The clinical application of macromolecular Gd(III) complexes as MRI contrast agents is impeded by their slow excretion and potential toxicity due to the release of Gd(III) ions caused by the ...metabolism of the agents. A polymer Gd(III) chelate conjugate with a cleavable spacer has been designed to solve this problem. Poly(l-glutamic acid)−cystamine−Gd(III)-DOTA was prepared by the conjugation of DOTA to PGA (MW = 50 000) via cystamine, a cleavable disulfide spacer, followed by the complexation with GdCl3. A Gd(III) DOTA chelate derivative was readily released from the polymer conjugate in the incubation with cysteine, an endogenous plasma thiol. The conjugate produced significant MRI blood pool contrast enhancement in nude mice bearing OVCAR-3 human ovarian carcinoma xenographs. Less significant contrast enhancement was observed for a small molecular contrast agent, Gd(DTPA-BMA). The pharmacokinetic MRI study showed that the Gd(III) chelate from the conjugate accumulated in the urinary bladder in a similar kinetic pattern to Gd(DTPA-BMA), suggesting that the chelate was released by the endogenous thiols and excreted through renal filtration. The preliminary results suggest that this novel design has a great potential to solve the safety problem of macromolecular MRI contrast agents.
To meet international greenhouse gas reporting obligations, New Zealand must report on carbon stocks in forests established after 1989 (post-1989 forest). Although predominately comprised of planted ...forest, post-1989 forest also contains a component of natural vegetation amounting to less than 10% by area. New Zealand undertook a national inventory of this natural stratum of post-1989 forest to provide estimates of carbon stocks and stock change in woody species over the first commitment period (2008–2012) of the Kyoto Protocol. Plots were installed on a 4-km grid, and the basal diameters and heights of trees and shrubs were measured for the first time from November 2012, to March 2013. Carbon stocks in 2012 were calculated using allometric functions developed from biomass samples from each site. Basal disc samples provided data on diameter increment and shrub and tree age annually from 1990 to 2012. These were used to predict carbon stocks per ha for individual plots in 2008 and to provide annual predictions by pool back to 1990. Carbon stocks summed across live and dead biomass pools (excluding soil) averaged 3.04, 16.70 and 28.73 t C/ha in 1990, 2008 and 2012, respectively. The disposition by pool was 2.25, 12.54 and 21.84 t C/ha in aboveground biomass, 0.56, 3.13 and 5.46 t C/ha in belowground biomass (using a root/shoot ratio of 0.25), 0.03, 0.17 and 0.23 t C/ha in deadwood, and 0.18, 0.86 and 1.21 t C/ha in litter in 1990, 2008 and 2012, respectively. In 1990, the woody biomass stock estimate per plot ranged from zero to 40 t C/ha and averaged 3.04 t C/ha across all plots. The methodology used to predict annual carbon stocks required an assumption concerning stem annual mortality. Sensitivity analysis suggested that varying this assumption had only a minor impact on predicted carbon stocks and changes. Plant age varied markedly within and between the natural forest plots, and therefore, the mean age of woody vegetation at each site was obtained by setting a threshold woody biomass carbon stock that needed to be achieved, and vegetation age was calculated as years since the threshold was achieved. This threshold approach facilitated the development of a yield table for predicting carbon (t/ha) as a function of vegetation mean age.
The Theory of Critical Distances groups together a number of approaches postulating that, in cracked/notched materials subjected to static loading, breakage takes place as soon as a critical ...length-dependent effective stress exceeds the material tensile strength. The characteristic length used by the Theory of Critical Distances is a material property that can directly be estimated from the ultimate tensile strength and the plane strain fracture toughness. In the present investigation, based on a large number of bespoke experimental results, it is demonstrated that the Theory of Critical Distances is successful also in quantifying the detrimental effect of cracks and manufacturing defects in 3D-printed concrete subjected to Mode I static loading.
Background and purpose:
The TASK subfamily of two pore domain potassium channels (K2P) encodes for leak K currents, contributing to the resting membrane potential of many neurons and regulating their ...excitability. TASK1 and TASK3 channels are regulated by a number of pharmacological and physiological mediators including cannabinoids such as methanandamide. In this study, we investigate how methanandamide blocks these channels.
Experimental approach:
Currents through wild type and mutated TASK1 and TASK3 channels expressed in modified HEK‐293 cells were measured using whole‐cell electrophysiological recordings in the presence and absence of methanandamide.
