Large clonal expansions of human virus-specific memory cytotoxic T lymphocytes within the CD57+ CD28- CD8+ T-cell population

The proportion of human peripheral blood CD8+ T cells that are CD57+ CD28- is low at birth but increases with age and in individuals infected with human cytomegalovirus (HCMV) or human immunodeficiency virus (HIV). These CD57+ CD28- CD8+ T cells contain large oligoclonal T-cell expansions whose antigen specificity is unknown. We identified clonal expansions of virus-specific memory cytotoxic T-lymphocyte precursors (CTLp) in both healthy carriers of HCMV and in asymptomatic HIV-infected subjects. In each subject, from the T-cell receptor (TCR) beta-chain hypervariable sequence of each immunodominant CTL clone, we designed complementary oligonucleotide probes to quantify the size and phenotypic segregation of individual virus-specific CTL clones in highly purified populations of peripheral blood CD8+ T cells. We found large clonal expansions of virus-specific CTL clonotypes in CD57+ CD28- CD8+ T cells. Using limiting dilution analysis, we found functional peptide-specific CTLp at high frequency in CD57+ CD28- cells. Thus, memory CTL specific for persistent viruses account for many oligoclonal expansions within CD57+ CD28- CD8+ T cells.     

Effect of Velocity Profile Skewing on Blood Velocity and Volume Flow Waveforms Derived From Maximum Doppler Spectral Velocity

Given evidence that fully developed axisymmetric flow may be the exception rather than the rule, even in nominally straight arteries, maximum velocity (V_max) can lie outside the Doppler sample volume (SV). The link between V_max and derived quantities, such as volume flow (Q), may therefore be more complex than commonly thought. We performed idealized virtual Doppler ultrasound on data from image-based computational fluid dynamics (CFD) models of the normal human carotid artery and investigated how velocity profile skewing and choice of sample volume affected V_max waveforms and derived Q variables, considering common assumptions about velocity profile shape (i.e., Poiseuille or Womersley). Severe velocity profile skewing caused substantial errors in V_max waveforms when using a small, centered SV, although peak V_max was reliably detected; errors with a long SV covering the vessel diameter were orientation dependent but lower overall. Cycle-averaged Q calculated from V_max was typically within ±15%, although substantial skewing and use of a small SV caused 10%–25% underestimation. Peak Q derived from Womersley's theory was generally accurate to within ±10%. V_max pulsatility and resistance indexes differed from Q-based values, although the Q-based resistance index could be predicted reliably. Skewing introduced significant error into V_max-derived Q waveforms, particularly during mid-to-late systole. Our findings suggest that errors in the V_max and Q waveforms related to velocity profile skewing and use of a small SV, or orientation-dependent errors for a long SV, could limit their use in wave analysis or for constructing characteristic or patient-specific flow boundary conditions for model studies.     

Compensating for the effect of inlet gas temperature on heated humidifier performance

The humidity output of heated humidifiers may be compromised by inlet gas temperatures exceeding approximately 26 degrees C, with humidity dropping below the recommended levels for intubated patients. A new version of the Fisher & Paykel MR850 humidifier claims to deal with this problem by offering a humidity compensation option. The present study tested this feature by measuring humidity output using the gravimetric method and a hygrometer at different inlet gas temperatures (16.6 degrees C to 40.0 degrees C) with compensation on and off. It was found that the compensation is effective in maintaining humidity levels despite high inlet gas temperatures.     

