Sputum changes associated with therapy for endobronchial exacerbation in cystic fibrosis

We sought to define objective indicators of the resolution of Pseudomonas aeruginosa endobronchial infection in patients with cystic fibrosis. We prospectively studied 75 patients admitted for treatment of a pulmonary exacerbation and quantitated sputum bacterial density, DNA content, and the concentration of albumin and total protein in sputum, and compared these values with clinical evaluation. Eleven of the 75 patients had systemic signs, fever, and leukocytosis, which we arbitrarily defined as due to endobronchial infection. At the end of hospitalization, these 11 patients were afebrile, had peripheral leukocyte counts in the normal range, and were judged improved. Sputum P. aeruginosa density, DNA content, and total protein content on admission were similar in the two illness groups. Hospitalization and parenteral antibiotic administration for an average of 14.6 days were associated with improved pulmonary function in all 75 subjects (P values for forced vital capacity, forced expiratory volume at 1 second, and peak expiratory flow rate were all less than 0.001). With improvement, there was a decrease in sputum P. aeruginosa density (mean of both groups decreased from 10(7.80) CFU/g on admission to 10(5.96) CFU/g; P less than 0.001), and a decreased DNA concentration (overall mean 4.73 +/- 4.75 on admission to 2.76 +/- 2.49 mg/g; P less than 0.002). The decrease in sputum total protein concentration for both groups was not significant (overall mean 60.5 +/- 48.4 to 43.9 +/- 38.2 mg/g; P = 0.06). Sputum albumin concentrations did not change in either group. We conclude that in cystic fibrosis subjects with a pulmonary exacerbation, bacterial density, sputum DNA and protein content decrease with hospitalization and parenteral antibiotic therapy. At the end of treatment, these indices of sputum infection and inflammation correlate with improved pulmonary function and clinical improvement. These changes are independent of the presence or absence of fever on admission.     

Evaluating healing for cancer in a community setting from the perspective of clients and healers: a pilot study

The real-life practice of ‘healing’ for cancer in the community as perceived by clients and healers was investigated in a multi-method pilot study. Fifteen clients received six weekly healing sessions. Pre- and post-changes in perception towards well-being and client experience were assessed by EuroQol (EQ-5D), measure yourself concerns and well-being (MYCaW) and a client satisfaction tool. Qualitative methods, including focus groups, explored the perceived effects of healing in more depth and the participants’ experience of taking part in research. The study was not designed to test the effect of healing on disease.

Quantitative data showed perceived significant improvements in ‘concerns/problems’ for which clients wanted help (p<0.01), well-being (p<0.01) and anxiety/depression (p<0.05) over the course of healing. Significant effects were not seen in all areas of quality of life. Qualitative analysis showed clients mainly sought help for psychological and emotional concerns and reported only beneficial effects of healing. Clients attributed many of the quantitative improvements to healing itself. Despite some concerns, healers and clients engaged fully with the research process, and were enthusiastic about the importance of research into healing.

Our study suggests that, while there are some confounding issues and study limitations to address, clients and healers perceive healing to have a range of benefits, particularly in terms of coping with cancer, and regard it as a useful approach that can be applied in a community setting alongside conventional medicine.     

Magnetic resonance imaging of the pancreas at 3.0 tesla: qualitative and quantitative comparison with 1.5 tesla

