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.

A novel animal model of underactive bladder: analysis of lower urinary tract function in a rat lumbar canal stenosis model.

AIMS: An animal model of neurogenic underactive bladder (UAB) has not been established. It was reported that a rat lumbar spinal canal stenosis (LCS) model created by cauda equina compression manifested intermittent claudication and allodynia. In this study, we examined the lower urinary tract function of the rat LCS model. METHODS: One small hole was drilled at the fifth lumbar vertebral arch (sham), and a rectangular piece of silicone rubber was inserted into the L5-L6 epidural space (LCS). Before and after surgery, a metabolic cage study was performed. After surgery, awake cystometry (CMG) and an in vitro muscle strip study were performed. Bladder morphology was evaluated by hematoxylin and eosin staining. RESULTS: The LCS rats showed a significant decrease in voided volume and a significant increase in postvoid residual volume and residual urine rate compared with Sham rats. CMG showed that the postvoid residual urine volume and numbers of non-voiding contractions significantly increased, while the voided volume, threshold pressure, and maximum intravesical pressure during voiding significantly decreased. There were no significant differences between sham and LCS rats in response to carbachol. In contrast, there was a significant increase in response to field stimulation, especially at lower frequencies, in LCS rats. LCS rats showed no obvious difference in detrusor morphology. CONCLUSIONS: This rat model requires a relatively simple surgical procedure and has characteristics of neurogenic UAB. It seems to be useful in the pathophysiological elucidation of UAB and might have potential for assessment of pharmacotherapy of UAB.

Affinity Diversification of a Polymer Probe for Pattern-recognition-based Biosensing Using Chemical Additives.

Pattern-recognition-based sensing has attracted attention as a promising alternative to conventional sensing methods that rely on selective recognition. Here, we report on novel strategy using chemical additives with the ability to modulate probe/analyte interactions to more easily construct pattern-recognition-based sensing systems for proteins and cells. The fluorescence of dansyl-modified cationic poly-L-lysine (PLL-Dnc) is enhanced upon binding to proteins in aqueous solution, while the addition of salts, inert polymers, or alcohols modulates the protein/PLL-Dnc interactions via a variety of mechanisms. Subsequent readout of the fluorescence changes produces response patterns that reflect the characteristics of the analytes. Multivariate analysis of the response patterns allowed for accurate identification of not only eight structurally similar albumin homologues, but also four mammalian cells. This strategy, which uses inexpensive and common additives, significantly improves the accessibility of pattern-recognition-based sensing, which will offer new opportunities for the detection of various bioanalytes.

Rinse and evaporation coating of poly(methyl methacrylate) microchip for separation of sodium dodecyl sulfate-protein complex.

We developed a novel channel wall coating on a poly(methyl methacrylate) (PMMA) microchip using methylcellulose (MC) as a coating reagent to suppress electroosmotic flow (EOF) following the strong analytes adsorption via hydrophobic interaction with channel walls of PMMA. Our coating was obtained by first rinsing channel walls with MC-containing aqueous solution followed by evaporation. The coating made the hydrophilic channel wall lowering EOF by two orders of magnitude (1.2 x 10(-5)cm(2)V(-1)s(-1)) as well as reducing the hydrophobic adsorption. On the coated channel walls, we successfully separated sodium dodecyl sulfate-protein complexes with high reproducibility and efficiency using dextran as a lower viscosity protein separation medium.

Poly(methylmethacrylate) microchip electrophoresis of proteins using linear-poly(acrylamide) solutions as separation matrix.

Poly(methylmethacrylate) (PMMA) microchip electrophoresis of sodium dodecyl sulfate-protein complexes (SDS-PC) using linear-poly(acrylamide) (L-PA) as a separation matrix was investigated. Prior to electrophoresis, channel walls of PMMA were modified with methylcellulose (MC) to prevent adsorption between channel walls and SDS-PC. Size-based protein separation (SBPS) was successfully performed using the MC-coated microchips with Ferguson plot-fittings. The entangled L-PA solution provided high resolution of peaks of SDS-PC when the concentration of L-PA was increased. Some investigations into the separation mechanism, such as the plot of the logarithm of mobility of each SDS-PC versus the logarithm of the molecular weight of the complex exhibiting linear behavior, indicated that the separation mechanism was dependent on mass discrimination, in accordance with Ogston model.

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.

A risk factor analysis of accumulated postoperative pain and swelling sensation after dental implant surgery using a cellular phone-based real-time assessment.

