Development of a real-time PCR to detect Streptococcus equi subspecies equi.

REASONS FOR PERFORMING STUDY: Infection with Streptococcus equi subspecies equi (S. equi) is endemic in the UK. A proportion of horses serve as long-term carriers and act as a reservoir of infection. Detection of these persistently infected horses is difficult using standard culture techniques owing to a lack of sensitivity and overgrowth by contaminating bacteria. In addition, differentiation of this causative bacterium from the closely related S. equi zooepidemicus has made the development of reliable and accurate diagnostic tests difficult. OBJECTIVE: To develop and validate a sensitive and specific real-time PCR assay to detect S. equi and to compare the results with traditional culture techniques. STUDY DESIGN: Retrospective cross-sectional study. METHODS: The assay was validated using a panel of 92 samples from suspected clinical cases of strangles. These were cultured using microbial techniques and tested using the S. equi real-time PCR. The results of the 2 methods were compared, and the diagnostic sensitivity and specificity of the real-time PCR were calculated. The real-time PCR was tested for cross-reactivity with horse commensal bacteria, and the efficiencies and limits of detection were established. RESULTS: The assay had a diagnostic sensitivity of 95% and specificity of 86%. No cross-reactivity was observed with any of the bacterial species tested, including S. equi zooepidemicus. The assay detected as few as 3 gene copies. CONCLUSION: The assay is fast, sensitive and specific and will detect S. equi DNA directly from a crude extract of clinical material on a swab. POTENTIAL RELEVANCE: This assay could aid in the rapid detection of subclinical shedders of S. equi, enabling quicker treatment and helping to limit the spread of strangles in equine populations.

Proton vibrational dynamics in lithium imide investigated through incoherent inelastic and Compton neutron scattering.

In the present study we report neutron spectroscopic measurements on polycrystalline lithium imide, namely, incoherent inelastic neutron scattering at 20 K, and neutron Compton scattering from 10 K up to room temperature. From the former technique the H-projected density of phonon states up to 100 meV is derived, while the latter works out the spherically averaged single-particle (i.e., H, Li, and N) momentum distributions and, from this, the mean kinetic energies. Only for H at the lowest investigated temperature, non-gaussian components of its momentum distribution are detected. However, these components do not seem directly connected to the system anharmonicity, being fully compatible with the simple N-H bond anisotropy. Neutron data are also complemented by ab initio lattice dynamics simulations, both harmonic and, at room temperature, carried out in the framework of the so-called "quantum colored noise thermostat" method. The single-particle mean kinetic energies in lithium imide as a function of temperature show a quite peculiar behavior at the moment not reproduced by ab initio lattice dynamics methods, at least as far as H and Li are concerned. As matter of fact, neither their low temperature values nor their temperature trends can be precisely explained in terms of standard phonon calculations.

Simultaneous measurement of lithium and fluorine momentum in 7LiF.

We present momentum widths and mean kinetic energies of lithium and fluorine in (7)LiF, as determined simultaneously from deep inelastic neutron scattering (DINS) measurements. Experimental data across a temperature range from 4 to 300 K are presented, and these results compared to those calculated using a quasi-harmonic density-functional approach. In all cases, measured momentum widths are seen to be within approximately 5% of those calculated, despite the very low scattering cross sections of both (7)Li and (19)F. This is the first determination and comparison with theory of such simultaneous measurements for nuclei of mass > 4 amu, and demonstrates the implementation of the DINS method in its current form as a mass-selective neutron spectroscopy.

Proton Momentum Distribution and Diffusion Coefficient in Water: Two Sides of the Same Coin?

Water, the prototype of a liquid to ordinary people, is the most anomalous liquid to physicists, showing regions of the temperature-density (T,ρ) plane where its microscopic structure, diffusion coefficient, and density have anomalous behaviors. Structural anomalies occur over a broad bell-shaped T,ρ region. This region contains, as a matryoshka, two smaller regions, one delimiting dynamical and the other delimiting thermodynamic anomalies. Water anomalous behavior in each of these regions manifests itself as a decrease of order or an increase of the diffusion coefficient upon increasing pressure and as a decrease of density upon cooling. Here, we show that the radial momentum distribution of water protons and their mean kinetic energy have a peculiar, theoretically unpredicted anomaly in the region of dynamical anomalies. This anomaly can be rationalized as due to two distinct "families" of water protons, experiencing quite distinct local environments, leading to an enhancement of the momentum fluctuations along with an increase of kinetic energy.

Neutron Compton scattering investigation of sodium hydride: from bulk material to encapsulated nanoparticulates in amorphous silica gel.

In this study we utilize neutron Compton scattering (NCS) to determine differences in nuclear momentum distributions in NaH, both as bulk material and encapsulated as nanoscale particles (from 20 to 50 nm in diameter) within an amorphous silica-gel matrix (SiGNaH). In addition, elemental Na dispersed in such a matrix is also studied (SiGNa). Data treatment and fitting of experimental spectra yields comparison of the nuclear Compton profiles and radial momentum distributions for the proton in both bulk NaH and nanoscale SiGNaH, with resultant proton kinetic energies being in agreement with previous inelastic neutron studies of bulk NaH. Slight differences in proton radial momentum distributions for bulk and nanoscale systems are witnessed and discussed. The technique of stoichiometric-fixing is applied to the backscattering spectra of each system in order to examine changes in the Na profile width, and NCS is shown to be sensitive to the chemical environment change of this heavier nucleus. Examination of the Si and O profile widths in the gel samples also supports this method.