OBJECTIVES: We sought to perform a preliminary comparison of signal-to-noise ratio (SNR) and image quality for magnetic resonance imaging (MRI) of the pancreas at 1.5 and 3 T. MATERIALS AND METHODS: Two imaging cohorts were studied using a T2-weighted, single-shot fast spin-echo pulse sequence and a T1-weighted, fat-suppressed 3D gradient-echo pulse sequence. In the first cohort, 4 subjects were imaged using identical imaging parameters before and after contrast administration at 1.5 and 3.0 T. The SNR was quantified for the pancreas as well as for the liver, spleen, and muscle. In a second cohort of 12 subjects in whom the receiver bandwidth was adjusted for field strength, SNR measurements and qualitative rankings of image quality were performed. RESULTS: In the study cohort using identical imaging parameters at both magnetic field strengths, the mean (SD) ratios of SNR at 3.0 to 1.5 T of the single-shot fast spin-echo images for the pancreas, liver, spleen, and muscle were 1.63 (0.39), 1.82 (0.39), 1.45 (0.18), 2.01 (0.16), respectively. For the precontrast fat-suppressed 3D gradient-echo sequence, the corresponding ratios were 1.28 (0.29), 1.26 (0.30), 1.16 (0.27), and 1.76 (0.45), respectively; for the arterial phase, the corresponding ratios were 2.02 (0.28), 1.60 (0.42), 1.47 (0.26), and 1.94 (0.32), respectively; and for the delayed postcontrast phase, the corresponding ratios were 1.63 (0.51), 2.01 (0.25), 1.66 (0.06), and 2.31 (0.47), respectively. The SNR benefit of 3.0 T was significantly greater on contrast-enhanced as compared with noncontrast T1-weighted 3D gradient-echo images. In the second study cohort, SNR was superior at 3.0 T, although the use of a reduced readout bandwidth at 1.5 T substantially diminished the advantage of the higher field system. With qualitative comparison of images obtained at the 2 magnetic field strengths, the fat-suppressed 3D gradient-echo images obtained at 3.0 T were preferred, whereas the single shot fast spin-echo images obtained at 1.5 T were preferred because of better signal homogeneity. CONCLUSIONS: Our results in a small cohort of volunteers and patients demonstrate a marked improvement in SNR at 3.0 T compared with 1.5 T (by a factor of 2 in some cases) when identical imaging parameters were used. The SNR advantage at 3.0 T is diminished but persists when the receiver bandwidth is adjusted for magnetic field strength. The results suggest that 3.0 T may offer promise for improved body MRI, although further technical development to optimize SNR and improve signal homogeneity will be needed before its full potential can be achieved.     

Leukocytosis and adverse hospital outcomes after acute myocardial infarction

An elevated white blood cell (WBC) count at the time of hospital presentation is associated with increased mortality after acute myocardial infarction (AMI). The association between WBC count and the development of clinically significant complications of AMI and death during hospitalization for AMI is, however, less clear. The objectives of this observational study were to examine the association between baseline WBC count, the development of heart failure, cardiogenic shock, and death during hospitalization for AMI from a more generalizable community-wide perspective. The study sample consisted of adult residents of all ages from the Worcester, Massachusetts, metropolitan area (1990 census estimate 437,000) hospitalized with confirmed AMI at all greater Worcester medical centers. The study population consisted of 3,796 men and 2,734 women of all ages hospitalized with validated AMI, in 12 annual periods between 1986 and 1999, aggregated into quintiles based on WBC count obtained at the time of hospital admission. In multivariable-adjusted regression analyses controlling for potentially confounding demographic and clinical factors, patients in the uppermost quintiles of WBC count were at increased risk for heart failure (odds ratio [OR] 2.77, 95% confidence interval [CI] 2.33 to 3.31), cardiogenic shock (OR 2.82, 95% CI 2.05 to 3.87), and hospital death (OR 2.14, 95% CI 1.66 to 2.76). The results of our large observational study suggest that the peripheral total leukocyte count is strongly associated with the development of heart failure, cardiogenic shock, and death during hospitalization for AMI. These findings suggest that the WBC count should be considered an important prognostic factor associated with adverse hospital outcomes in patients with AMI.     

A minimally disruptive method for measuring water potential in planta using hydrogel nanoreporters

Leaf water potential is a critical indicator of plant water status, integrating soil moisture status, plant physiology, and environmental conditions. There are few tools for measuring plant water status (water potential) in situ, presenting a critical barrier for developing appropriate phenotyping (measurement) methods for crop development and modeling efforts aimed at understanding water transport in plants. Here, we present the development of an in situ, minimally disruptive hydrogel nanoreporter (AquaDust) for measuring leaf water potential. The gel matrix responds to changes in water potential in its local environment by swelling; the distance between covalently linked dyes changes with the reconfiguration of the polymer, leading to changes in the emission spectrum via Förster Resonance Energy Transfer (FRET). Upon infiltration into leaves, the nanoparticles localize within the apoplastic space in the mesophyll; they do not enter the cytoplasm or the xylem. We characterize the physical basis for AquaDust’s response and demonstrate its function in intact maize (Zea mays L.) leaves as a reporter of leaf water potential. We use AquaDust to measure gradients of water potential along intact, actively transpiring leaves as a function of water status; the localized nature of the reporters allows us to define a hydraulic model that distinguishes resistances inside and outside the xylem. We also present field measurements with AquaDust through a full diurnal cycle to confirm the robustness of the technique and of our model. We conclude that AquaDust offers potential opportunities for high-throughput field measurements and spatially resolved studies of water relations within plant tissues.

