Reagent integration and controlled release for multiplexed nucleic acid testing in disposable thermoplastic 2D microwell arrays.

The seamless integration of reagents into microfluidic devices can serve to significantly reduce assay complexity and cost for disposable diagnostics. In this work, the integration of multiplexed reagents into thermoplastic 2D microwell arrays is demonstrated using a scalable pin spotting technique. Using a simple and low-cost narrow-bore capillary spotting pin, high resolution deposition of concentrated reagents within the arrays of enclosed nanoliter-scale wells is achieved. The pin spotting method is further employed to encapsulate the deposited reagents with a chemically modified wax layer that serves to prevent disruption of the dried assay components during sample introduction through a shared microchannel, while also enabling temperature-controlled release after sample filling is complete. This approach supports the arbitrary patterning and release of different reagents within individual wells without crosstalk for multiplexed analyses. The performance of the in-well spotting technique is characterized using on-chip rolling circle amplification to evaluate its potential for nucleic acid-based diagnostics.

Community engagement interventions for communicable disease control in low- and lower- middle-income countries: evidence from a review of systematic reviews.

BACKGROUND: Community engagement (CE) interventions include a range of approaches to involve communities in the improvement of their health and wellbeing. Working with communities defined by location or some other shared interest, these interventions may be important in assisting equity and reach of communicable disease control (CDC) in low and lower-middle income countries (LLMIC). We conducted an umbrella review to identify approaches to CE in communicable disease control, effectiveness of these approaches, mechanisms and factors influencing success. METHODS: We included systematic reviews that: i) focussed on CE interventions; ii) involved adult community members; iii) included outcomes relevant to communicable diseases in LLMIC; iv) were written in English. Quantitative results were extracted and synthesised narratively. A qualitative synthesis process enabled identification of mechanisms of effect and influencing factors. We followed guidance from the Joanna Briggs Institute, assessed quality with the DARE tool and reported according to standard systematic review methodology. RESULTS: Thirteen systematic reviews of medium-to-high quality were identified between June and July 2017. Reviews covered the following outcomes: HIV and STIs (6); malaria (2); TB (1); child and maternal health (3) and mixed (1). Approaches included: CE through peer education and community health workers, community empowerment interventions and more general community participation or mobilisation. Techniques included sensitisation with the community and involvement in the identification of resources, intervention development and delivery. Evidence of effectiveness of CE on health outcomes was mixed and quality of primary studies variable. We found: i) significantly reduced neonatal mortality following women's participatory learning and action groups; ii) significant reductions in HIV and other STIs with empowerment and mobilisation interventions with marginalised groups; iii) significant reductions in malaria incidence or prevalence in a small number of primary studies; iv) significant reductions in infant diarrhoea following community health worker interventions. Mechanisms of impact commonly occurred through social and behavioural processes, particularly: changing social norms, increasing social cohesion and social capacity. Factors influencing effectiveness of CE interventions included extent of population coverage, shared leadership and community control over outcomes. CONCLUSION: Community engagement interventions may be effective in supporting CDC in LLMIC. Careful design of CE interventions appropriate to context, disease and community is vital.

Generalized quantum cumulant dynamics.

A means of unifying some semiclassical models of computational chemistry is presented; these include quantized Hamiltonian dynamics, quantal cumulant dynamics, and semiclassical Moyal dynamics (SMD). A general method for creating the infinite hierarchy of operator dynamics in the Heisenberg picture is derived together with a general method for truncation (or closure) of that series, and in addition, we provide a simple link to the phase space methods of SMD. Operator equations of arbitrary order may be created readily, avoiding the tedious algebra identified previously. Truncation is based on a simple recurrence formula which is related to, but avoids the more complex contractions of, Wick's theorem. This generalized method is validated against a number of trial problems considered using the previous methods. We also touch on some of the limitations involved using such methods, noting, in particular, that any truncation will lead to a state which is in some sense unphysical. Finally, we briefly introduce our quantum algebra package QuantAL which provides an automated method for the generation of the required equation set, the initial conditions for all variables from any start, and all the higher order approximations necessary for truncation of the series, at essentially arbitrary order.

