Tunable Plasmonic Silver Nanodomes for Surface-Enhanced Raman Scattering

Surface-enhanced Raman scattering (SERS) is an emerging analytical method used in biological and non-biological structure characterization. Since the nanostructure plasmonic properties is a significant factor for SERS performance, nanostructure fabrication with tunable plasmonic properties are crucial in SERS studies. In this study, a novel method for fabrication of tunable plasmonic silver nanodomes (AgNDs) is presented. The convective-assembly method is preferred for the deposition of latex particles uniformly on a regular glass slide and used as a template for polydimethylsiloxane (PDMS) to prepare nanovoids on a PDMS surface. The obtained nanovoids on the PDMS are used as a mold for AgNDs fabrication. The nanovoids are filled with Ag deposition by the electrochemical method to obtain metallic AgNDs. Scanning electron microscopy (SEM) and atomic force microscopy (AFM) are used for characterization of the structural properties of all fabricated AgNDs. The optical properties of AgNDs are characterized with the evaluation of SERS activity of 4-aminothiphonel and rhodamine 6G. In addition to experimental characterizations, the finite difference time domain (FDTD) method is used for the theoretical plasmonic properties calculation of the AgNDs. The experimental and theoretical results show that the SERS performance of AgNDs is strongly dependent on the heights and diameters of the AgNDs.     

Tricine-SDS-PAGE

Tricine-SDS-PAGE is commonly used to separate proteins in the mass range 1-100 kDa. It is the preferred electrophoretic system for the resolution of proteins smaller than 30 kDa. The concentrations of acrylamide used in the gels are lower than in other electrophoretic systems. These lower concentrations facilitate electroblotting, which is particularly crucial for hydrophobic proteins. Tricine-SDS-PAGE is also used preferentially for doubled SDS-PAGE (dSDS-PAGE), a proteomic tool used to isolate extremely hydrophobic proteins for mass spectrometric identification, and it offers advantages for resolution of the second dimension after blue-native PAGE (BN-PAGE) and clear-native PAGE (CN-PAGE). Here I describe a protocol for Tricine-SDS-PAGE, which includes efficient methods for Coomassie blue or silver staining and electroblotting, thereby increasing the versatility of the approach. This protocol can be completed in 1-2 d.     

Tailed primer base excision sequence scanning (TP-BESS) for detection of single nucleotide polymorphisms (SNPs)

Base excision sequence scanning (BESS) is used to detect and localise point mutations in mammalian genes. The cost of BESS can be significant in large-scale projects. however, due to the requirement for end-labelling of one of the two PCR primers. This is especially true when a fluorescent label is used for detection. If a universal label could be used for all of the PCR reactions, it would reduce the cost of this technique significantly. Here we describe a TP-BESS procedure where one fluorescence-labelled primer is used as a universal label for all the PCR reactions in BESS analysis. The universal label makes BESS financially more feasible for large-scale detection of single nucleotide polymorphisms (SNPs).     

Surveillance in longitudinal models: detection of intrauterine growth restriction

A new methodology for online detection of intrauterine growth restriction (IUGR) is proposed where traditional methods for statistical surveillance are applied. Here, deficient growth rate is used to detect IUGR instead of the common surrogate measure "small for gestational age" (SGA). Fetal growth is estimated by repeated measurements of symphysis-fundus (SF) height. At each time point the new method, based on the Shiryaev-Roberts method, is used to evaluate the growth in SF height. We use Swedish data to model a normal growth pattern, which is used to evaluate the capability of the new method to detect IUGR in comparison with a method used in practice today. Results from simulations indicate that the new method performs considerably better than the method used today. We also illustrate the effect of some important factors which influence the detection ability and illuminate the tendency of the method used today to misclassify SGA cases as IUGR.     