Plant life depends on water availability. In managing this demand, irrigated agriculture accounts for 70% of all human water use (1). Physiologically, the process of transpiration (E) dominates this demand for water (Fig. 1A): Solar thermal radiation and the unsaturated relative humidity in the atmosphere drive evaporation from the wet internal surfaces of leaves; this water loss pulls water up through the plant’s vascular tissue (xylem) and out of the soil. This flow occurs along a gradient in the chemical potential of water, or water potential, ψ [MPa] (2). Studies of water relations and stress physiology over the past decades have found that values of ψ along the path of E (the soil–plant–atmosphere continuum [SPAC]) correlate with plant growth, crop yield and quality, susceptibility to disease, and the balance between water loss due to E and the uptake and assimilation of carbon dioxide (water-use efficiency) (3–5).Open in a separate windowFig. 1.AquaDust as an in situ reporter of water potential (ψ). (A) Schematic representation of a maize plant undergoing transpiration (E) in a dynamic environment driven by solar thermal radiation (Qrad) and photosynthetically active radiation (PAR), wind speed (u), temperature (T), vapor pressure deficit (VPD), and soil water potential (ψsoil). Water flows through the plant (blue arrows) along a gradient in water potential (∇→ψ). Zones on the leaves infiltrated with AquaDust serve as reporters of the local leaf water potential, ψleaf, via a short (∼30 s), minimally invasive measurement of FRET efficiency (ζ) with a leaf clamp. (B) Schematic representations of infiltration of a suspension of AquaDust and of the distribution of AquaDust within the cross-section of a leaf. AquaDust passes through the stomata and localizes in the apoplastic spaces within the mesophyll; the particles are excluded from symplastic spaces and the vascular bundle. (C) Schematic diagrams showing mechanism of AquaDust response: The swollen, “wet” state when water potential in its local environment, ψenv=0 (i.e., no stress condition), results in low FRET between donor (green circles) and acceptor (yellow circles) dye (Upper); and the shrunken, “dry” state when ψenv<0 (i.e., stressed condition) results in high FRET between fluorophores, thereby altering the emission spectra (Lower). (D) Fluorescent dyes were chosen to minimize reabsorption of AquaDust emission from chlorophyll; comparison of representative fluorescent emission from AquaDust (donor peak at 520 nm and acceptor peak at 580 nm) with the absorption spectra of chlorophyll and autofluorescence of maize leaf.Due to the recognized importance of water potential in controlling plant function, plant scientists have spent considerable effort devising accurate and reliable methods to measure water potential of the soil, stem, and leaf (6). Of these, plant water potentials, and particularly leaf water potential (ψleaf), represent valuable indicators of plant water status because they integrate both environmental conditions (e.g., soil water availability and evaporative demand) and plant physiological processes (e.g., root water uptake, xylem transport, and stomatal regulation) (7, 8). To date, techniques to measure ψleaf remain either slow, destructive, or indirect. The current tools (e.g., Scholander pressure chamber, psychrometer, and pressure probe) involve disruption of the tissue, the microenvironment, or both (9–11). For example, the widely used pressure chamber requires excision of leaves or stems for the measurement of ψleaf. Other techniques, such as stem and leaf psychrometry, require intimate contact with the tissue, and accurate and repeatable measurements are difficult to obtain (9, 12). These limitations have hindered the study of spatiotemporal water-potential gradients along the SPAC and the development of high-throughput strategies to phenotype based on tissue water potential (13). Additionally, current methods for measuring ψleaf provide averages over tissues in the leaf. This characteristic makes the dissection of water relations on subleaf scales challenging, such that important questions remain, for example, about the partitioning of hydraulic resistances within leaves between the xylem and mesophyll (14–16).These outstanding challenges in the measurement of water status in planta motivated us to develop the measurement strategy presented here, AquaDust, with the following characteristics: 1) Minimally disruptive: Compatible with simple, rapid measurements on intact leaves. Fig. 1A presents our approach, in which AquaDust reporters infiltrated into the mesophyll of the leaf provide an externally accessible optical signal that correlates with the local water potential. 2) Localized: allowing for access to the values of water potential at a well-defined location along the path of transpiration in the leaf tissue. Fig. 1B shows a schematic representation of AquaDust particles localized in the apoplastic volume within the mesophyll, at the end of the hydraulic path for liquid water within the plant. 3) Sensitive and specific: capable of resolving water potentials across the physiologically relevant range (∼−3<ψ<0 MPa) and with minimal sensitivity to other physical (e.g., temperature) and chemical (e.g., pH) variables. Fig. 1C presents a schematic representation of an AquaDust particle formed of hydrogel, a highly tunable material that undergoes a structural response to changes in local water potential (swollen when wet; collapsed when dry). We couple the swelling behavior of the particle to an optical signal via the incorporation of fluorescence dyes (green and yellow circles in Fig. 1C) that undergo variable Förster Resonance Energy Transfer (FRET) as a function of spatial separation. Fig. 1D presents typical AquaDust spectra at high (wet; green curve) and low (dry; yellow curve) water potentials. A change in water potential leads to a change in the relative intensity of the two peaks in the AquaDust spectrum, such that the relative FRET efficiency, ζ=f(ID,IA), can serve as a measure of water potential. 4) Inert: nondisruptive of the physiological properties of the leaf (e.g., photosynthetic capacity, transpiration rate, etc.).In this paper, we present the development, characterization, and application of AquaDust. We show that AquaDust provides a robust, reproducible response of its fluorescence spectra to changes in leaf water potential in situ and across the usual physiological range. We apply our approach to quantify the spatial gradients of water potential along individual leaves undergoing active transpiration and across a range of soil water potentials. With these measurements, we show that the localization of AquaDust in the mesophyll allows us to quantify the importance of hydraulic resistances outside the xylem. We further use AquaDust to measure the diurnal dynamics of ψleaf under field conditions, with repeated measurements on individual, intact leaves. These measurements demonstrate the field-readiness of our techniques and validate the leaf hydraulic model we have developed. We conclude that AquaDust offers a powerful basis for tracking, spatially and temporally, water potential in planta to study the mechanisms by which it couples to both biological and physical processes to define plant function.     

