Competitive inhibition assay for the detection of progesterone in dairy milk using a fiber optic SPR biosensor.

Analytical methods that are often used for the quantification of progesterone in bovine milk include immunoassays and chromatographic techniques. Depending on the selected method, the main disadvantages are the cost, time-to-result, labor intensity and usability as an automated at-line device. This paper reports for the first time on a robust and practical method to quantify small molecules, such as progesterone, in complex biological samples using an automated fiber optic surface plasmon resonance (FO-SPR) biosensor. A FO-SPR competitive inhibition assay was developed to determine biologically relevant concentrations of progesterone in bovine milk (1-10 ng/mL), after optimizing the immobilization of progesterone-bovine serum albumin (P4-BSA) conjugate, the specific detection with anti-progesterone antibody and the signal amplification with goat anti-mouse gold nanoparticles (GAM-Au NPs). The progesterone was detected in a bovine milk sample with minimal sample preparation, namely ½ dilution of the sample. Furthermore, the developed bioassay was benchmarked against a commercially available ELISA, showing excellent agreement (R2 = 0.95). Therefore, it is concluded that the automated FO-SPR platform can combine the advantages of the different existing methods for quantification of progesterone: sensitivity, accuracy, cost, time-to-result and ease-of-use.

Real-time PCR melting analysis with fiber optic SPR enables multiplex DNA identification of bacteria.

A fiber optic surface plasmon resonance (FO-SPR) technology was developed that enables simultaneous quantification and identification of multiple DNA targets on the same platform. The bioassay was based on the hybridization/melting of DNA-coated Au nanoparticles on the FO-SPR sensor when targets are present. The multiplex concept was successfully demonstrated on two related bacteria and for detection of multiple mutations in sequences. In conclusion, FO-SPR technology shows a great potential as a next generation in vitro diagnostics tool.

Potentiometric sensors doped with biomolecules as a new approach to small molecule/biomolecule binding kinetics analysis.

The most successful binding kinetics analysis systems at this moment include surface plasmon resonance (SPR), quartz microcrystal balance (QMB) and surface acoustic wave (SAW). Although these are powerful methods, they generally are complex, expensive and require the use of monolayers. Here, we report on potentiometric sensors as an inexpensive and simple alternative to do binding kinetics analysis between small molecules in solution and biomolecules (covalently) attached in a biopolymer sensor coating layer. As an example, dopamine and an anti-dopamine aptamer were used as the small molecule and the biomolecule respectively. Binding between both follows a Langmuir adsorption type model and creates a surface potential. The system operates in Flow Injection Analysis mode (FIA). Besides being an interesting new binding kinetics tool, the approach allows systematic design of potentiometric biosensors (in the present study a dopamine sensor), and gives new insights into the functioning of ion-selective electrodes (ISE's).

Robust quantum control by a single-shot shaped pulse.

Considering the problem of the control of a two-state quantum system by an external field, we establish a general and versatile method allowing the derivation of smooth pulses which feature the properties of high fidelity, robustness, and low area. Such shaped pulses can be interpreted as a single-shot generalization of the composite pulse-sequence technique with a time-dependent phase.

Use of potentiometric detection in (ultra) high performance liquid chromatography and modelling with adsorption/desorption binding kinetics.

Observation of a potentiometric sensor's response behaviour after injection in flow injection analysis at different concentrations allowed studying "on" and "off" kinetics of the analyte's adsorption/diffusion behaviour. The alkaloid metergoline was mostly used as an example. k(on) and k(off) rate constant values were measured, and the association constant K(ass), and ΔG values of the analyte-surface interaction were calculated with an adsorption-based model which proved to be fully applicable. k(on) increased by decreasing the sensor dimensions, while koff was unaffected by miniaturization. Increasing acetonitrile concentrations in the running buffer increased k(off), while k(on) was unaffected. The experimentally determined ΔG values of the analyte-surface interaction showed a linear relation to the response of the sensor, in mV. This knowledge was applied to optimize the potentiometric detection of plant alkaloids in (U)HPLC. Sub-micromolar detection limits were obtained with the potentiometric detector/(U)HPLC combination. This is the first time that the response rates and the response itself can be modelled accurately for coated wire potentiometric sensors, and it is the first application of a potentiometric detector in UPLC.

Use of potentiometric sensors to study (bio)molecular interactions.