Spatially resolved single photon detection with a quantum sensor array.

We propose a method of resolving a spatially coherent signal, which contains on average just a single photon, against the background of local noise at the same frequency. The method is based on detecting the signal simultaneously in several points more than a wavelength apart through the entangling interaction of the incoming photon with the quantum metamaterial sensor array. The interaction produces the spatially correlated quantum state of the sensor array, characterised by a collective observable (e.g., total magnetic moment), which is read out using a quantum nondemolition measurement. We show that the effects of local noise (e.g., fluctuations affecting the elements of the array) are suppressed relative to the signal from the spatially coherent field of the incoming photon as , where N is the number of array elements. The realisation of this approach in the microwave range would be especially useful and is within the reach of current experimental techniques.

Impact of donor left ventricular hypertrophy on survival after heart transplant.

Left ventricular hypertrophy (LVH) of the donor heart is believed to increase the risk of allograft failure after transplant. However this effect is not well quantified, with variable findings from single-center studies. The United Network for Organ Sharing database was used to analyze the effect of donor LVH on recipient survival. Three cohorts, selected in accordance with the American Society of Echocardiography guidelines, were examined: recipients of allografts without LVH (<1.1 cm), with mild LVH (1.1-1.3 cm) and with moderate-severe LVH (≥ 1.4 cm). The study group included 2626 patients with follow-up of up to 3.3 years. Mild LVH was present in 38% and moderate-severe LVH in 5.6% of allografts. Predictors of mortality included a number of donor and recipient characteristics, but not LVH. However, a subgroup analysis showed an increased risk of death in recipients of allografts with LVH and donor age >55 years, and in recipients of allografts with LVH and ischemic time ≥ 4 h. In the contemporary era, close to half of all transplanted allografts demonstrate LVH, and survival of these recipients is similar to those without LVH. However, the use of allografts with LVH in association with other high-risk characteristics may result in increased mortality.

Effect of blood product transfusion-induced tolerance on incidence of cardiac allograft rejection.

BACKGROUND: Blood product transfusion has been successfully used in solid-organ transplantation to induce tolerance. Whether a similar protective effect of blood product transfusion exists in heart transplantation is controversial. OBJECTIVE: To investigate the effect of cellular blood product transfusion within 2 weeks posttransplantation on the incidence of cellular and antibody-mediated rejection. PATIENTS AND METHODS: Patients were grouped on the basis of number of blood transfusions; group 1 received no transfusions, and groups 2, 3, and 4 each received an incremental number of transfusion units. All endomyocardial biopsy samples were routinely studied using immunofluorescence in the first 12 weeks posttransplantation. RESULTS: Baseline characteristics including age, sex, body mass index, history of diabetes, donor characteristics, and pretransplantation laboratory values were similar except that group 4 had a higher rate of previous sternotomy and longer ischemic time during transplantation. Approximately 9200 endomyocardial biopsy samples composed the data. Short- and long-term freedom from the International Society for Heart & Lung Transplantation grade 3A or higher cellular rejection and from antibody-mediated rejection were comparable between groups. CONCLUSIONS: Blood transfusions within the first 2 weeks post-transplantation do not seem to confer any protective effect against posttransplantation cellular rejection or antibody- mediated rejection. Whether other unmeasured confounding factors mask their effect requires further prospective studies.

Recovery of classical chaoticlike behavior in a conservative quantum three-body problem.

Recovering trajectories of quantum systems whose classical counterparts display chaotic behavior has been a subject that has received a lot of interest over the last decade. However, most of these studies have focused on driven and dissipative systems. The relevance and impact of chaotic-like phenomena to quantum systems has been highlighted in recent studies which have shown that quantum chaos is significant in some aspects of quantum computation and information processing. In this paper we study a three-body system comprising of identical particles arranged so that the system's classical trajectories exhibit Hamiltonian chaos. Here we show that it is possible to recover very nearly classical-like, conservative, chaotic trajectories from such a system through an unravelling of the master equation. First, this is done through continuous measurement of the position of each system. Second, and perhaps somewhat surprisingly, we demonstrate that we still obtain a very good match between the classical and quantum dynamics by weakly measuring the position of only one of the oscillators.

