BCR-ABL1 genomic DNA PCR response kinetics during first-line imatinib treatment of chronic myeloid leukemia

Accurate quantification of minimal residual disease during treatment of chronic myeloid leukaemia guides clinical decisions. The conventional minimal residual disease method, RQ-PCR for BCR-ABL1 mRNA, reflects a composite of the number of circulating leukemic cells and the BCR-ABL1 transcripts per cell. BCR-ABL1 genomic DNA only reflects leukemic cell number. We used both methods in parallel to determine the relative contribution of the leukemic cell number to molecular response. BCR-ABL1 DNA PCR and RQ-PCR were monitored up to 24 months in 516 paired samples from 59 newly-diagnosed patients treated with first-line imatinib in the TIDEL-II study. In the first 3 months of treatment BCR-ABL1 mRNA values declined more rapidly than DNA. By 6 months the two measures aligned closely. The expression of BCR-ABL1 mRNA was normalized to cell number to generate an expression ratio. The expression of e13a2 BCR-ABL1 was lower than that of e14a2 transcripts at multiple time points during treatment. BCR-ABL1 DNA was quantifiable in 48% of samples with undetectable BCR-ABL1 mRNA, resulting in minimal residual disease being quantifiable for an additional 5-18 months (median 12 months). These parallel studies show for the first time that the rapid decline in BCR-ABL1 mRNA over the first 3 months of treatment is due to a reduction in both cell number and transcript level per cell, whereas beyond 3 months falling levels of BCR-ABL1 mRNA are predominantly due to depletion of leukaemic cells.Ilaria S. Pagani, Phuong Dang, Ivar O. Kommers, Jarrad M. Goyne, Mario Nicola, Verity A. Saunders, Jodi Braley, Deborah L. White, David T. Yeung, Susan Branford, Timothy P. Hughes, and David M. Ros

Loss of Grem1-lineage chondrogenic progenitor cells causes osteoarthritis

Published online: 31 October 2023Osteoarthritis (OA) is characterised by an irreversible degeneration of articular cartilage. Here we show that the BMP-antagonist Gremlin 1 (Grem1) marks a bipotent chondrogenic and osteogenic progenitor cell population within the articular surface. Notably, these progenitors are depleted by injury-induced OA and increasing age. OA is also caused by ablation of Grem1 cells in mice. Transcriptomic and functional analysis in mice found that articular surface Grem1-lineage cells are dependent on Foxo1 and ablation of Foxo1 in Grem1-lineage cells caused OA. FGFR3 signalling was confirmed as a promising therapeutic pathway by administration of pathway activator, FGF18, resulting in Grem1-lineage chondrocyte progenitor cell proliferation, increased cartilage thickness and reduced OA. These findings suggest that OA, in part, is caused by mechanical, developmental or age-related attrition of Grem1 expressing articular cartilage progenitor cells. These cells, and the FGFR3 signalling pathway that sustains them, may be effective future targets for biological management of OA.Jia Q. Ng, Toghrul H. Jafarov, Christopher B. Little, Tongtong Wang, Abdullah M. Ali, YanMa, Georgette A. Radford, Laura Vrbanac, Mari Ichinose, Samuel Whittle, David J. Hunter, Tamsin R. M. Lannagan, Nobumi Suzuki, JarradM. Goyne, Hiroki Kobayashi, Timothy C. Wang, David R. Haynes, Danijela Menicanin, Stan Gronthos, Daniel L. Worthley, Susan L. Woods and Siddhartha Mukherje

Soil water extraction dynamics of dryland cotton in various row configurations

The soil water extraction dynamics of dryland cotton grown in various row configurations on some major Queensland cotton soils are being quantified. The data will assist in the further development of a cotton simulation model "CERCOT". This model is to be used to determine the outcome of various management scenarios relating row configuration, planting time and plant available water supply. This paper presents some results obtained on one soil type during the 1997/98 season

Radiation interception, radiation use efficiency and growth of barley cultivars

Dry matter production and utilization of photosynthetically active radiation (PAR) was studied for barley (Hordeurn vulgare L.) in the field at Hermitage Research Station, Qld. In 1990, four cultivars (Gilbert, Tallon, Grimmett, Skiff) were sown at three times and grown with non-limiting soil moisture. In 1991, soil moisture limitations were imposed on one sowing of the cultivar Grimmett. The radiation extinction coefficient (k) was 0.41±0.02 and did not vary with cultivar, time of sowing or soil moisture availability. Radiation use efficiency (RUE) (based on absorbed PAR and above-ground dry matter) did not change with time of sowing but did vary between cultivars. RUE was highest for Gilbert (2.90±0.10 g MJ-1), while the other three cultivars averaged 2.60±0.04 g MJ-l. RUE of Grimmett was significantly lower in 1991 (1.48±0.07 g MJ-1) than in 1990 (2.60±0.07g MJ-1), but soil moisture differences in 1991 did not significantly affect RUE. Several factors with possible links with RUE were examined and discussed. Of the variables examined those which showed the strongest relationships with RUE were average daily vapour pressure deficit and average daily minimum temperature

Development and use of a barley crop simulation model to evaluate production management strategies in north-eastern Australia

A study was undertaken to identify improved management strategies for barley (Hordeum vulgare L.), particularly in relation to time of planting, location, and frost risk in the variable climate of north-eastern Australia. To achieve this objective, a crop growth simulation model (QBAR) was constructed to integrate the understanding, gained from field experiments, of the dynamics of crop growth as influenced by soil moisture and environmental variables. QBAR simulates the growth and yield potential of barley grown under optimal nutrient supply, in the absence of pests, diseases, and weeds. Genotypic variables have been determined for 4 cultivars commonly grown in the northern cereal production areas. Simulations were conducted using long- term weather data to generate the probabilistic yield outcome of cv. Grimmer for a range of times of planting at 10 locations in the north-eastern Australian grain belt. The study indicated that the common planting times used by growers could be too late under certain circumstances to gain full yield potential. Further applications of QBAR to generating information suitable for crop management decision support packages and crop yield forecasting are discussed

