Effect of changing the fine particle mass of inhaled beclomethasone dipropionate on intrapulmonary deposition and pharmacokinetics

AbstractReformulation of beclomethasone dipropionate (BDP) in the chlorofluorocarbon (CFC)-free propellant hydrofluoroalkane-134a (HFA) gave the opportunity to produce a solution formulation that provides a greater total mass of fine drug particles than the current CFC suspension metered dose inhaler (MDI). The HFA-BDP MDI was studied in three pharmacokinetic trials in asthmatic patients. Serum levels of BDP plus metabolites [total beclomethasone (total BOH) assay] were used to test whether the increased fine particle mass of HFA-BDP would result in improved intrapulmonary deposition and subsequent differences in serum profiles. Serum levels, maximum serum concentrations and area under the serum concentration-time curves of total BOH following both single and multiple doses of HFA-BDP were similar to those obtained with approximately twice the dose of CFC-BDP. The observed lower bioavailability of CFC-BDP compared with HFA-BDP could be explained if most of each inhaled dose from the CFC-BDP MDI was swallowed and absorbed from the gastrointestinal tract, while most of each inhaled dose from the HFA-BDP MDI was absorbed from the lungs. Deposition studies have confirmed this explanation. These results suggest that asthmatic patients can be treated with lower total daily doses of drug from HFA-BDP extrafine aerosol than from CFC-BDP

On Single-Cell Enzyme Assays in Marine Microbial Ecology and Biogeochemistry

Extracellular enzyme activity is a well-established parameter for evaluating microbial biogeochemical roles in marine ecosystems. The presence and activity of extracellular enzymes in seawater provide insights into the quality and quantity of organic matter being processed by the present microorganisms. A key challenge in our understanding of these processes is to decode the extracellular enzyme repertoire and activities of natural communities at the single-cell level. Current measurements are carried out on bulk or size-fractionated samples capturing activities of mixed populations. This approach – even with size-fractionation – cannot be used to trace enzymes back to their producers, nor distinguish the active microbial members, leading to a disconnect between measured activities and the producer cells. By targeting extracellular enzymes and resolving their activities at the single-cell level, we can investigate underlying phenotypic heterogeneity among clonal or closely related organisms, characterize enzyme kinetics under varying environmental conditions, and resolve spatio-temporal distribution of individual enzyme producers within natural communities. In this perspective piece, we discuss state-of-the-art technologies in the fields of microfluidic droplets and functional screening of prokaryotic cells for measuring enzyme activity in marine seawater samples, one cell at a time. We further elaborate on how this single-cell approach can be used to address research questions that cannot be answered with current methods, as pertinent to the enzymatic degradation of organic matter by marine microorganisms

Pachinko Biology: Gambling on Single Cells

While stem cells hold great promise as a means of treating otherwise incurable disease due to their characteristic ability to both self-renew and differentiate, they can also play a role in disease. Chronic myeloid leukemia (CML) arises in CD34+ hematopoietic stem, and is known resistant to the tyrosine kinase inhibitors imatinib and dasatinib. Understanding the mechanism which rescue particular fractions of cells from drug-induced death requires analysis at the single cell level. We demonstrate the ability to assay the response of normal and CML stem cells to dasatinib, a small molecule drug approved for treatment of imatinib resistant CML, within a novel microfluidic platform. Dynamic, on-chip three-color viability assays reveal that difference in responses of normal and CML cell to dasatinib are observed even in the early phases of exposure, during which normal cells exhibit a significantly elevated cell death rate as compared to both controls and CML cells. However, even in the CML fraction, dasatinib markedly reduces cell migratory behaviors in all but five percent of cells. This resistant fraction may hold the key to understanding CML persistence following chemotherapeutic assault. This work demonstrates the potential utility of single cell microfluidic platform in stem cell biology

Nutritional intervention during gestation alters growth, body composition and gene expression patterns in skeletal muscle of pig offspring

peer-reviewedVariations in maternal nutrition during gestation can influence foetal growth, foetal development and permanently ‘programme’ offspring for postnatal life. The objective of this study was to analyse the effect of increased maternal nutrition during different gestation time windows on offspring growth, carcass quality, meat quality and gene expression in skeletal muscle. A total of 64 sows were assigned to the following feeding treatments: a standard control diet at a feed allocation of 2.3 kg/day throughout gestation, increased feed allowance of 4.6 kg/day from 25 to 50 days of gestation (dg), from 50 to 80 dg and from 25 to 80 dg. At weaning, Light, Medium and Heavy pigs of the same gender, within litter, were selected based on birth weight, individually penned and monitored until slaughter at 130 days post weaning. Carcass and meat quality traits of the semimembranosus (SM) muscle were recorded post mortem. A cross section of the semitendinosus (ST) muscle encompassing the deep and superficial regions were harvested from pigs (n518 per treatment) for RNA extraction and quantification of gene expression by real-time PCR. The results showed that doubling the feed intake from 25 to 50 dg reduced offspring growth, carcass weight, intramuscular fat content and increased drip loss of the SM muscle. Interestingly, protein phosphatase 3 catalytic subunit – a-isoform, which codes for the transcription factor calcineurin, was upregulated in the ST muscle of offspring whose mothers received increased feed allowance from 25 to 50 dg. This may provide an explanation for the previous observed increases in Type IIa muscle fibres of these offspring. Increasing the maternal feed intake from 50 to 80 dg negatively impacted pig growth and carcass weight, but produced leaner male pigs. Extending the increased maternal feed intake from 25 to 80 dg had no effect on offspring over the standard control gestation diet. Although intra-litter variation in pig weight is a problem for pig producers, increased maternal feeding offered no improvement throughout life to the lighter birth weight littermates in our study. Indeed, increased maternal nutrition at the three-gestation time windows selected provided no major benefits to the offspring.Teagasc, under the National Development Plan; Teagasc Walsh Fellowship; Short Term Scientific Mission (STSM) fund, COST925