Control of multilevel voltage states in a hysteretic superconducting-quantum-interference-device ring-resonator system.

In this paper we study numerical solutions to the quasiclassical equations of motion for a superconducting-quantum-interference device ring-radio frequency (rf) resonator system in the regime where the ring is highly hysteretic. In line with experiment, we show that for a suitable choice of ring circuit parameters the solutions to these equations of motion comprise sets of levels in the rf voltage-current dynamics of the coupled system. We further demonstrate that transitions, both up and down, between these levels can be controlled by voltage pulses applied to the system, thus opening up the possibility of high order (e.g., 10 state), multilevel logic and memory.

A compact electric potential sensor array for the acquisition and reconstruction of the 7-lead electrocardiogram without electrical charge contact with the skin.

Conventional electrocardiogram (ECG) systems make use of separate electrical connections to the arms and legs. These use a 'long baseline' for the voltage reference potential which in the case of precordial ECG leads is provided using a Wilson central terminal (WCT) wiring configuration. The aims of this project were (a) to construct compact, non-invasive surface ECG sensor arrays which would operate without the need for a WCT reference, (b) to obtain high quality precordial ECGs showing fine differences in ECG detail between small adjacent areas of the chest and (c) to reconstruct, from a compact array of four sensors, ECGs which closely match to the conventional 7-lead ECG system, but without the need for multiple wires and long baselines. In this paper, we describe two sensor array configurations which have been constructed using electric potential sensors (EPSs). We show high quality precordial ECGs obtained from small areas of the surface of the chest and show the different angular vectors (leads) in the frontal cardiac plane constructed using signals from the array elements. We suggest that these ECG arrays, which are simple to apply, should prove to be a valuable tool in providing useful information about the state of the heart.

Signatures of chaoticlike and nonchaoticlike behavior in a nonlinear quantum oscillator through photon detection.

The driven nonlinear Duffing oscillator is a very good, and standard, example of a quantum mechanical system from which classical-like orbits can be recovered from unravelings of the master equation. In order to generate such trajectories in the phase space of this oscillator, in this paper we use the quantum jump unraveling together with a suitable application of the correspondence principle. We analyze the measured readout by considering the power spectra of photon counts produced by the quantum jumps. Here we show that localization of the wave packet from the measurement of the oscillator by the photon detector produces a concomitant structure in the power spectra of the measured output. Furthermore, we demonstrate that this spectral analysis can be used to distinguish between different modes of the underlying dynamics of the oscillator.

Pinch resonances in a radio-frequency-driven superconducting-quantum-interference-device ring-resonator system.

In this paper we present experimental data on the frequency domain response of a superconducting-quantum-interference-device ring (a Josephson weak link enclosed by a thick superconducting ring) coupled to a radio frequency tank circuit resonator. We show that with the ring weakly hysteretic the resonance line shape of this coupled system can display opposed fold bifurcations that appear to touch (pinch off). We demonstrate that for appropriate circuit parameters these pinch off line shapes exist as solutions of the nonlinear equations of motion for the system.

Exercise in leisure time: coronary attack and death rates.

Nine thousand three hundred and seventy six male civil servants, aged 45-64 at entry, with no clinical history of coronary heart disease, were followed for a mean period of 9 years and 4 months during which 474 experienced a coronary attack. The 9% of men who reported that they often participated in vigorous sports or did considerable amounts of cycling or rated the pace of their regular walking as fast (over 4 mph, 6.4 km/h) experienced less than half the non-fatal and fatal coronary heart disease of the other men. In addition, entrants aged 55-64 who reported the next lower degree of this vigorous aerobic exercise had rates less than two thirds of the remainder; entrants of 45-54 did not show such an effect. When these forms of exercise were not vigorous they were no protection against the disease, nor were other forms of exercise or high totals of physical activity per se. A history of vigorous sports in the past was not protective. Indications in these men are of protection by specific exercise: vigorous, aerobic, with a threshold of intensity for benefit and "dose response" above this threshold, exercise that has to be habitual, and continuing, which suggests that protection is against the acute phases of the disease. Those men who took vigorous aerobic exercise were demonstrably a favourably "selected" group; they suffered less of the disease, however, whether at low risk or high by the several risk factors that were studied. Men with exercise-related reduction in coronary heart disease also had lower death rates from the total of other causes, and so lower total death rates than the rest of the men.