A novel microfluidic biosensor based on an electrical detection system for alpha-fetoprotein

Conventional immunoassays are labor intensive, expensive and time consuming and require large pieces of equipment for detection. Therefore, we have developed and characterized a novel immunoassay methodology comprised of microbeads and microbiochips. In this method, microbeads are used to filter and immobilize antibodies and an immuno-gold silver staining (IGSS) method is then used to amplify electrical signals that correspond to the bound antibodies. The chip used for this system is composed of an inexpensive and biocompatible polydimethylsiloxane (PDMS) layer over a Pyrex glass substrate that contains a platinum (Pt) microelectrode, which is used to detect the electrical signal in this system, the microelectrode is fabricated on the substrate and a microchannel and pillar-type microfilter is formed in the PDMS layer. A sandwich immunoassay approach was applied to detect alpha-fetoprotein (AFP), a cancer biomarker, using this system. The results of this study showed that the time required for a complete assay was reduced by 1h and a detection limit as low as 1 ng/mL was attained when this system used, which indicates that similar bead-based electrical detection systems could be used for the diagnosis of many forms of cancer.     

Separation and quantitation ofcis- andtrans-thiothixene in human plasma by high-performance liquid chromatography

A high-performance liquid chromatographic procedure is described for the separation ofcis andtrans isomers of thiothixene, a thioxanthene derivative used as an antipsychotic agent. A radial compression module (RCM-100) was used with both silica and cyanopropyl cartridges. A fixed-wavelength UV detector (254 nm) was used in these studies for quantitation. Mesoridazine is used as an internal standard because of its separation characteristics and reproducible quantitation. C₁₈ Sep-Pak cartridges are used for biological sample cleanup. Plasma samples from patients treated with thiothixene (Navane) were assayed forcis andtrans-thiothixene. Notrans-thiothixene was detectable andcis-thiothixene concentrations ranged from 0 to 22.5 ng/ml.     

Rapid simulation of P values for product methods and multiple-testing adjustment in association studies

A major aim of association studies is the identification of polymorphisms (usually SNPs) associated with a trait. Tests of association may be based on individual SNPs or on sets of neighboring SNPs, by use of (for example) a product P value method or Hotelling's T test. Linkage disequilibrium, the nonindependence of SNPs in physical proximity, causes problems for all these tests. First, multiple-testing correction for individual-SNP tests or for multilocus tests either leads to conservative P values (if Bonferroni correction is used) or is computationally expensive (if permutation is used). Second, calculation of product P values usually requires permutation. Here, we present the direct simulation approach (DSA), a method that accurately approximates P values obtained by permutation but is much faster. It may be used whenever tests are based on score statistics--for example, with Armitage's trend test or its multivariate analogue. The DSA can be used with binary, continuous, or count traits and allows adjustment for covariates. We demonstrate the accuracy of the DSA on real and simulated data and illustrate how it might be used in the analysis of a whole-genome association study.     

Basic studies and applications on bioremediation of DDT: A review

The persistent insecticide DDT (1,1,1-trichloro-2,2-bis (4-chlorophenyl) ethane) has been widely used for pest control in the management of mosquito-borne malaria and is still used for that purpose in some tropical countries. Considering the potential for negative effects due to DDT contamination, it is necessary to determine effective methods of remediation. Several methods have been used to degrade or transform DDT into less toxic compounds. Bacteria and white-rot fungi (WRF) have been shown to enhance the degradation process in soil using both pure and mixed cultures. Recently, a biological approach has been used as an environmentally-friendly treatment, using new biological sources to degrade DDT, e.g. brown-rot fungi (BRF), cattle manure compost (CMC) and spent mushroom waste (SMW). In this review, the abilities of BRF, CMC and SMW to degrade DDT are discussed, including the mechanisms and degradation pathways. Furthermore, application of these sources to contaminated soil is also described. The review discusses which is the best source for bioremediation of DDT.     