Potentiometric sensors were used to study molecular interactions in liquid environments with sensorgram methodology. This is demonstrated with a lipophilic rubber-based and a collagen-based hydrogel sensor coating. The investigated molecules were promazine and tartaric acid, respectively. The sensors were placed in a hydrodynamic wall-jet system for the recording of sensorgrams. Millivolt sensor responses were first converted to a signal, expressing the concentration of adsorbed organic ions. Using a linearization method, a pseudo-first order-kinetic model of adsorption was shown to fit the experimental results perfectly. K(assoc), k(on), and k(off) values were calculated. The technique can be used over 4 decades of concentration, and it is very sensitive to low-MW compounds as well as to multiply charged large biomolecules. This study is the first to demonstrate the application of potentiometric sensors as an alternative and complement to surface plasmon resonance methods.

The effects of long-term noninvasive ventilation in hypercapnic COPD patients: a randomized controlled pilot study.

INTRODUCTION: Noninvasive ventilation (NIV) is a well-established treatment for acute-on- chronic respiratory failure in hypercapnic COPD patients. Less is known about the effects of a long-term treatment with NIV in hypercapnic COPD patients and about the factors that may predict response in terms of improved oxygenation and lowered CO(2) retention. METHODS: In this study, we randomized 15 patients to a routine pharmacological treatment (n = 5, age 66 [standard deviation ± 6] years, FEV(1) 30.5 [±5.1] %pred, PaO(2) 65 [±6] mmHg, PaCO(2) 52.4 [±6.0] mmHg) or to a routine treatment and NIV (using the Synchrony BiPAP device [Respironics, Inc, Murrsville, PA]) (n = 10, age 65 [±7] years, FEV(1) 29.5 [±9.0] %pred, PaO(2) 59 [±13] mmHg, PaCO(2) 55.4 [±7.7] mmHg) for 6 months. We looked at arterial blood gasses, lung function parameters and performed a low-dose computed tomography of the thorax, which was later used for segmentation (providing lobe and airway volumes, iVlobe and iVaw) and post-processing with computer methods (providing airway resistance, iRaw) giving overall a functional image of the separate airways and lobes. RESULTS: In both groups there was a nonsignificant change in FEV(1) (NIV group 29.5 [9.0] to 38.5 [14.6] %pred, control group 30.5 [5.1] to 36.8 [8.7] mmHg). PaCO(2) dropped significantly only in the NIV group (NIV: 55.4 [7.7] → 44.5 [4.70], P = 0.0076; control: 52.4 [6.0] → 47.6 [8.2], NS). Patients actively treated with NIV developed a more inhomogeneous redistribution of mass flow than control patients. Subsequent analysis indicated that in NIV-treated patients that improve their blood gases, mass flow was also redistributed towards areas with higher vessel density and less emphysema, indicating that flow was redistributed towards areas with better perfusion. There was a highly significant correlation between the % increase in mass flow towards lobes with a blood vessel density of >9% and the increase in PaO(2). Improved ventilation-perfusion match and recruitment of previously occluded small airways can explain the improvement in blood gases. CONCLUSION: We can conclude that in hypercapnic COPD patients treated with long-term NIV over 6 months, a mass flow redistribution occurs, providing a better ventilation-perfusion match and hence better blood gases and lung function. Control patients improve homogeneously in iVaw and iRaw, without improvement in gas exchange since there is no improved ventilation/perfusion ratio or increased alveolar ventilation. These differences in response can be detected through functional imaging, which gives a more detailed report on regional lung volumes and resistances than classical lung function tests do. Possibly only patients with localized small airway disease are good candidates for long-term NIV treatment. To confirm this and to see if better arterial blood gases also lead to better health related quality of life and longer survival, we have to study a larger population.

Pulmonary rehabilitation and non-invasive ventilation in COPD.

A multidisciplinary pulmonary rehabilitation program has become an important part of the treatment of chronic obstructive pulmonary disease. It can improve both exercise tolerance and health related quality of life in these patients. Exercise training has to be included for the program to be successful. The intensity of the training is of great importance: there is more physiological benefit in high-intensity training, compared to moderate-intensity training. High-intensity training results in reduced levels of blood lactate and pulmonary ventilation at a given heavy work rate. High-intensity training is limited in COPD patients because of exercise-induced dyspnoea. Flow limitation, as a consequence of increased ventilatory demands of exercise, causes a breathing pattern with greater demands on their inspiratory muscles: this results in a pattern of low tidal volume and high-frequency breathing. Increased inspiratory muscle work causes dyspnoea and limitation in exercise intensity. Artificial ventilatory assistance could improve exercise tolerance and hence help severe COPD patients to achieve a higher level of training. It could help to unload and assist the overburdened ventilatory muscles and give a possibility for higher levels of exercise intensity. In this review article we will discuss the effectiveness and feasibility of training with ventilatory aids.