Crystal chemistry and single-phase synthesis of Gd3+ substituted Co–Zn ferrite nanoparticles for enhanced magnetic properties

Rare earth (RE) ions are known to improve the magnetic interactions in spinel ferrites if they are accommodated in the lattice, whereas the formation of a secondary phase leads to the degradation of the magnetic properties of materials. Therefore, it is necessary to solubilize the RE ions in a spinel lattice to get the most benefit. In this context, this work describes the synthesis of Co–Zn ferrite nanoparticles and the Gd³⁺ doping effect on the tuning of their magnetic properties. The modified sol–gel synthesis approach offered a facile way to synthesize ferrite nanoparticles using water as the solvent. X-ray diffraction with Rietveld refinement confirmed that both pure Co–Zn ferrite and Gd³⁺ substituted Co–Zn ferrite maintained single-phase cubic spinel structures. Energy dispersive spectroscopy was used to determine the elemental compositions of the nanoparticles. Field and temperature dependent magnetic characteristics were measured by employing a vibration sample magnetometer in field cooled (FC)/zero field cooled (ZFC) modes. Magnetic interactions were also determined by Mössbauer spectroscopy. The saturation magnetization and coercivity of Co–Zn ferrite were improved with the Gd³⁺ substitution due to the Gd³⁺ (4f⁷)–Fe³⁺ (3d⁵) interactions. The increase in magnetization and coercivity makes these Gd³⁺ substituted materials applicable for use in magnetic recording media and permanent magnets.

Rare earth (RE) ions are known to improve the magnetic interactions in spinel ferrites if they are accommodated in the lattice, whereas the formation of a secondary phase leads to the degradation of the magnetic properties of materials.     