Key results:
Methanandamide (3 μM) produced substantial block of hTASK1, hTASK3 and mTASK3 channels but was most potent at blocking hTASK3 channels. Block of these channels was irreversible unless cells were washed with buffer containing bovine serum albumin. Mutation of the distal six amino acids of TASK1 did not alter methanandamide inhibition, whilst C terminal truncation of TASK3 channels caused a small but significant reduction of inhibition. However, deletion of six amino acids (VLRFLT) at the interface between the final transmembrane domain and cytoplasmic C terminus of TASK3 channels gave functional currents that were no longer inhibited by methanandamide or by activation of GPCRs.
Conclusions and implications:
Methanandamide potently blocked TASK3 and TASK1 channels and both methanandamide and G protein‐mediated inhibition converged on the same intracellular gating pathway. Physiologically, methanandamide block of TASK1 and TASK3 channels may underpin a number of CNS effects of cannabinoids that are not mediated through activation of CB1 or CB2 receptors.
British Journal of Pharmacology (2007) 152, 778–786. doi:10.1038/sj.bjp.0707436; published online 10 September 2007
Background. A decline of hepatitis C virus (HCV) antibody titers (anti-HCV), ultimately resulting in seroreversion, has been reported following clearance of viremia in both acute and chronic HCV ...infection. However, frequency of seroreversion remains unknown in human immunodeficiency virus (HIV)/HCV-coinfected patients. We describe anti-HCV dynamics among HIV-infected men who have sex with men (MSM) following acute HCV infection and reinfection. Methods. Primary acute HCV infection was assumed when a subject was anti-HCV negative prior to the first positive HCV RNA test. Anti-HCV was measured at least annually in 63 HIV-infected MSM, with a median follow-up of 4.0 years (interquartile range IQR, 2.5–5.7 years). Time from HCV infection to seroconversion, and from seroconversion to seroreversion, was estimated using the Kaplan-Meier method. Longitudinal anti-HCV patterns were studied using a random-effects model to adjust for repeated measures. Results. Median time from HCV infection to seroconversion was 74 days (IQR, 47–125 days). Subjects who cleared HCV RNA (n = 36) showed a significant decrease in anti-HCV levels (P<.001). Among 31 subjects with sustained virologic response (SVR), anti-HCV became undetectable during follow-up in 8; cumulative incidence of seroreversion within 3 years after seroconversion was 37% (95% confidence interval, 18%–66%). Eighteen subjects became reinfected during follow-up; this coincided with a subsequent increase in anti-HCV reactivity. Conclusions. A decline of anti-HCV reactivity was associated with HCV RNA clearance. Seroreversion was very common following SVR. Upon reinfection, anti-HCV levels increased again. Monitoring anti-HCV levels might therefore be an effective alternative for diagnosis of HCV reinfection.
The much-praised Cultural Quarters returns in a revised edition, offering new case studies and new chapters on the economics of cultural quarters and the importance of historical buildings. This ...definitive text provides a conceptual context for cultural quarters through a detailed discussion of urban design and planning.
Cardiovascular disease is the leading cause of morbidity and mortality in the world. Atherosclerotic plaques, consisting of lipid-laden macrophages and calcification, develop in the coronary ...arteries, aortic valve, aorta, and peripheral conduit arteries and are the hallmark of cardiovascular disease. In humans, imaging with computed tomography allows for the quantification of vascular calcification; the presence of vascular calcification is a strong predictor of future cardiovascular events. Development of novel therapies in cardiovascular disease relies critically on improving our understanding of the underlying molecular mechanisms of atherosclerosis. Advancing our knowledge of atherosclerotic mechanisms relies on murine and cell-based models. Here, a method for imaging aortic calcification and macrophage infiltration using two spectrally distinct near-infrared fluorescent imaging probes is detailed. Near-infrared fluorescent imaging allows for the ex vivo quantification of calcification and macrophage accumulation in the entire aorta and can be used to further our understanding of the mechanistic relationship between inflammation and calcification in atherosclerosis. Additionally, a method for isolating and culturing animal aortic vascular smooth muscle cells and a protocol for inducing calcification in cultured smooth muscle cells from either murine aortas or from human coronary arteries is described. This in vitro method of modeling vascular calcification can be used to identify and characterize the signaling pathways likely important for the development of vascular disease, in the hopes of discovering novel targets for therapy.