Changes in the zero-point energy of the protons as the source of the binding energy of water to A-phase DNA.

The measured changes in the zero-point kinetic energy of the protons are entirely responsible for the binding energy of water molecules to A phase DNA at the concentration of 6 water molecules/base pair. The changes in kinetic energy can be expected to be a significant contribution to the energy balance in intracellular biological processes and the properties of nano-confined water. The shape of the momentum distribution in the dehydrated A phase is consistent with coherent delocalization of some of the protons in a double well potential, with a separation of the wells of 0.2 Å.

Proton momentum distribution of liquid water from room temperature to the supercritical phase.

Measurements of the proton momentum distribution n(p) in water from ambient conditions to above the supercritical point are compared with theoretical calculations based on a recently developed polarizable water model. The n(p) along the H-bond direction is narrower in the dense phases, and approaches that of the isolated molecule in the more dilute phases. The theoretical model, which includes only electrostatic interactions, is unable to explain the softening of the local potential experienced by the proton in the dense phases, but it accurately predicts the n(p) for the dilute phases.

Measurement of the 3D Born-Oppenheimer potential of a proton in a hydrogen-bonded system via Deep Inelastic Neutron Scattering: the superprotonic conductor Rb(3)H(SO(4))(2).

We report the first direct measurement of the proton 3-D Born-Oppenheimer potential in any material. The proton potential surfaces in the hydrogen-bonded superprotonic conductor Rb(3)H(SO(4))(2) are extracted from the momentum distribution measured using Deep Inelastic Neutron Scattering (DINS). The potential has a single minimum along the bond direction, which accounts for the absence of the antiferroelectric transition seen in the deuterated material. The measured potential is in qualitative agreement with phenomenological double Morse potentials that have been used to describe hydrogen bonds in other systems.

Measurement of the kinetic energy and lattice constant in hcp solid helium at temperatures 0.07-0.4 K.

The single atom kinetic energy kappa of high purity solid hcp 4He has been measured by neutron Compton scattering, at temperatures between 0.07 and 0.4 K and a pressure of 40 bar. Within statistical error of approximately 2% no change in kappa was observed. The values of kappa at approximately 0.07 K were the same in a single crystal and a polycrystalline sample and were also unaffected (within statistical error) by the addition of 10 ppm of 3He. The lattice constant was also found to be independent of temperature to within 1 part in 2000. These results suggest that the supersolid transition in 4He has a different microscopic origin to the superfluid transition in the liquid.

On the origin of the redshift of the OH stretch in Ice Ih: evidence from the momentum distribution of the protons and the infrared spectral density.

Recent measurements of the momentum distribution in water and ice have shown that the proton is in a considerably softer potential in ice Ih than in water or the free monomer. This is broadly consistent with the large red shift observed in the vibrational spectrum. We show that existing water models, which treat the intramolecular potential as unchanged by the hydrogen bonding are unable to reproduce the momentum distribution. In addition, even if they can substantially explain the red shift they are unable to explain the large increase in intensity observed in the infrared spectrum in going from the monomer to ice Ih. We show that the inclusion of a bond dipole derivative term is essential to explain the observed intensities in the infrared spectrum. Though this term is partially responsible for the softening of the effective potential of the proton we show that best agreement with the observed momentum distribution requires a further softening of the harmonic component of the intramolecular potential. We introduce an efficient normal-mode molecular dynamics algorithm for calculating the momentum distribution with path-integrals.

Anomalous behavior of proton zero point motion in water confined in carbon nanotubes.

The momentum distribution of the protons in ice Ih, ice VI, high density amorphous ice, and water in carbon nanotubes has been measured using deep inelastic neutron scattering. We find that at 5 K the kinetic energy of the protons is 35 meV less than that in ice Ih at the same temperature, and the high momentum tail of the distribution, characteristic of the molecular covalent bond, is not present. We observe a phase transition between 230 and 268 K to a phase that does resemble ice Ih. Although there is yet no model for water that explains the low temperature momentum distribution, our data reveal that the protons in the hydrogen bonds are coherently delocalized and that the low temperature phase is a qualitatively new phase of ice.

Bose-Einstein condensation, phase coherence, and two-fluid behavior in 4He.

It is shown that the assumption that the many particle wave functions describing superfluid 4He have some phase coherence properties of correlated basis functions provides a new explanation of the link between Bose-Einstein condensation and two fluid behavior. This assumption also implies that the condensate fraction is proportional to the superfluid fraction, in agreement with experiment, and provides a simple quantitative explanation of the anomalous reduction in spatial order and liquid density, observed as the temperature is lowered in superfluid 4He.