Characterization of the microbial community in indoor environments: a chemical-analytical approach

An integrated procedure is presented whereby gas chromatography-ion trap mass spectrometry is used to determine chemical markers of gram-negative bacterial lipopolysaccharide (3-hydroxy fatty acids with 10 to 18 carbon atoms), gram-positive bacteria (branched-chain fatty acids with 15 and 17 carbon atoms), bacterial peptidoglycan (muramic acid), and fungal biomass (ergosterol) in samples of settled house dust. A hydrolysate of (13)C-labeled cyanobacterial cells is used as an internal standard for the first three markers. These analyses require two dust samples, one for 3-OH fatty acids, branched-chain fatty acids, and muramic acid and another for ergosterol. The method may be used to characterize microbial communities in environmental samples.     

High performance thin layer chromatography (HPTLC), an improved technique for screening lichen substances

Abstract: High performance thin layer chromatography (HPTLC) is a method that can be used for screening lichen substances. It is as simple to use as standard TLC, but has many advantages: It is more sensitive, it is possible to run more samples in a shorter period of time, and the amount of solvent used is much smaller. The material needed and the methods used are described in detail. Horizontal chromatogram development was used. Since two of the solvents used in system B have been substituted, and since the properties of the HPTLC plates are slightly different, our results are not entirely in accordance with the standardized TLC method. A revised table for the identification of 69 lichen substances (obtained from 62 taxa) is accordingly presented.     

A neural network model for reconstructing EMG signals from eight shoulder muscles: consequences for rehabilitation robotics and biofeedback

This paper demonstrates the ability of a fully connected feed forward neural network (FFNN) using the backpropagation training algorithm to predict the electromyography (EMG) signal from eight muscles of the shoulder for both exercises not used for training and EMG signals from subjects not used for training. The network showed a good predictive ability for subjects not used for training (r(2) between 0.33 and 0.84) and for activities not used for training (r(2) between 0.56 and 0.89). This may have applications for patients, physical therapists and doctors to monitor patient performance by reviewing the level of agreement between the patient EMG and the predicted EMG. Coupled with traditional methods of evaluation, EMG can provide an excellent measure of how well a patient has responded or is responding to treatment. Incorporating robotic technology could facilitate the use of EMG as an input to an intelligent decision making algorithm used to increase or decrease the level of difficulty according to patient performance.     

SOI-nanowire biosensor for detection of D-NFATc1 protein

The nanowire (NW) detection is one of the fast-acting and high-sensitive methods, which can recognize potentially relevant protein molecules. A NW-biosensor based on the silicon-on-insulator (SOI)-structures has been used for biospecific label-free real time detection of the NFATc1 (D-NFATc1) oncomarker. For this purpose, SOI-nanowires (NWs) were modified with aptamers against NFATc1 used as molecular probes. It was shown that using this biosensor it is possible to reach sensitivity of 10^?15 M. This sensitivity was comparable to that of the NW-biosensor with immobilized antibodies used as macromolecular probes. The results demonstrate that approaches used in this study are promising for development of sensor elements for high-sensitive diagnostics of diseases.     

Chemiluminescent and bioluminescent assays as innovative prospects for mycotoxin determination in food and feed.

Mycotoxin contamination of food and feedstuffs is among the top priorities for human and animal safety. The currently used techniques for mycotoxin determination, either chromatography or ELISA, are unsuitable for routine in-field assessment. There is an urgent need for other accurate, simple and cost-effective techniques that can be used as a screening tool for a rapid estimation of mycotoxin contamination in commodity lots. This paper reviews the literature on the use of chemiluminescence (CL) and bioluminescence (BL) assays for direct or indirect mycotoxin assessment. The chemiluminescence immunoassays, adenosine triphosphate (ATP) bioluminescence and bioassays are reviewed and their advantages and limitations discussed. These techniques used in food testing and the pharmaceutical industry offer promise as rapid techniques for mycotoxin determination. Chemiluminescence and bioluminescence bioassays are the most innovative alternatives to the conventional techniques used for mycotoxin determination in food and feed.     