A novel ruthenium-based molecular sensor to detect endothelial nitric oxide

Nitric oxide (NO) is a key regulator of endothelial cell and vascular function. The direct measurement of NO is challenging due to its short half-life, and as such surrogate measurements are typically used to approximate its relative concentrations. Here we demonstrate that ruthenium-based [Ru(bpy)2(dabpy)]2+ is a potent sensor for NO in its irreversible, NO-bound active form, [Ru(bpy)2(T-bpy)]2+. Using spectrophotometry we established the sensor's ability to detect and measure soluble NO in a concentration-dependent manner in cell-free media. Endothelial cells cultured with acetylcholine or hydrogen peroxide to induce endogenous NO production showed modest increases of 7.3 ± 7.1% and 36.3 ± 25.0% respectively in fluorescence signal from baseline state, while addition of exogenous NO increased their fluorescence by 5.2-fold. The changes in fluorescence signal were proportionate and comparable against conventional NO assays. Rabbit blood samples immediately exposed to [Ru(bpy)2(dabpy)]2+ displayed 8-fold higher mean fluorescence, relative to blood without sensor. Approximately 14% of the observed signal was NO/NO adduct-specific. Optimal readings were obtained when sensor was added to freshly collected blood, remaining stable during subsequent freeze-thaw cycles. Clinical studies are now required to test the utility of [Ru(bpy)2(dabpy)]2+ as a sensor to detect changes in NO from human blood samples in cardiovascular health and disease.Achini K. Vidanapathirana, Benjamin J. Pullen, Run Zhang, MyNgan Duong, Jarrad M. Goyne, Xiaozhou Zhang, Claudine S. Bonder, Andrew D. Abell, Christina A. Bursill, Stephen J. Nicholls, Peter J. Psalti

Biological Sensing of Nitric Oxide in Macrophages and Atherosclerosis Using a Ruthenium-Based Sensor

Macrophage-derived nitric oxide (NO) plays a critical role in atherosclerosis and presents as a potential biomarker. We assessed the uptake, distribution, and NO detection capacity of an irreversible, ruthenium-based, fluorescent NO sensor (Ru-NO) in macrophages, plasma, and atherosclerotic plaques. In vitro, incubation of Ru-NO with human THP1 monocytes and THP1-PMA macrophages caused robust uptake, detected by Ru-NO fluorescence using mass-cytometry, confocal microscopy, and flow cytometry. THP1-PMA macrophages had higher Ru-NO uptake (+13%, p < 0.05) than THP1 monocytes with increased Ru-NO fluorescence following lipopolysaccharide stimulation (+14%, p < 0.05). In mice, intraperitoneal infusion of Ru-NO found Ru-NO uptake was greater in peritoneal CD11b+F4/80+ macrophages (+61%, p < 0.01) than CD11b+F4/80− monocytes. Infusion of Ru-NO into Apoe−/− mice fed high-cholesterol diet (HCD) revealed Ru-NO fluorescence co-localised with atherosclerotic plaque macrophages. When Ru-NO was added ex vivo to aortic cell suspensions from Apoe−/− mice, macrophage-specific uptake of Ru-NO was demonstrated. Ru-NO was added ex vivo to tail-vein blood samples collected monthly from Apoe−/− mice on HCD or chow. The plasma Ru-NO fluorescence signal was higher in HCD than chow-fed mice after 12 weeks (37.9%, p < 0.05). Finally, Ru-NO was added to plasma from patients (N = 50) following clinically-indicated angiograms. There was lower Ru-NO fluorescence from plasma from patients with myocardial infarction (−30.7%, p < 0.01) than those with stable coronary atherosclerosis. In conclusion, Ru-NO is internalised by macrophages in vitro, ex vivo, and in vivo, can be detected in atherosclerotic plaques, and generates measurable changes in fluorescence in murine and human plasma. Ru-NO displays promising utility as a sensor of atherosclerosis.Achini K. Vidanapathirana, Jarrad M. Goyne, Anna E. Williamson, Benjamin J. Pullen, Pich Chhay, Lauren Sandeman, Julien Bensalem, Timothy J. Sargeant, Randall Grose, Mark J. Crabtree, Run Zhang, Stephen J. Nicholls, Peter J. Psaltis, and Christina A. Bursil

Supersonic combustion of hydrogen in Mach 2 flows over an axisymmetric model

An external axisymmetric model featuring a cavity flame-holder fuelled by hydrogen was adopted for the study of the supersonic combustion in Mach 2 flows. The self-sustained hydrogen flames ignited by a spark gap igniter were detected using two optical techniques: an intensified camera attachment coupled with a high speed camera with a narrow-band-pass filter centred at 310 nm for detection of OH* chemiluminescence; and high-speed schlieren imaging for flow visualization. The spark ignition was identified on the schlieren images which also enabled visualisation of changes in the flow structure associated with the combustion. Local concentrations of the radiating radical OH* were determined using the inverse Abel transformation based on the ICCD images and calibration against a light source of known radiance. The combustion flow was reconstructed numerically using a CFD Solver, Eilmer4 in an axisymmetric configuration coupled with a detailed H2/O2 oxidation chemistry mechanism and a OH* chemiluminescence sub-scheme. The maximum magnitude of the results for the OH* chemiluminescence simulations demonstrated reasonable agreement with measurements