Direct observation of tunneling in KDP using Neutron Compton scattering.

Neutron Compton scattering measurements presented here of the momentum distribution of hydrogen in KH2PO4 just above and well below the ferroelectric transition temperature are sufficiently sensitive to show clearly that the proton is coherent over both sites in the high temperature phase, a result that invalidates the commonly accepted order-disorder picture of the transition. The Born-Oppenheimer potential for the hydrogen, extracted directly from data for the first time, is consistent with neutron dif-fraction data, and the vibrational spectrum is in substantial agreement with infrared absorption measurements. The measurements are sensitive enough to detect the effect of surrounding ligands on the hydrogen bond, and can be used to study the systematic effect of the variation of these ligands in other hydrogen bonded systems.

Evaluation of the response to a booster dose of hepatitis B vaccine in previously immunized healthcare workers.

INTRODUCTION: Hepatitis B vaccination is recommended for all healthcare workers (HCW) at risk of exposure to infectious body fluids. However, the absolute duration of protection from immunization is unknown. The purpose of this randomized comparison trial was to determine how previously immunized HCW respond to different booster doses of hepatitis B vaccine. METHOD: Adult HCW (n=59) were classified by level of hepatitis B surface antigen (anti-HBs), either <10 milli-International Units per milliliter (mIU/ml) or 10-50 mIU/ml. Participants were then randomized to receive a 2.5 or 10 microg dose of hepatitis B vaccine. Evaluation of anti-HBs levels were conducted 10 to 14 days, one month and one year postbooster. RESULTS AND DISCUSSION: All participants responded to the booster dose with increased anti-HBs levels. At 14 days, mean anti-HBs levels were significantly higher for those with higher levels at baseline (P=0.004) and those receiving the 10 microg dose (P=0.016). At one month, those with higher anti-HBs levels at baseline and those receiving the 10 microg dose were significantly higher (P<0.01 for both). At one year, the increase for the higher dose was no longer statistically significant when examined by itself (P=0.081); statistical significance (P=0.021) was achieved after adjusting for anti-HBs level at baseline. For all participants, the geometric mean anti-HBs level was 2618 mIU/ml at 14 days, 2175 mIU/ml at one month and 88.9 mIU/ml at one year. At all time points the increase in anti-HBs levels represented an increase over the geometric mean baseline level of anti-HBs (7.4 mIU/ml). Hepatitis B immunized adults responded to a booster dose of hepatitis B vaccine from 3 to 13 yr postvaccination series. Data support current recommendations that immunized HCW do not require periodic antibody testing or vaccine boosters.

The physiology behind exercise testing.

Despite significant advances in related technologies, the exercise test remains a useful clinical tool. It can define the limits of performance in an athlete or the functional capabilities of the patient with heart disease. Although virtually all of the body's physiologic and metabolic systems interact in a coordinated fashion to perform a single bout of exhaustive exercise, a major burden falls on the cardiopulmonary system. A good understanding of the basic physiologic responses to acute exercise can assist the clinician in applying the information gained from the test. These responses are influenced by state of health and fitness, age, gender, type of exercise, exercise position, and the environment. This article provides an overview of the physiologic responses to acute exercise, and discusses the many factors that influence these responses.

Bose-einstein condensation and spatial correlations in 4He

A new Monte Carlo method for calculating ground-state properties of liquid 4He is described. It is shown that Bose-Einstein condensation (BEC) implies delocalization of the wave function. It is shown that there is no general connection between the static structure factor and BEC. It is suggested that the observed connection in liquid 4He is due to the creation of spaces in the liquid structure, which are required so that the wave function can delocalize, in the presence of the hard-core interactions. It is shown that this suggestion is quantitatively consistent with observations on liquid 4He.

Hepatitis A vaccine administration: comparison between jet-injector and needle injection.

INTRODUCTION: Type A hepatitis virus (HAV) is a serious health problem throughout the world and can be spread via fecal-oral contact. Both immune globulin and an HAV vaccine provide protection, but the vaccine gives complete protection. Efficacy of methods of vaccination in relation to the formation of anti-HAV antibodies is unclear; thus, this study seeks to determine if significant differences exist between the syringe as compared to the jet injection technique. The purpose of this study was to compare in a randomized trial Biojet jet-injection system to a needle-syringe method. To determine if a significant difference between these two methods in seroconversion rates or geometric mean titers of anti HAV antibody occurs at day 15, 30, and 210 days after vaccination. METHOD: Anti-HAV IgG(-) adult hospital employees were randomized to receive 1440 EL.U of hepatitis a vaccine (HAVRIX(R)) in 2 doses by either needle or jet-injector (Biojector(R)) system at month 0 and 6. HAV seroconversion titer results were measured by the Boehringer-Mannheim method. RESULTS/DISCUSSION: A higher proportion of persons who received HAV vaccine via the Biojector(R) seroconverted with anti-HAV level >/=20 mIU at day 15, 30, and month 7 when compared with a needle injection.Side-effect profiles reported by participants in both methods were below those identified in current published and insert information, but the Biojector(R) had greater local reactivity in all categories when compared to the needle method.