Influenza hemagglutinin (HA) undergoes a conformational change that is required for viral entry. The rearrangement includes exposure of the fusion peptide, a hydrophobic segment buried in the trimer ...interface of the native protein. Since fusion peptide release triggers the membrane fusion event crucial for viral replication, inhibition of fusion peptide exposure should prevent infection. We reasoned that small molecules that bind to HA and stabilize its nonfusogenic conformation would block viral activity. A computer-assisted method was used to select putative HA ligands. One of the selected compounds, 4A,5,8,8A-tetrahydro-5,8-methano-1,4-naphthoquinone, prevented the conversion of X31 HA to a conformation recognized by alpha-fusion peptide antisera. Several derivatives of this compound, including both benzoquinones and hydroquinones, also showed inhibition. The most effective compounds tested have IC50S between 1 and 20 microM. Representative compounds also inhibited virus-induced syncytia formation, HA-mediated hemolysis, and viral infectivity in vitro. The inhibitors are attractive leads for the development of antiviral drugs and can serve as probes of the mechanism of the conformational change of HA.
A series of model steels and commercial alloys from the IAEA Phase 3 irradiation programme was irradiated to a dose of 14 mdpa (∼ 9 × 10
22 n m
−2 (
E > 1 MeV)) at 290°C. The steels were examined ...using small angle neutron scattering and a range of transmission electron microscope techniques. Measured yield stress and hardness changes were interpreted in terms of the irradiation-induced microstructural developments. The irradiation of Cu-containing steels produced Cu-rich precipitates of ∼ 2 nm diameter which were alloyed with Mn and Ni. The Ni content of the precipitates increased markedly for bulk Ni contents > 0.7 wt%. The volume fractions of precipitate increased with increasing bulk Cu content and, also, with increasing bulk Ni content > 0.7 wt%. The precipitate-induced strengthening and hardening varied according to the square root of the volume fraction of precipitate. Yield stress changes in low-Cu steels were consistent with data from the IAEA Phase 2 irrradiation programme.
Results are presented from controlled field tests of two methods for detecting and diagnosing faults in HVAC equipment. The tests were conducted in a unique research building that featured two ...air-handling units serving matched sets of unoccupied rooms with adjustable internal loads. Tests were also conducted on a third air handler in the same building, serving areas used for instruction and by building staff. One of the two fault detection and diagnosis (FDD) methods used first-principles-based models of system components. The data used by this approach were obtained from sensors typically installed for control purposes. The second method was based on semiempirical correlations of submetered electrical power with flow rates or process control signals. Faults were introduced into the air-mixing, filter-coil, and fan sections of each of the three air-handling units. In the matched air-handling units, faults were implemented over three blind test periods (summer, winter, and spring operating conditions). In each test period, the precise timing of the implementation of the fault conditions was unknown to the researchers. The faults were, however, selected from an agreed set of conditions and magnitudes, established for each season. This was necessary to ensure that at least some magnitudes of the faults could be detected by the FDD methods during the limited test period. Six faults were used for a single summer test period involving the third air-handling unit. These fault conditions were completely unknown to the researchers and the test period was truly blind. The two FDD methods were evaluated on the basis of their sensitivity, robustness, the number of sensors required, and ease of implementation. Both methods detected nearly all of the faults in the two matched air-handling units but fewer of the unknown faults in the third air-handling unit. Fault diagnosis was more difficult than detection. The first-principles-based method misdiagnosed several faults. The electrical power correlation method demonstrated greater success in diagnosis, although the limited number of faults addressed in the tests contributed to this success. The first-principles-based models require a larger number of sensors than the electrical power correlation models, although the latter method requires power meters that are not typically installed. The first-principles-based models require training data for each subsystem model to tune the respective parameters so that the model predictions more precisely represent the target system. This is obtained by an open-loop test procedure. The electrical power correlation method uses polynomial models generated from data collected from 'normal' system operation, under closed-loop control. Both methods were found to require further work in three principal areas: to reduce the number of parameters to be identified; to assess the impact of less expensive or fewer sensors; and to further automate their implementation. The first-principles-based models also require further work to improve the robustness of predictions. The full text of this paper can be found in the International Journal of Heating, Ventilating, Air-Conditioning and Refrigerating Research, Volume 8, Number 1, January 2002, pp. 41-71.
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