Lateral flow devices for nucleic acid analysis exploiting quantum dots as reporters

There is a growing interest in the development of biosensors in the form of simple lateral flow devices that enable visual detection of nucleic acid sequences while eliminating several steps required for pipetting, incubation and washing out the excess of reactants. In this work, we present the first dipstick-type nucleic acid biosensors based on quantum dots (QDs) as reporters. The biosensors enable sequence confirmation of the target DNA by hybridization and simple visual detection of the emitted fluorescence under a UV lamp. The ‘diagnostic’ membrane of the biosensor contains a test zone (TZ) and a control zone (CZ). The CZ always fluoresces in order to confirm the proper function of the biosensor. Fluorescence is emitted from the TZ, only when the specific nucleic acid sequence is present. We have developed two general types of QD-based nucleic acid biosensors, namely, Type I and Type II, in which the TZ consists of either immobilized streptavidin (Type I) or immobilized oligodeoxynucleotides (Type II). The control zone consists of immobilized biotinylated albumin. No purification steps are required prior to the application of the DNA sample on the strip. The QD-based nucleic acid biosensors performed accurately and reproducibly when applied to (a) the visual detection of PCR amplification products and (b) visual genotyping of single nucleotide polymorphisms (SNPs) in human genomic DNA from clinical samples. As low as 1.5 fmol of double-stranded DNA were clearly detected by naked eye and the dynamic range extended to 200 fmol. The %CV were estimated to be 4.3–8.2.     

Performance Monitoring and Competence Assessment in Health Services

Health and health service monitoring is among the most promising research area today and the world work towards efficient and cost effective health care. This paper deals with monitoring health service performance using more than one performance outcome variable (multi-attribute processes), which is common in most health services. Although monitoring whether a health service changes or improves over time is important this is well covered in the current literature. Therefore this paper focuses on comparing similar health services in terms of their performance. The proposed procedure is based on an appropriate control chart. The paper deals with firstly the case when no risk adjustment is required because the health services being compared treat the same patient case-mix which does not vary over time. Secondly it deals with comparing health services where risk adjustment is required because the patient case-mix they service do differ because they service either very different geographical locations or service very different demographics of the same population. The technology developed in this paper could be used for example to assess and compare health practitioners’ competence over time, i.e. to decide if two doctors are equivalent in terms of their outcome performances. The waiting time random variable associated with the run length distribution of the control charts (as well as to competence testing) is studied using a Markov Chain embedding technique. Numerical results are provided that exhibit the value of the proposed procedures.     

Elimination of motion,pulsatile flow and cross-talk artifacts using blade sequences in lumbar spine MR imaging

The purpose of this study is to evaluate the ability of T2 turbo spin echo (TSE) axial and sagittal BLADE sequences in reducing or even eliminating motion, pulsatile flow and cross-talk artifacts in lumbar spine MRI examinations. Forty four patients, who had routinely undergone a lumbar spine examination, participated in the study. The following pairs of sequences with and without BLADE were compared: a) T2 TSE Sagittal (SAG) in thirty two cases, and b) T2 TSE Axial (AX) also in thirty two cases. Both quantitative and qualitative analyses were performed based on measurements in different normal anatomical structures and examination of seven characteristics, respectively. The qualitative analysis was performed by experienced radiologists. Also, the presence of image motion, pulsatile flow and cross-talk artifacts was evaluated. Based on the results of the qualitative analysis for the different sequences and anatomical structures, the BLADE sequences were found to be significantly superior to the conventional ones in all the cases. The BLADE sequences eliminated the motion artifacts in all the cases. In our results, it was found that in the examined sequences (sagittal and axial) the differences between the BLADE and conventional sequences regarding the elimination of motion, pulsatile flow and cross-talk artifacts were statistically significant. In all the comparisons, the T2 TSE BLADE sequences were significantly superior to the corresponding conventional sequences regarding the classification of their image quality. In conclusion, this technique appears to be capable of potentially eliminating motion, pulsatile flow and cross-talk artifacts in lumbar spine MR images and producing high quality images in collaborative and non-collaborative patients.     

Asymptotic behaviour of the stochastic Lotka-Volterra model

This paper examines the asymptotic behaviour of the stochastic extension of a fundamentally important population process, namely the Lotka-Volterra model. The stochastic version of this process appears to have far more intriguing properties than its deterministic counterpart. Indeed, the fact that a potential deterministic population explosion can be prevented by the presence of even a tiny amount of environmental noise shows the high level of difference which exists between these two representations.     