Transitions in the communication capacity of dissipative qubit channels.

The information transmission is studied for quantum channels in which the noise includes dissipative effects, more specifically, nonunitality. Noise is usually a nuisance but can sometimes be helpful. For these channels, the communication capacity is shown to increase with the dissipative component of the noise and may exhibit transitions beyond which it increases faster. The optimal states are constructed analytically as well as the pertaining "phase" diagram.

Adiabatic quantum search scheme with atoms in a cavity driven by lasers.

We propose an implementation of the quantum search algorithm of a marked item in an unsorted list of N items by adiabatic passage in a cavity-laser-atom system. We use an ensemble of N identical three-level atoms trapped in a single-mode cavity and driven by two lasers. In each atom, the same level represents a database entry. One of the atoms is marked by having an energy gap between its two ground states. Appropriate time delays between the two laser pulses allow one to populate the marked state starting from an initial entangled state within a decoherence-free adiabatic subspace. The time to achieve such a process is shown to exhibit the square root N Grover speedup.

Intermittent positive airway pressure by nasal mask as a treatment for respiratory insufficiency in a patient with Charcot-Marie-Tooth disease.

Charcot-Marie-Tooth disease (CMT) is a slowly progressive hereditary neuropathy characterised by degeneration of motor and sensory peripheral nerves resulting in distal muscle weakness with atrophy and sensory impairment. We report a 35-year-old woman with CMT presenting with respiratory failure due to a pneumonia, sputum impaction and insufficient cough reflex. After recovery, we diagnosed a very severe restrictive lung function disturbance caused by muscle weakness and a possible coexistent unilateral diaphragm paralysis. A very severe REM (Rapid Eye Movement Sleep) related sleep hypopnea syndrome was successfully treated with Nasal Intermittent Positive Pressure Ventilation (NIPPV).

Field-free two-direction alignment alternation of linear molecules by elliptic laser pulses.

We show that a linear molecule subjected to a short specific elliptically polarized laser field yields post-pulse revivals exhibiting alignment alternatively located along the orthogonal axis and the major axis of the ellipse. The effect is experimentally demonstrated by measuring the optical Kerr effect along two different axes. The conditions ensuring an optimal field-free alternation of high alignments along both directions are derived.

Efficient and long-lived field-free orientation of molecules by a single hybrid short pulse.

We show that a combination of a half-cycle pulse and a short nonresonant laser pulse produces a strongly enhanced postpulse orientation. Robust transients that display both efficient and long-lived orientation are obtained. The mechanism is analyzed in terms of optimal oriented target states in finite Hilbert subspaces and shows that hybrid pulses can prove useful for other control issues.

Properties of stationary nonequilibrium states in the thermostatted periodic Lorentz gas: the multiparticle system.

We study the stationary nonequilibrium states of N-point particles moving under the influence of an electric field E among fixed obstacles (disk) in a two-dimensional torus. The total kinetic energy of the system is kept constant through a Gaussian thermostat that produces a velocity dependent mean field interaction between the particles. The current and the particle distribution functions are obtained numerically and compared for small /E/ with analytic solutions of a Boltzmann-type equation obtained by treating the collisions with the obstacles as random independent scatterings. The agreement is surprisingly good for both small and large N. The latter system in turn agrees with a self-consistent one-particle evolution expected to hold in the N-->infinity limit.

Continued fraction representation of temporal multiscaling in turbulence.

It was shown recently that the anomalous scaling of simultaneous correlation functions in turbulence is intimately related to the breaking of temporal scale invariance, which is equivalent to the appearance of infinitely many times scales in the time dependence of time-correlation functions. In this paper we derive a continued fraction representation of turbulent time correlation functions which is exact and in which the multiplicity of time scales is explicit. We demonstrate that this form yields precisely the same scaling laws for time derivatives and time integrals as the "multi-fractal" representation that was used before. Truncating the continued fraction representation yields the "best" estimates of time correlation functions if the given information is limited to the scaling exponents of the simultaneous correlation functions up to a certain, finite order. It is worth noting that the derivation of a continued fraction representation obtained here for a time evolution operator which is not Hermitian or anti-Hermitian may be of independent interest.

