Metabolomics-guided isolation of anti-trypanosomal metabolites from the endophytic fungus Lasiodiplodia theobromae

Fungal endophytes offer diverse and unique secondary metabolites, making these organisms potential sources of promising drug leads. The application of high-resolution-liquid chromatography mass spectrometry and nuclear magnetic resonance-based metabolomics to fungal endophytes is practical in terms of dereplication studies and the mining of bioactive compounds. In this paper, we report the application of metabolomics in parallel with anti-trypanosomal assays to determine the ideal conditions for the medium-scale fermentation of the endophyte Lasiodiplodia theobromae. The (1)H NMR comparison between the active versus inactive fractions identified several unique chemical fingerprints belonging to the active fractions. Furthermore, by integrating high-resolution-liquid chromatography mass spectrometry data with multivariate data analysis, such as orthogonal partial least squares-discriminant analysis (OPLS-DA) and the bioactivity results of the fractions of L. theobromae, the anti-trypanosomal agents were easily discerned. With available databases such as Antibase and Dictionary of Natural Products coupled to MZmine through in-house algorithms optimized in our laboratory, the predicted metabolites were readily identified prior to isolation. Fractionation was performed on the active fractions and three known compounds were isolated, namely, cladospirone B, desmethyl-lasiodiplodin, and R-(-)-mellein. Cladospirone B and desmethyl-lasiodiplodin were among the predicted compounds generated by the OPLS-DA S-plot, and these compounds exhibited good activity against Trypanosoma brucei brucei with minimum inhibitory concentrations of 17.8 µM and 22.5 µM, respectively

Telehealth Services to Improve Nonadherence: A Placebo-Controlled Study

This is the publisher's version, also available electronically from http://online.liebertpub.com/doi/abs/10.1089/tmj.2006.12.289.The objective of this study was to test whether a telehealth intervention could improve the compliance with continuous positive airway pressure (CPAP) by patients with sleep apnea. These patients had been nonadherent for the initial 3 months of therapy even after receiving the initial standard and then supplemental audiotaped/videotaped patient education for adhering to CPAP nightly. The materials and methods included a randomized testing of experimental and placebo interventions. Interventions were delivered by nurses to two groups in their homes by telehealth over a 12-week period. The placebo intervention was used to control for Hawthorne effect, time and attention influences and the novelty of having telehealth in the home. Results following the telehealth interventions were that significantly more patients in the experimental group 1 (n = 10) than the placebo group 2 (n = 9) were adhering nightly to CPAP (χ2 = 4.55, p = 0.033). Group 1 patients reported greater satisfaction with their intervention. However, both groups rated telehealth delivery positively. The mean cost of each 20-minute telehealth visit was $30 while the total cost of the telehealth intervention for each patient was$420. These costs included telehealth equipment, initial installation, longdistance telephone charges, nurse salary, and intervention materials. Conclusions are that telehealth interventions are a potentially cost-effective service for increasing adherence to prescribed medical treatments. Replication studies with large samples and in other clinical groups are recommended

A comparison of walk-in counselling and the wait list model for delivering counselling services

Background: Walk-in counselling has been used to reduce wait times but there are few controlled studies to compare outcomes between walk-in and the traditional model of service delivery. Aims: To compare change in psychological distress by clients receiving services from two models of service delivery, a walk-in counselling model and a traditional counselling model involving a wait list Method: Mixed methods sequential explanatory design including quantitative comparison of groups with one pre-test and two follow ups, and qualitative analysis of interviews with a subsample. 524 participants 16 years and older were recruited from two Family Counselling Agencies; the General Health Questionnaire assessed change in psychological distress; prior use of other mental health and instrumental services was also reported. Results: Hierarchical linear modelling revealed clients of the walk-in model improved faster and were less distressed at the 4-week follow-up compared to the traditional service delivery model. At the 10-week follow-up, both groups had improved and were similar. Participants receiving instrumental services prior to baseline improved more slowly. Qualitative interviews confirmed participants valued the accessibility of the walk-in model. Conclusions: This study improves methodologically on previous studies of walk-in counselling, an approach to service delivery that is not conducive to randomized controlled trials

Metabolomic profiling of the synergistic effects of melittin in combination with cisplatin on ovarian cancer cells