Cometabolic degradation of trichloroethylene in a bubble column bioscrubber

A bubble column bioreactor was used as bioscrubber to carry out a feasibility study for the cometabolic degradation of trichloroethylene (TCE). Phenol was used as cosubstrate and inducer. The bioreactor was operated like a conventional chemostat with regard to the cosubstrate and low dilution rates were used to minimize the liquid outflow. TCE degradation measurements were carried out using superficial gas velocities between 0.47and 4.07 cm s(-1) and TCE gas phase loads between 0.07 and 0.40 mg L(-1) Depending on the superficial gas velocity used, degrees of conversion between 30% and 80% were obtained. A simplified reactor model using plug flow for the gas phase, mixed flow for the liquid phase, and pseudo first order reaction kinetics for the conversionof TCE was established. The model is able to give a reasonable approximation of the experimental data. TCE degradation at the used experimental conditions is mainly limited by reaction rate rather than by mass transfer rate. The model can be used to calculate the reactor volume and the biomass concentration for a required conversion. (c) 1995 John Wiley & Sons Inc.     

The effect of primary fixation with standard postfixation and the duration of hydrolysis on nuclear features in image cytometry.

The effect of three primary fixation procedures, used in the preparation of routine cytological samples: air-drying, Delaunay, and Saccomanno fixation, with postfixation in modified B?hm-Sprenger fixative, on nuclear features as a function of hydrolysis time is reported. Three different cell types: lymphatic cells (tonsil), epithelial cells (buccal mucosa) and mesenchymal cells (uterine myometrium) were used for the study. Our findings show, that generally not all features have the same plateau times as the IOD (integrated optical density), and that many features show different values depending on cell type and fixation method. It is therefore recommended that for any primary fixative used in routine clinical work and for each cell type, the hydrolysis curve for all nuclear features to be used in sample analysis should be established.     

Chitosan Interactions with Metal Ions and Dyes: Dissolved-state vs. Solid-state Application

Summary Chitosan is an amino-polysaccharide with highly efficient properties for the binding of metal ions and anionic dyes. Uptake may occur through chelation on free amino functions (at near-neutral pH) or by electrostatic attraction on protonated amino groups (in acidic solutions). The polymer is soluble in acidic solutions and its binding properties can be used in both solid form (sorption) and liquid form (ultrafiltration coupled with chelation, coagulation–flocculation). These properties have been used for the recovery of mercury from dilute solutions at initial pH 5 (which reveals the most efficient pH in the range pH 4–6) and for the recovery of Reactive Black 5 (RB5, anionic dye) at pH 3. While in the case of mercury binding saturation of the biopolymer is only slightly higher when chitosan is used in the liquid form compared to solid-state adsorption, in the case of the coagulation–flocculation of RB5 (using the liquid-form of chitosan) the saturation of the polymer (calculated on the basis of molar ratio of dye vs. amino groups of the polymer) is reached at a significantly greater value than when the polymer is used for the solid-state binding of the dye. There is a much more efficient use of amino groups when chitosan is used in the liquid-form due to a better availability of amino groups (less hydrogen bonds between the chains of the polymer) and to a better accessibility to internal sorption sites (lower diffusion control).     

Use of Kampo Diagnosis in Randomized Controlled Trials of Kampo Products in Japan: A Systematic Review

Background

The Committee for Evidence-based Medicine (EBM) of the Japan Society for Oriental Medicine started compiling Evidence Reports of Kampo Treatment (EKAT) in 2007. EKAT is a compilation of structured abstracts of randomized controlled trials (RCTs), along with comments by a third party reviewer. As of 31 December, 2012, there were 378 RCTs of Kampo medicines in Japan. The primary research question of this study is “How frequently is Kampo diagnosis used in RCTs of Kampo medicines?” The secondary research question is “When is Kampo diagnosis used in RCTs?”

