-5-HT500R (82.2% anatase and 17.8% rutile) was proven to deliver the best performance. It was confirmed that UV light irradiation enhances the bioactivity by removing hydrocarbon, inducing superhydrophilicity and forming OH groups. It was discovered that the duration of apatite formation was shortened to 7 days. Furthermore, the continuous UVA irradiation during in vitro test resulted in the acceleration of bonelike apatite formation in 3 days. It can be concluded that the sodium removal treatment and UV light irradiation give very significant impact to the formation of bonelike apatite on the titanium surfaces for biomedical applications

Research Article

published quarterly. The aim of IJPBS is to publish. peer reviewed research and review articles rapidly without delay in the developing field of pharmaceutical and biological science

Bioactivity of wollastonite/aerogels composites obtained from a TEOS-MTES matrix

Organic-inorganic hybrid materials were synthesized by controlled hydrolysis of tetraethoxysilane (TEOS), methyltrimethoxysilane (MTES), synthetic wollastonite powders and polydimethylsiloxane (PDMS) in an ethanol solution. Aerogels were prepared from acid hydrolysis of TEOS and MTES with different volume ratio in ethanol, followed by addition of wollastonite powder and PDMS in order to obtain aerogels with 20 wt% of PDMS and 5 wt% of CaO of the total silica. Finally, when the wet gels were obtained, they were supercritically dried at 260°C and 90 bar, in ethanol. In order to obtain its bioactivity, one method for surface activation is based on a wet chemical alkaline treatment. The particular interest of this study is that we introduce hybrid aerogels, in a 1 M solution of NaOH, for 30 s at room temperature. We evaluate the bioactivity of TEOS-MTES aerogel when immersed in a static volume of simulated body fluid (SBF). An apatite layer of spherical-shaped particles of uniform size smaller than 5 microns is observed to form on the surface of the aerogels after 25 days soaking in SBF.Ministerio de Ciencia e Innovación MAT2005-01583Junta de Andalucía TEP 79

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

Sex hormone-binding globulin regulation of androgen bioactivity in vivo : validation of the free hormone hypothesis

Sex hormone-binding globulin (SHBG) is the high-affinity binding protein for androgens and estrogens. According to the free hormone hypothesis, SHBG modulates the bioactivity of sex steroids by limiting their diffusion into target tissues. Still, the in vivo physiological role of circulating SHBG remains unclear, especially since mice and rats lack circulating SHBG post-natally. To test the free hormone hypothesis in vivo, we examined total and free sex steroid concentrations and bioactivity on target organs in mice expressing a human SHBG transgene. SHBG increased total androgen and estrogen concentrations via hypothalamic-pituitary feedback regulation and prolonged ligand half-life. Despite markedly raised total sex steroid concentrations, free testosterone was unaffected while sex steroid bioactivity on male and female reproductive organs was attenuated. This occurred via a liganddependent, genotype-independent mechanism according to in vitro seminal vesicle organ cultures. These results provide compelling support for the determination of free or bioavailable sex steroid concentrations in medicine, and clarify important comparative differences between translational mouse models and human endocrinology

Bioactivity, persistence and mobility of picloram in Selangor and Serdang soil series

The effects of environmental factors on bioactivity, persistence and mobility of picloram were studied in the laboratory and greenhouse using long beans (Vigna sinensis Endl. ex Hassk), as a bioassay species. Two soils were used, namely Selangor series (silty clay loam soil) and Serdang series (silty clay soil). The bioactivity of picloram (Tordon®) was inversely correlated with the organic matter content of the soil and increased with increasing herbicide concentrations. The half-life ofpicloram decreased from 33.1 to 24.1 days as soil temperature increased from 25°C to 35°C in the Selangor series, and from 28.1 to 17.7 days in the Serdang series soil. The degradation rate of picloram also increased with increasing moisture level in both soil series. However, picloram persisted longa in Selangor series soil than in Serdang series. Downward mobility through the soil profile was inversely related to the organic maller content of the soil. The quantity and frequency of simulated rain directly influenced the extent of leaching of the herbicide. Downward movement of picloram was greater in Serdang series which contains less organic matter but high sand than Selangor series longest available vessel