Vigorous exercise in leisure-time: protection against coronary heart disease.

1138 first clinical episodes of coronary heart disease (CHD) in 17 944 middle-aged male office workers in the Civil Service are reported. Men who engaged in vigorous sports, keep-fit, and the like during an initial survey in 1968-70 had an incidence of CHD in the next 8 1/2 years somewhat less than half that of their colleagues who recorded no vigorous exercise. The CHD rates of men who took such vigorous exercise were lower in both fatal and non-fatal clinical manifestations, though more so in fatal; throughout the age-range studied, though more striking in later middle age and early old age; and in all other sub-groups examined, including men with a family history of CHD, the obese, the short of stature, cigarette smokers, and men with severe hypertension and subclinical angina, as well as in those in favourable situations for CHD or neutral. The generality of the advantage suggests that vigorous exercise is a natural defence of the body, with a protective effect on the ageing heart against ischaemia and its consequences.

Purification and partial characterization of an alpha-chymotrypsin-like protease of rat peritoneal mast cells.

An alpha-chymotrypsin-like enzyme was isolated from mast cells of the rat peritoneal cavity by extraction with 0.8 M potassium phosphate, 2 per cent protamine sulfate followed by affinity chromatography on hen ovoinhibitor-agarose and adsorption on barium sulfate. This procedure yielded over 9 mg of protease from the peritoneal lavage fluid of 100 rats, equivalent to 44 per cent of the initial activity. The purified protein was homogeneous as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, analytical isoelectric focusing, and amino-terminal sequence analysis. The protease contains no covalently bound carbohydrate and has a molecular weight of approximately 26,000. The enzyme molecule is a single polypeptide chain with an amino-terminal sequence homologous to that of the B chain of bovine alpha-chymotrypsin. The kinetic parameters, Km and kcat, for the hydrolysis of N-benzoyl-L-tyrosine ethyl ester were determined at pH 8.0 and 25 degrees C as 1.1 X 10(-3) M and 84 sec-1, respectively. The value of the second-order rate constant for inactivation of mast cell protease by diisopropylphosphofluoridate was 300 times lower than for bovine alpha-chymotrypsin.

A major serine protease in rat skeletal muscle: evidence for its mast cell origin.

The physical, chemical, and immunologic properties of a protease from rat skeletal muscle, proposed to function in the degradation of certain intracellular enzymes, are identical to those of a chymotrypsin-like serine protease isolated from peritoneal mast cells. The results of polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate and 8 M urea indicate that the two rat proteases have identical mobilities corresponding to a molecular weight of 26,000. The relative amino acid compositions of the proteases are nearly identical. Immunodiffusion tests for crossreaction between the muscle protease and antisera directed toward mast cell protease indicate that the former is immunologically identical to mast cell protease. The first 35 amino-terminal residues of the two enzymes are identical and indicate homology of these proteins to other mammalian serine proteases. The sequence analysis of the protease from muscle was extended for an additional 16 positions, and comparison of this amino-terminal sequence with that of a similar enzyme from small intestine showed approximately 75% sequence identity. In contrast, only 40% of the residues in this region of bovine chymotrypsin A were found at corresponding loci in rat muscle protease. It is concluded that the protease from muscle or mast cells is closely related to the enzyme from small intestine which recently was localized in the "atypical" mast cells of gut mucosa [Woodbury, R. G., Gruzenski, G. M. & Lagunoff, D. (1978) Proc. Natl. Acad. Sci. USA 75, 2785-2789].