Probabilistic and thermodynamic aspects of dynamical systems.

The probabilistic approach to dynamical systems giving rise to irreversible behavior at the macroscopic, mesoscopic, and microscopic levels of description is outlined. Signatures of the complexity of the underlying dynamics on the spectral properties of the Liouville, Frobenius-Perron, and Fokker-Planck operators are identified. Entropy and entropy production-like quantities are introduced and the connection between their properties in nonequilibrium steady states and the characteristics of the dynamics in phase space are explored. (c) 1998 American Institute of Physics.

Fluorescence quenching with lindane in small unilamellar L,alpha-dimyristoylphosphatidylcholine vesicles.

The lateral mobility and lipid-water partition of the pesticide lindane was studied by fluorescence quenching of N-isopropylcarbazole (NIPC) and L,alpha-palmitoyl-beta-(N-carbazolyl) undecanoylphosphatidylcholine (PCUPC) in liposomes of dimyristoylphosphatidylcholine at 50 degrees C. In isotropic solvents the quenching reaction was highly inefficient. A scheme for dynamic quenching, in which the monomolecular quenching rate constant is small, was valid. In lipid bilayers the same scheme was applied to describe the quenching results but the rate constant of the back-reaction of the excited complex to quencher and excited probe was of comparable magnitude to the monomolecular quenching rate constant. This phenomenon results in biexponential decays of the fluorescent probe in the presence of quencher. All the rate constants of the scheme could be determined. Stern-Volmer plots at different membrane concentrations were obtained from fluorescence intensity and decay time measurements. From these plots the true bimolecular quenching rate constant, Kq, and the rate constant for lateral diffusion, kd, were determined: kq[NIPC] = 3.2 +/- 0.5 x 10(8) M-1 s-1, kq[PCUPC] = 1.9 +/- 0.4 x 10(8) M-1 s-1, kd[NIPC] = 6.6 +/- 0.8 x 10(8) M-1 s-1. The smaller value of kq compared to kd for the quenching reaction of NIPC with lindane indicates that this quenching reaction is not diffusion controlled. The lateral diffusion coefficient D of lindane was found to be 1.7 +/- 0.2 x 10(-6) cm2/s in dimyristoylphosphatidylcholine vesicles at 50 degrees C. The partition coefficient of lindane in these lipid bilayers is very high (greater than 2000).

Fluorescence decay of pyrene in small and large unilamellar L, alpha-dipalmitoylphosphatidylcholine vesicles above and below the phase transition temperature.

The fluorescence decays of pyrene in small and large unilamellar L, alpha-dipalmitoylphosphatidylcholine vesicles have been investigated as a function of probe concentration and temperature. When the molar ratio of pyrene to phospholipid equals 1:3000, no excimer emission is observed and the fluorescence decays are mono-exponential. When this ratio is equal to or higher than 1:120, excimer formation is observed. Above the phase transition temperature the observed fluorescence decays of monomer and excimer can be adequately described by a bi-exponential function. The monomer decays can be equally well fitted to a decay law which takes into account a time-dependence in the probe diffusion rate constant. The fluorescence decay kinetics are compatible with the excimer formation scheme which is valid in an isotropic medium. The excimer lifetime and the (apparent) rate constant of excimer formation have been determined as a function of probe concentration at different temperatures above the phase transition temperature. The activation energy of excimer formation is found to be 29.4 +/- 1.3 kJ/mol. In small unilamellar vesicles the diffusion constant associated with the pyrene excimer formation process varies from 8.0 X 10(-7) cm2/s at 40 degrees C to 2.2 X 10(-6) cm2/s at 70 degrees C. Below the phase transition temperature the monomer decays can be described by a decay law which takes into account a time dependence of the rate constant of excimer formation. The lateral diffusion coefficient of pyrene calculated from the decay fitting parameters of the monomer region varies from 4.0 X 10(-9) cm2/s at 20 degrees C to 7.9 X 10(-8) cm2/s at 35 degrees C. No significant difference could be observed between the pyrene fluorescence decay kinetics in small and large unilamellar vesicles.