Melittin, the main peptide present in bee venom, has been proposed as having potential for anticancer therapy; the addition of melittin to cisplatin, a first line treatment for ovarian cancer, may increase the therapeutic response in cancer treatment via synergy, resulting in improved tolerability, reduced relapse, and decreased drug resistance. Thus, this study was designed to compare the metabolomic effects of melittin in combination with cisplatin in cisplatin-sensitive (A2780) and resistant (A2780CR) ovarian cancer cells. Liquid chromatography (LC) coupled with mass spectrometry (MS) was applied to identify metabolic changes in A2780 (combination treatment 5 μg/mL melittin + 2 μg mL cisplatin) and A2780CR (combination treatment 2 μg/mL melittin + 10 μg/mL cisplatin) cells. Principal components analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) multivariate data analysis models were produced using SIMCA-P software. All models displayed good separation between experimental groups and high-quality goodness of fit (R2) and goodness of prediction (Q2), respectively. The combination treatment induced significant changes in both cell lines involving reduction in the levels of metabolites in the tricarboxylic acid (TCA) cycle, oxidative phosphorylation, purine and pyrimidine metabolism, and the arginine/proline pathway. The combination of melittin with cisplatin that targets these pathways had a synergistic effect. The melittin-cisplatin combination had a stronger effect on the A2780 cell line in comparison with the A2780CR cell line. The metabolic effects of melittin and cisplatin in combination were very different from those of each agent alone

Effect of bee venom and its fractions on the release of pro-inflammatory cytokines in PMA-differentiated U937 cells co-stimulated with LPS

The venom of Apis mellifera (honey bee) has been reported to play a role in immunotherapy, but existing evidence to support its immuno-modulatory claims is insufficient. Four fractions from whole bee venom (BV) were separated using medium pressure liquid chromatography. Their ability to induce the production of cytokines TNFα, IL-1β and IL-6 in phorbol-12-myristate-13-acetate (PMA)-treated U937 cells was assessed. The levels of the three cytokines produced by stimulation with the four fractions and crude BV without LPS were not significantly different from negative control values. However, co-stimulation of the cells with LPS and Fraction 4 (F-4) induced a 1.6-fold increase in TNF-α level (p < 0.05) compared to LPS alone. Likewise, LPS-induced IL-1β production was significantly synergised in the presence of F-1 (nine-fold), F-2 (six-fold), F-3 (four-fold) and F-4 (two-fold) fractions, but was only slightly enhanced with crude BV (1.5-fold) relative to LPS. Furthermore, the LPS-stimulated production of IL-6 was not significantly increased in cells co-treated with F-2 and F-3, but the organic fraction (F-4) showed an inhibitory effect (p < 0.05) on IL-6 production. The latter was elucidated by NMR spectroscopy and found to contain(Z)-9-eicosen-1-ol. The effects observed with the purified BV fractions were more marked than those obtained with the crude sample

MAMBO 1.2mm observations of luminous starbursts at z~2 in the SWIRE fields

We report on--off pointed MAMBO observations at 1.2 mm of 61 Spitzer-selected star-forming galaxies from the SWIRE survey. The sources are selected on the basis of bright 24um fluxes (f_24um>0.4mJy) and of stellar dominated near-infrared spectral energy distributions in order to favor z~2 starburst galaxies. The average 1.2mm flux for the whole sample is 1.5+/-0.2 mJy. Our analysis focuses on 29 sources in the Lockman Hole field where the average 1.2mm flux (1.9+/-0.3 mJy) is higher than in other fields (1.1+/-0.2 mJy). The analysis of the sources multi-wavelength spectral energy distributions indicates that they are starburst galaxies with far-infrared luminosities ~10^12-10^13.3 Lsun, and stellar masses of ~0.2-6 x10^11 M_sun. Compared to sub-millimeter selected galaxies (SMGs), the SWIRE-MAMBO sources are among those with the largest 24um/millimeter flux ratios. The origin of such large ratios is investigated by comparing the average mid-infrared spectra and the stacked far-infrared spectral energy distributions of the SWIRE-MAMBO sources and of SMGs. The mid-infrared spectra exhibit strong PAH features, and a warm dust continuum. The warm dust continuum contributes to ~34% of the mid-infrared emission, and is likely associated with an AGN component. This constribution is consistent with what is found in SMGs. The large 24um/1.2mm flux ratios are thus not due to AGN emission, but rather to enhanced PAH emission compared to SMGs. The analysis of the stacked far-infrared fluxes yields warmer dust temperatures than typically observed in SMGs. Our selection favors warm ultra-luminous infrared sources at high-z, a class of objects that is rarely found in SMG samples. Our sample is the largest Spitzer-selected sample detected at millimeter wavelengths currently available.Comment: Accepted for publication in ApJ (51 pages; 16 figures). The quality of some figures has been degraded for arXiv purposes. Full resolution version available at this http://www.iasf-milano.inaf.it/~polletta/mambo_swire/lonsdale08_ApJ_accepted.pd