PURPOSE: The purpose of this study was to identify the related risk factors of dental implant accumulated postoperative pain and swelling by cellular phone-based assessment. METHODS: Subjects were a consecutive series of patients who received oral implant surgery at Okayama University Hospital. Cellular phone-based questionnaire was sent at pre-set schedule to each subject every 2h on the day of surgery, and every 24h from the 2nd to 7th day post-surgery. Subjects replied in real-time the pain and swelling levels at the operated sites by an 11- and 4-grade rating-scale questionnaire. Overall intensity of individual pain and swelling was calculated by means of area under curve that drew by their time-dependent changes. Predictor variables were age, gender, presence of diabetes mellitus and/or hypertension (DM/HT), history of implant surgery, number of inserted implants, flap operation, surgical duration, pre-surgery anxiety, osteoplasty, bone quality, premedication, dosage of prescribed analgesics and local anesthesia and accumulated postoperative pain/swelling. Compliance rate and risk factors correlated with accumulated postoperative pain and swelling were calculated by multiple regression analysis. RESULTS: Final subjects were 18 females and 7 male (mean age: 59.3±7.32 yrs). Significant factors correlated with accumulated postoperative pain were DM/HT, surgical duration, premedication, bone quality, pre-surgery anxiety and postoperative swelling (R(2)=0.769, p=0.001, 0.013, 0.032, 0.007, 0.035 and 0.007, respectively). Meanwhile, significant factors associated with postoperative swelling were postoperative pain, DM/HT and bone quality (R(2)=0.365, p=0.002, 0.004, 0.008, respectively). CONCLUSION: These results suggested DM/HT and bone quality are correlated to overall intensity of postoperative pain and swelling.

[A case of pneumoconiosis in a dental technician].

We report a case of pneumoconiosis in a dental technician. He was a 33-year-old man who had worked in a dental clinic as a dental technician for 12 years. In October 1999, he visited to the National Zentsuji Hospital complaining of progressive cough and sputum over a three-year period. Although he received medication, his condition did not improve. He visited the same hospital again on May 8, 2000. His chest radiographs and CT films showed massive shadows in both upper lung lobes. Pneumoconiosis was diagnosed from the pathological findings in a lung specimen obtained by video-assisted thoracic surgery (VATS). There are few reports of pneumoconiosis of dental technicians diagnosed by VATS.

Power-free microchip immunoassay of PSA in human serum for point-of-care testing.

For point-of-care testing (POCT), a sandwich-typed immunoassay on poly(dimethylsiloxane) (PDMS) microfluidic chips was developed by employing two original technologies: a power-free microchip and laminar flow-assisted dendritic amplification (LFDA). Sequential injection of reagents of immunoassay was carried out by the power-free microchip. In addition, for the amplification of a fluorescent signal, LFDA exploited the interaction between FITC-labeled streptavidin and biotinylated anti-streptavidin on the interface of the two streams of laminar flow during the sequential injection of the reagents of the immunoassay. The microfluidic immunoassay was finished within 21 min with a sample volume of 500 nL. The calibration curve of a quantitative analysis satisfied the concentration range of the cut-off values of prostate-specific antigen (PSA) in serum. The limit of detection (LOD) reached 520 pg/mL (16 pM) of PSA in human female serum. We believe that this microfluidic immunoassay and device are promising for POCT.

Channel wall coating on a poly-(methyl methacrylate) CE microchip by thermal immobilization of a cellulose derivative for size-based protein separation.

We demonstrate channel wall coating using a cellulose derivative on a poly-(methyl methacrylate) (PMMA) CE microchip to eliminate EOF disturbing protein separation. The channel walls were modified by preconditioning with a solution containing the cellulose derivative and then thermally evaporating the solution to produce hydrophilic channel walls which prevent adsorption of analytes via a hydrophobic interaction. When the PMMA substrate was coated with the cellulose derivative hydroxypropylmethylcellulose (HPMC) 90SH, the water contact angle on the coated substrate was decreased (up to 15 degrees ) and EOF was significantly suppressed (up to 4.0 x 10(-6) cm2.V(-1)s(-1)). Three proteins (20.5, 68.0, and 114.6 kDa) were successfully separated on the 0.15% HPMC 90SH-coated channel walls with good reproducibility of migration time (RSD <1.75%) and high efficiency (theoretical plate number per meter: 2.62 x 10(5)).