Positivity of the weights of interpolatory quadrature formulae with Bernstein–Szegö abscissae

We consider interpolatory quadrature formulae, relative to the Legendre weight function w(t)?=?1 on [???1,1], having as nodes the zeros of any one of the nth degree orthogonal polynomials relative to the Bernstein–Szegö weight functions $$w_{\gamma}^{\left(\pm 1/2\right)}(t)=\frac{\left(1-t^{2}\right)^{\pm 1/2}} {1-\displaystyle\frac{4\gamma}{(1+\gamma)^{2}}t^{2}},\ \ -1<t<1, \ \ -1<\gamma<1.$$ We derive semiexplicit formulae for the quadrature weights and show that the weights are almost all positive; exceptions occur only for the weight corresponding to the node at 0 when n is odd and are carefully identified.     

Adsorption of trypsin on hydrophilic and hydrophobic surfaces

The adsorption of trypsin onto polystyrene and silica surfaces was investigated by reflectometry, spectroscopic methods, and atomic force microscopy (AFM). The affinity of trypsin for the hydrophobic polystyrene surface was higher than that for the hydrophilic silica surface, but steady-state adsorbed amounts were about the same at both surfaces. The conformational characteristics of trypsin immobilized on silica and polystyrene nanospheres were analyzed in situ by circular dichroism and fluorescence spectroscopy. Upon adsorption the trypsin molecules underwent structural changes at the secondary and tertiary level, although the nature of the structural alterations was different for silica and polystyrene surfaces. AFM imaging of trypsin adsorbed on silica showed clustering of enzyme molecules. Rinsing the silica surface resulted in 20% desorption of the originally adsorbed enzyme molecules. Adsorption of trypsin on the surface of polystyrene was almost irreversible with respect to dilution. After adsorption on silica the enzymatic activity of trypsin was 10 times lower, and adsorbed on polystyrene the activity was completely suppressed. The trypsin molecules that were desorbed from the sorbent surfaces by dilution with buffer regained full enzymatic activity.     

Exact Results and Bounds for the Joint Tail and Moments of the Recurrence Times in a Renewal Process

Methodology and Computing in Applied Probability - The best known result about the joint distribution of the backward and forward recurrence times in a renewal process concerns the asymptotic...     

Development and validation of a method for the determination of buprenorphine and norbuprenorphine in breast milk by gas chromatography-mass spectrometry

Buprenorphine (BUP) is used for the maintenance of opioid‐addicted pregnant women. Because BUP and its main metabolite nor‐BUP are excreted into breast milk, a sensitive and specific GC/MS method has been developed, optimized and validated for their determination in breast milk. BUP‐d4 was used as internal standard. The sample preparation includes combination of protein precipitation with solid‐phase extraction and derivatization (acetylation). The absolute recovery for both analytes was found to be higher than 87.3%. The limits of detection and quantification were 0.07 and 0.20 µg/L, respectively. The calibration curves were linear within the dynamic range 0.20–20.0 µg/L, with a correlation coefficient higher than 0.996. Intra‐ and inter‐day accuracies were ranged from ?7.06 to 4.50 and from ?5.88 to 7.00%, respectively, while intra‐ and inter‐day precision were less than 5.7 and 6.1%. The analytes were found to be stable in breast milk at 4°C for one week, at ?20°C for one month, and after three freeze–thaw cycles. The method can be used for the determination of BUP and nor‐BUP in breast milk of BUP‐maintained mothers, in order to calculate the amount of drug that could pass to the newborn via breast milk and to avoid toxic consequences of breastfeeding. Copyright © 2011 John Wiley & Sons, Ltd.     

Dual-cardiac marker capillary waveguide fluoroimmunosensor based on tyramide signal amplification

The early diagnosis of acute myocardial infarction requires the determination of several markers in serum shortly after its incidence. The markers most widely employed are the isoenzyme MB of creatine kinase (CK-MB) and the cardiac troponin I (cTnI). In the present work, a capillary waveguide fluoroimmunosensor for fast and highly sensitive simultaneous determination of these markers in serum samples is demonstrated. The dual-analyte immunosensor was realized using glass capillaries internally modified with an ultrathin poly(dimethylsiloxane) film by creating discrete bands of analyte-specific antibodies. The capillary was then filled with a mixture of sample and biotinylated detection antibodies followed by reaction with streptavidin–horseradish peroxidase and incubation with a fluorescently labeled tyramide derivative to accumulate fluorescent labels onto immunoreaction bands. Upon scanning the capillary with a laser beam, part of the emitted fluorescence is trapped and waveguided through the capillary wall to a photomultiplier placed on one of its ends. The employment of tyramide signal amplification provided detection limits of 0.2 and 0.5 ng/mL for cTnI and CK-MB, respectively, in a total assay time of 30 min compared to 0.8 and 0.6 ng/mL obtained for the corresponding assays when the conventional fluorescent label R-phycoerythrin was used in a 65-min assay. In addition, the proposed immunosensor provided accurate and repeatable measurements (intra-assay and interassay coefficients of variation lower than 10%), and the values determined in serum samples were in good agreement with those obtained with commercially available enzyme immunoassays. Thus, the proposed capillary waveguide fluoroimmunosensor has all the required characteristics for fast and reliable diagnosis of acute myocardial infarction.