Materials and Methods

The structured abstract (SA) of each RCT article was reviewed to examine how Kampo diagnosis was used in RCTs, especially how Kampo diagnosis was used in the randomization process.

Results

Kampo diagnosis was used before randomization in 27 RCTs (7.1%), after randomization in 31 RCTs (8.2%), and not used in 320 RCTs (84.7%). Before randomization, Kampo diagnosis was used as a criterion for inclusion in 10 RCTs, criterion for exclusion in 9 RCTs, and criteria for both inclusion and exclusion in 2 RCTs. Kampo formulas were determined according to Kampo diagnosis in 7 RCTs. After randomization, subgroup analyses according to Kampo diagnosis were done in 27 RCTs, and grade of disease severity at Kampo diagnosis was used for analysis as an endpoint in 4 RCTs.

Conclusions

Kampo diagnosis was used before randomization only in approximately 15% of RCTs, and the number of RCT articles using Kampo diagnosis after randomization was almost the same as that before randomization. Further studies to determine the good RCTs conforming to CONSORT requirements and good systematic reviews conforming to PRISMA requirements are needed to clarify the significance of Kampo diagnosis.     

Discrimination of Parkinsonian tremor from essential tremor by implementation of a wavelet-based soft-decision technique on EMG and accelerometer signals

A wavelet-decomposition with soft-decision algorithm is used to estimate an approximate power-spectral density (PSD) of both accelerometer and surface EMG signals for the purpose of discrimination of Parkinson tremor from essential tremor. A soft-decision wavelet-based PSD estimation is used with 256 bands for a signal sampled at 800 Hz. The sum of the entropy of the PSD in band 6 (7.8125–9.375 Hz) and band 11 (15.625–17.1875 Hz) is used as a classification factor. The data has been recorded for diagnostic purposes in the Department of Neurology of the University of Kiel, Germany. Two sets of data are used. The training set, which consists of 21 essential-tremor (ET) subjects and 19 Parkinson-disease (PD) subjects, is used to obtain the threshold value of the classification factor differentiating between the two subjects. The test data set, which consists of 20 ET and 20 PD subjects, is used to test the technique and evaluate its performance. A “voting” between three results obtained from accelerometer signal and two EMG signals is applied to obtain the final discrimination. A total accuracy of discrimination of 85% is obtained.     

巴戟天应用的名实沿革考

通过对古今文献的分析及实地考查认为:南北朝以前使用之主流巴戟天药材的原植物可能为五味子科植物铁箍散Schisandr a propinqua(Wall.)Baill.var.sinensis Oliv.,药材现称川巴戟或香巴戟;唐代至清末广为运用的品种为归州巴戟天,原植物为茜草科植物四川虎刺Dammacanthus officinarum Huang,药材现称鄂西巴戟天或恩施巴戟天;现今药典收载之巴戟天为清末发展的新品种,其原植物为茜草科植物巴戟天 Morinda officinalis How。     

Two inhibitors of DNA-synthesis lead to inhibition of cytokine production via a different mechanism

Methotrexate (MTX) and mycophenolic acid (MPA) are used in the clinic for their immunosuppressive properties. MTX is widely used for the treatment of rheumatoid arthritis (RA). MPA is used to prevent graft rejection and is now experimentally used in systemic lupus erythematosis and RA. It is known that both drugs interfere with DNA synthesis. However, the precise mechanism of action is still debated. We have analysed the effect of the drugs on cytokine production in whole blood during short cultures. The production of T-cell cytokines was inhibited by both drugs. MTX inhibits cytokine production because MTX induces apoptosis in activated T-cells. MPA inhibits cytokine production by preventing T-cells to progress to the S-phase of the cell cycle. Cytokine production by monocytes was slightly decreased by the drugs. The reason for this inhibition is not clear. These results indicate that T-cells are the main target cells of the immunosuppressive drugs MPA and MTX.