Models for Multiband IR Surveys

Empirical 'backward' galaxy evolution models for IR-bright galaxies are constrained using multiband IR surveys. A new Monte-Carlo algorithm is developed for this task. It exploits a large library of realistic Spectral Energy Distributions (SEDs) of 837 local IR galaxies (IRAS 25$\mu m$ selected) from the UV (1000{\AA}) to the radio (20cm), including ISO-measured 3--13$\mu m$ unidentified broad features (UIBs). The basic assumption is that the local correlation between SEDs and Mid-Infrared (MIR) luminosities can be applied to earlier epochs of the Universe. Three populations of IR sources are considered in the evolution models. These include (1) starburst galaxies, (2) normal late-type galaxies, and (3) galaxies with AGN. A set of models so constructed are compared with data from the literature. Predictions for number counts, confusion limits, redshift distributions, and color-color diagrams are made for multiband surveys using the upcoming SIRTF satellite.Comment: 40 pages latex. 32 GIF figures. New Version (July 8, 2001) to be accepted by ApJ. High quality figures (included in a PS file of the paper) can be found in http://spider.ipac.caltech.edu/staff/cxu/papers/paper_model_3.ps.g

Communications Biophysics

Contains reports on eight research projects split into four sections.National Institutes of Health (Grant 5 P01 NS13126)National Institutes of Health (Grant 5 K04 NS00113)National Institutes of Health (Training Grant 5 T32 NS07047)National Science Foundation (Grant BNS80-06369)National Institutes of Health (Grant 5 ROl NS11153)National Institutes of Health (Fellowship 1 F32 NS06544)National Science Foundation (Grant BNS77-16861)National Institutes of Health (Grant 5 R01 NS10916)National Institutes of Health (Grant 5 RO1 NS12846)National Science Foundation (Grant BNS77-21751)National Institutes of Health (Grant 1 R01 NS14092)National Institutes of Health (Grant 2 R01 NS11680)National Institutes of Health (Grant 5 ROl1 NS11080)National Institutes of Health (Training Grant 5 T32 GM07301

Communications Biophysics

Contains reports on ten research projects.National Institutes of Health (Grant 5 P01 NS13126)National Institutes of Health (Training Grant 5 T32 NS0704)National Science Foundation (Grant BNS80-06369)National Institutes of Health (Grant 5 R01 NS11153)National Science Foundation (Grant BNS77-16861)National Institutes of Health (Grant 5 RO1 NS12846)National Science Foundation (Grant BNS77-21751)National Institutes of Health (Grant 1 P01 NS14092)Karmazin Foundation through the Council for the Arts at MITNational Institutes of Health (Fellowship 5 F32 NS06386)National Science Foundation (Fellowship SP179-14913)National Institutes of Health (Grant 5 RO1 NS11080

Chemical and antimicrobial profiling of propolis from different regions within Libya.

Extracts from twelve samples of propolis collected from different regions of Libya were tested for their activity against Trypanosoma brucei, Leishmania donovani, Plasmodium falciparum, Crithidia fasciculata and Mycobacterium marinum and the cytotoxicity of the extracts was tested against mammalian cells. All the extracts were active to some degree against all of the protozoa and the mycobacterium, exhibiting a range of EC50 values between 1.65 and 53.6 μg/ml. The toxicity against mammalian cell lines was only moderate; the most active extract against the protozoan species, P2, displayed an IC50 value of 53.2 μg/ml. The extracts were profiled by using liquid chromatography coupled to high resolution mass spectrometry. The data sets were extracted using m/z Mine and the accurate masses of the features extracted were searched against the Dictionary of Natural Products (DNP). A principal component analysis (PCA) model was constructed which, in combination with hierarchical cluster analysis (HCA), divided the samples into five groups. The outlying groups had different sets of dominant compounds in the extracts, which could be characterised by their elemental composition. Orthogonal partial least squares (OPLS) analysis was used to link the activity of each extract against the different micro-organisms to particular components in the extracts