Mechanisms of a reduced cardiac output and the effects of milrinone and levosimendan in a model of infant cardiopulmonary bypass

OBJECTIVES: A low cardiac output state is an important cause of morbidity after pediatric cardiopulmonary bypass. The objectives of our study were to define the early precipitants of the reduced cardiac output and to investigate the effects on these of milrinone and levosimendan in a model of pediatric cardiopulmonary bypass. DESIGN: Experimental study. SETTING:: Research laboratory at a university-affiliated, tertiary pediatric center. SUBJECTS: Eighteen piglets. INTERVENTIONS: Piglets, instrumented with systemic, pulmonary arterial, and coronary sinus catheters, pulmonary and circumflex arterial flow probes, and a left ventricular conductance-micromanometer-tipped catheter, underwent cardiopulmonary bypass with aortic cross-clamp and cardioplegic arrest. At 120 mins, they were assigned to control, milrinone, or levosimendan groups and studied for a further 120 mins. MEASUREMENTS AND MAIN RESULTS: In controls, between 120 and 240 mins, cardiac output decreased by 15%. Systemic vascular resistance was unchanged, but pulmonary vascular resistance increased by 19%. Systemic arterial elastance increased by 17%, indicating increased afterload. End-systolic elastance was unchanged, and coronary sinus oxygen tension decreased by 4.0 +/- 1.7 mm Hg. In animals receiving milrinone cardiac output was preserved, and in animals receiving levosimendan cardiac output increased by 14%. Both drugs prevented an increase in arterial elastance and pulmonary vascular resistance after cardiopulmonary bypass. Systemic vascular resistance decreased by 31% after levosimendan, and end-systolic elastance increased by 48%, indicating improved contractility. Both agents prevented a decrease in coronary sinus oxygen tension. CONCLUSIONS: Increased afterload, which is not matched by an equivalent elevation in contractility, contributes to the reduced cardiac output early after pediatric cardiopulmonary bypass in this model. This increase is prevented by milrinone and levosimendan. Both agents exert additional beneficial effects on pulmonary vascular resistance and myocardial oxygen balance, although levosimendan has greater inotropic properties.     

Non-linear separation of pressure, velocity and wave intensity into forward and backward components

Separating pressure, flow/velocity and wave intensity signals into forward and backward components provide insights about arterial wave propagation and reflection. A linear wave separation is normally used, but ignores the pressure-dependence of wave speed. While a non-linear separation could incorporate this pressure-dependence, no such method exists for wave intensity decomposition. Moreover, although linear separation errors for pressure (5-10?%) have been quantified previously, errors for velocity and wave intensity have not. Accordingly, we describe a non-linear wave separation technique based on the method of characteristics. Data from a computer model suggest that the percentage linear separation errors for velocity and wave intensity are approximately one-half and twice that for pressure, respectively. Although comparable to measurement uncertainty in many instances, linear separation errors may become more significant: (1) if wave speed varies substantially over the cardiac cycle, e.g. if pulse pressure or vessel compliance is high, (2) if the degree of wave reflection in the arterial system is large, or (3) if the constant wave speed used for the linear separation is closer to the minimum or maximum pressure-dependent value rather than the mean. Consideration of linear separation errors may therefore be important in some physiological settings.     

Linking local knowledge with global action: examining the Global Fund to Fight AIDS, Tuberculosis and Malaria through a knowledge system lens

New global public health institutions are increasingly emphasizing transparency in decision-making, developing-country ownership of projects and programmes, and merit- and performance-based funding. Such principles imply an institutional response to the challenge of bridging the "know-do gap", by basing decisions explicitly on results, evidence and best practice. Using a knowledge systems framework, we examine how the Global Fund to Fight AIDS, Tuberculosis and Malaria has affected the ways in which knowledge is used in efforts to combat these three diseases. We outline the formal knowledge system embedded in current rules and practices associated with the Global Fund's application process, and give three examples that illustrate the complexity of the knowledge system in action: human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS) policy in China; successful applications from Haiti; and responses to changing research on malaria. These examples show that the Global Fund has created strong incentives for knowledge to flow to local implementers, but with little encouragement and few structures for the potentially valuable lessons from implementation to flow back to global best practice or research-based knowledge. The Global Fund could play an influential role in fostering much-needed learning from implementation. We suggest that three initial steps are required to start this process: acknowledging shared responsibility for learning across the knowledge system; analysing the Global Fund's existing data (and refining data collection over time); and supporting recipients and technical partners to invest resources in linking implementation with best practice and research.