Functional annotation of chemical libraries across diverse biological processes.

Chemical-genetic approaches offer the potential for unbiased functional annotation of chemical libraries. Mutations can alter the response of cells in the presence of a compound, revealing chemical-genetic interactions that can elucidate a compound's mode of action. We developed a highly parallel, unbiased yeast chemical-genetic screening system involving three key components. First, in a drug-sensitive genetic background, we constructed an optimized diagnostic mutant collection that is predictive for all major yeast biological processes. Second, we implemented a multiplexed (768-plex) barcode-sequencing protocol, enabling the assembly of thousands of chemical-genetic profiles. Finally, based on comparison of the chemical-genetic profiles with a compendium of genome-wide genetic interaction profiles, we predicted compound functionality. Applying this high-throughput approach, we screened seven different compound libraries and annotated their functional diversity. We further validated biological process predictions, prioritized a diverse set of compounds, and identified compounds that appear to have dual modes of action.

Can microcatheters be cleaned for reuse after NBCA embolization? Cleaning technique with gelatin sponge particles.

PURPOSE: To study a new technique for cleaning microcatheters for reuse after NBCA embolization ("NBCA"), and to evaluate the clinical reusability of microcatheters that were cleaned with gelatin sponge particles after NBCA. MATERIALS AND METHODS: Four cleaning solution flushes for microcatheters after NBCA injection-5 % glucose ("glucose") only, Lipiodol-glucose, gelatin sponge particles ("gelatin")-glucose, and Lipiodol-gelatin-glucose-were examined experimentally. These solutions were evaluated by performing three examinations: a microcatheter resistance test based on the time taken to pass water through the microcatheter, a microcatheter resistance test based on the ease of insertion of a microguidewire, and observations of the inner surfaces of the cylinders after NBCA. Microcatheters that had already been used in NBCA were cleaned using this new technique and then applied in 20 clinical sessions (19 patients). RESULTS: There was no significant difference in water passage time between the controls and the groups that received a gelatin flush. In the resistance test based on the insertion of a microguidewire, groups that received a gelatin flush showed significantly less resistance than the groups that did not receive a gelatin flush. Observations of the inner surfaces of the cylinders indicated that cleaning with gelatin can lead to inner surfaces that are almost indistinguishable from control surfaces in terms of cleanliness. All clinical procedures involving Lipiodol-gelatin-glucose flushes were performed without any technical difficulties or complications. CONCLUSIONS: Applying the new cleaning technique utilizing gelatin sponge particles to microcatheters after NBCA ensures that they are clean enough to be reused.

Plant-derived antifungal agent poacic acid targets ß-1,3-glucan.

A rise in resistance to current antifungals necessitates strategies to identify alternative sources of effective fungicides. We report the discovery of poacic acid, a potent antifungal compound found in lignocellulosic hydrolysates of grasses. Chemical genomics using Saccharomyces cerevisiae showed that loss of cell wall synthesis and maintenance genes conferred increased sensitivity to poacic acid. Morphological analysis revealed that cells treated with poacic acid behaved similarly to cells treated with other cell wall-targeting drugs and mutants with deletions in genes involved in processes related to cell wall biogenesis. Poacic acid causes rapid cell lysis and is synergistic with caspofungin and fluconazole. The cellular target was identified; poacic acid localized to the cell wall and inhibited ß-1,3-glucan synthesis in vivo and in vitro, apparently by directly binding ß-1,3-glucan. Through its activity on the glucan layer, poacic acid inhibits growth of the fungi Sclerotinia sclerotiorum and Alternaria solani as well as the oomycete Phytophthora sojae. A single application of poacic acid to leaves infected with the broad-range fungal pathogen S. sclerotiorum substantially reduced lesion development. The discovery of poacic acid as a natural antifungal agent targeting ß-1,3-glucan highlights the potential side use of products generated in the processing of renewable biomass toward biofuels as a source of valuable bioactive compounds and further clarifies the nature and mechanism of fermentation inhibitors found in lignocellulosic hydrolysates.

Image-based prediction of drug target in yeast.

Discovering the intracellular target of drugs is a fundamental challenge in biomedical research. We developed an image-based technique with which we were able to identify intracellular target of the compounds in the yeast Saccharomyces cerevisiae. Here, we describe the rationale of the technique, staining of yeast cells, image acquisition, data processing, and statistical analysis required for prediction of drug targets.

Factors affecting the use of complementary and alternative medicine among Japanese university students.

BACKGROUND: Patients suffering from intractable diseases and individuals seeking relief from mild symptoms resort to treatments outside the modern medical paradigm, such as complementary and alternative medicine (CAM). In order to improve doctor-patient communication about CAM, it is essential to evaluate CAM usage among social groups likely to choose it in the future. Therefore, we aimed to evaluate how university students - individuals highly subject to future CAM usage - perceive CAM and the factors affecting their choice of CAM use. METHODS: We conducted a questionnaire survey with 1,096 Japanese university students not studying medical subjects. RESULTS: The term CAM was known to 11% of the subjects. Modalities they most associated with CAM were art therapy (353 subjects), hot spring therapy (349), and aromatherapy (345). They had experience taking vitamins, trace elements, other supplements (498), and nutritional drinks (483). Several subjects wanted to experience shiatsu massage (373) and hot spring therapy (303). Multiple regression analysis of the modalities that the subjects wanted to experience revealed a 42% multiple coefficient of determination for prioritizing modalities that the subject associated with CAM, showing a large contribution of this deciding factor. CONCLUSIONS: Although most subjects were not familiar with the term CAM, many of them had decided to ingest substances in the CAM category on the basis of self-judgment and without adequate knowledge. Because such behavior can be detrimental to health, medical professionals should be aware of CAM usage among their patients and seek effective communication with them in order to enable safe CAM practice.