Detection of recombinant human erythropoietin abuse in athletes utilizing markers of altered erythropoiesis

BACKGROUND AND OBJECTIVES: The detection of recombinant human erythropoietin (r-HuEPO) abuse by athletes remains problematic. The main aim of this study was to demonstrate that the five indirect markers of altered erythropoiesis identified in our earlier work were reliable evidence of current or recently discontinued r-HuEPO use. A subsidiary aim was to refine weightings of the five markers in the initial model using a much larger data set than in the pilot study. A final aim was to verify that the hematologic response to r-HuEPO did not differ between Caucasian and Asiatic subjects. DESIGN AND METHODS: Recreational athletes resident in Sydney, Australia (Sydney, n = 49; 16 women, 33 men) or Beijing, China (Beijing, n=24; 12 women, 12 men) were randomly assigned to r-HuEPO or placebo groups prior to a 25 day administration phase. Injections of r-HuEPO (or saline) were administered double-blind at a dose of 50 IU/kg three times per week, with oral iron (105 mg) or placebo supplements taken daily by all subjects. Blood profiles were monitored during and for 4 weeks after drug administration for hematocrit (Hct), reticulocyte hematocrit (RetHct), percent macrocytes (%Macro), serum erythropoietin (EPO) and soluble transferrin receptor (sTfr), since we had previously shown that these five variables were indicative of r-HuEPO use. RESULTS. The changes in Hct, RetHct, %Macro, EPO and sTfr in the Sydney trial were qualitatively very similar to the changes noted in our previous administration trial involving recreational athletes of similar genetic origin. Statistical models developed from Fisher's discriminant analysis were able to categorize the user and placebo groups correctly. The same hematologic response was demonstrated in Beijing athletes also administered r-HuEPO. INTERPRETATION AND CONCLUSIONS: This paper confirms that r-HuEPO administration causes a predictable and reproducible hematologic response. These markers are disturbed both during and for several weeks following r-HuEPO administration. This work establishes an indirect blood test which offers a useful means of detecting and deterring r-HuEPO abuse. Ethnicity did not influence the markers identified as being able to detect athletes who abuse r-HuEPO.     

Inhibited apoptosis and drug resistance in acute myeloid leukaemia

Despite extensive investigation into mechanisms of drug resistance in acute myeloid leukaemia (AML), the aetiology of therapeutic resistance is unclear. We found that five leukaemia cell lines (K562, HL-60, CEM, CEM induced to overexpress bcl-2, and REH) displayed parallel sensitivity to four antileukaemia drugs with different mechanisms of action, with K562 generally being the least sensitive and REH being the most sensitive. The amount of spontaneous apoptosis in the cell lines after serum-free culture paralleled their drug sensitivity: K562 cells displayed the least apoptosis at 24 h (2.50 ± 0.24%) and REH the most (24.47 ± 8.22%). The extent of spontaneous apoptosis of leukaemic blasts from 39 patients with newly diagnosed de novo AML also correlated with the success of the intensive, infusional cytarabine-based induction therapy. There was a median of 19.5% (range 3.6–64%) apoptotic AML cells after 24 h of serum-free culture in patients who entered a complete remission compared with 4.2% (1.8–7.0%) apoptotic AML cells in patients who did not achieve a complete remission ( P  = 0.0007). Thus, inhibited apoptosis was associated with both in vitro and in vivo pan-resistance to antileukaemic chemotherapy. The cause of inhibited apoptosis in AML is probably a function of interactions among multiple signals that influence apoptosis. Assessment of spontaneous apoptosis may serve as an important prognostic factor for AML.     

Prognosis following interventional therapy for acute myocardial infarction: utility of dipyridamole thallium scintigraphy.

A variety of methods for risk stratification after acute myocardial infarction have been successfully employed, however, little attention has been focused on patients who have received reperfusion therapy. The present report examines the utility of dipyridamole thallium scintigraphy in the prediction of late cardiac death or recurrent myocardial infarction in patients who have received thrombolytic therapy. Prospectively, 71 patients who presented with myocardial infarction and were treated with recombinant tissue plasminogen activator (and frequently percutaneous transluminal coronary angioplasty) were enrolled in the study. The primary end points during the follow-up period of nearly 2 years were recurrent infarction or death, which occurred in 10 patients. Although cardiac events were significantly related to either the performance of late myocardial revascularization or the presence of a residual coronary artery stenosis at discharge, no scintigraphic variable was found to be predictive of myocardial infarction or death. Thus, this report is the first to suggest limitation of scintigraphic techniques with regard to prognostic value in myocardial infarction survivors treated with reperfusion techniques. This selected population may have physiologic differences as compared with post-infarction studies performed before the advent of thrombolytic agents. Caution is therefore advised in extrapolating results of earlier reports to the ever increasing percentage of patients receiving recannalization therapy.