Investigation of probe substrates to assess hepatic transport function

The objective of this research project was to identify and characterize probe substrates for specific hepatic transport proteins. Identification of specific probe substrates for basolateral and canalicular transport proteins is necessary to elucidate mechanisms of hepatobiliary drug transport, phenotype for interindividual differences in transport protein function, and identify potential drug-drug interactions at the hepatic transport level. Three probe substrates were selected for investigation: fexofenadine, 99mTechnetium mebrofenin (99mTc-MEB), and 99mTechnetium sestamibi (99mTc-MIBI). Although fexofenadine has been touted as a P-gp specific probe substrate, pharmacokinetic modeling/siumulation of clinical data, sandwich-cultured human hepatocyte experiments and perfused liver studies in rats and mice demonstrated that fexofenadine is not a suitable probe for a specific transport protein due to compensatory efflux pathways. The hepatic uptake and excretion of 99mTc-MEB and 99mTc-MIBI have not been investigated fully despite their use as non-invasive probes to assess transport function. Therefore, suspended and sandwich-cultured rat and human hepatocytes were used to fully characterize the mechanisms of hepatic transport of 99mTc-MEB and 99mTc-MIBI. Then to validate the use of 99mTc-MEB and/or 99mTc-MIBI as probe substrates to predict the hepatic clearance of the anticancer agent, sorafenib studies were conducted to confirm similar mechanisms of hepatic uptake. Lastly, the pharmacokinetics and hepatic exposure of 99mTc-MIBI and 99mTc-MEB were compared in a patient with hepatocellular carcinoma and Child's Pugh B cirrhosis vs. healthy human volunteers. Pharmacokinetic models were constructed to describe the distribution and elimination of 99mTc-MEB and 99mTc-MIBI, to compare alterations in key rate constants representing hepatic uptake and efflux mechanisms secondary to disease. In conclusion, 99mTc-MEB and 99mTc-MIBI may be very useful phenotypic probes that are sensitive to changes in hepatic function associated with liver disease. The hepatic exposure to 99mTc-MIBI was decreased in the patient with hepatocellular carcinoma and cirrhosis compared to healthy volunteers, whereas the hepatic exposure of 99mTc-MEB was similar but the hepatic exposure profile was notably different with a lower maximum but more prolonged exposure. Collectively, the results of this research negate the use of fexofenadine as a probe for hepatic transport, but support the use of 99mTc-MEB and 99mTc-MIBI as probe substrates, which add the unique ability to quantify liver concentrations

Turbulent Driving Scales in Molecular Clouds

Supersonic turbulence in molecular clouds is a dominant agent that strongly affects the clouds' evolution and star formation activity. Turbulence may be initiated and maintained by a number of processes, acting at a wide range of physical scales. By examining the dynamical state of molecular clouds, it is possible to assess the primary candidates for how the turbulent energy is injected. The aim of this paper is to constrain the scales at which turbulence is driven in the molecular interstellar medium, by comparing simulated molecular spectral line observations of numerical magnetohydrodynamic (MHD) models and molecular spectral line observations of real molecular clouds. We use principal component analysis, applied to both models and observational data, to extract a quantitative measure of the driving scale of turbulence. We find that only models driven at large scales (comparable to, or exceeding, the size of the cloud) are consistent with observations. This result applies also to clouds with little or no internal star formation activity. Astrophysical processes acting on large scales, including supernova-driven turbulence, magnetorotational instability, or spiral shock forcing, are viable candidates for the generation and maintenance of molecular cloud turbulence. Small scale driving by sources internal to molecular clouds, such as outflows, can be important on small scales, but cannot replicate the observed large-scale velocity fluctuations in the molecular interstellar medium.Comment: 8 pages, 7 figures, accepted for publication in A&

Managing soil fertility in organic farming systems

Complex relationships exist between different components of the organic farm and the quantity and quality of the end products depend on the functioning of the whole system. As such, it is very difficult to isolate soil fertility from production and environmental aspects of the system. Crop rotation is the central tool that integrates the maintenance and development of soil fertility with different aspects of crop and livestock production in organic systems. Nutrient supply to crops depends on the use of legumes to add nitrogen to the system and limited inputs of supplementary nutrients, added in acceptable forms. Manures and crop residues are carefully managed to recycle nutrients around the farm. Management of soil organic matter, primarily through the use of short-term leys, helps ensure good soil structure and biological activity, important for nutrient supply, health and productivity of both crops and livestock. Carefully planned diverse rotations help reduce the incidence of pests and diseases and allow for cultural methods of weed control. As a result of the complex interactions between different system components, fertility management in organic farming relies on a long-term integrated approach rather than the more short-term very targeted solutions common in conventional agriculture

Next-generation conservation genetics and biodiversity monitoring

This special issue of Evolutionary Applications consists of 10 publications investigating the use of next-generation tools and techniques in population genetic analyses and biodiversity assessment. The special issue stems from a 2016 Next Generation Genetic Monitoring Workshop, hosted by the National Institute for Mathematical and Biological Synthesis (NIMBioS) in Tennessee, USA. The improved accessibility of next-generation sequencing platforms has allowed molecular ecologists to rapidly produce large amounts of data. However, with the increased availability of new genomic markers and mathematical techniques, care is needed in selecting appropriate study designs, interpreting results in light of conservation concerns, and determining appropriate management actions. This special issue identifies key attributes of successful genetic data analyses in biodiversity evaluation and suggests ways to improve analyses and their application in current population and conservation genetics research

ATM variants 7271T>G and IVS10-6T>G among women with unilateral and bilateral breast cancer

Recent reports suggest that two ATM gene mutations, 7271T>G and IVS10-6T>G, are associated with a high risk of breast cancer among multiple-case families. To assess the importance of these two mutations in another 'high-risk' group, young women (under age 51) with multiple primaries, we screened a large population-based series of young women with bilateral breast cancer and compared the frequency of these mutations among similar women diagnosed with unilateral breast cancer. The 1149 women included were enrolled in an ongoing population-based case-control study of the genetic factors that contribute to bilateral breast cancer; they were not selected on the basis of family history of cancer. Screening for 7271T>G and IVS10-6T>G ATM gene mutations was conducted using DHPLC followed by direct sequencing. The 7271T>G mutation was detected in one out of 638 (0.2%) women with unilateral breast cancer and in none of the bilateral cases, and the IVS10-6T>G mutation in one out of 511 (0.2%) bilateral and in eight out of 638 (1.3%) unilateral breast cancer cases. Carriers of either mutation were not limited to women with a family history. Given the likelihood that young women with bilateral breast cancer have a genetic predisposition, the observed mutation distribution is contrary to that expected if these two mutations were to play an important role in breast carcinogenesis among individuals at high risk

ATM variants and cancer risk in breast cancer patients from Southern Finland

BACKGROUND: Individuals heterozygous for germline ATM mutations have been reported to have an increased risk for breast cancer but the role for ATM genetic variants for breast cancer risk has remained unclear. Recently, a common ATM variant, ATMivs38 -8T>C in cis with the ATMex39 5557G>A (D1853N) variant, was suggested to associate with bilateral breast cancer among familial breast cancer patients from Northern Finland. We have here evaluated the 5557G>A and ivs38-8T>C variants in an extensive case-control association analysis. We also aimed to investigate whether there are other ATM mutations or variants contributing to breast cancer risk in our population. METHODS: Two common ATM variants, 5557G>A and ivs38-8T>C, previously suggested to associate with bilateral breast cancer, were genotyped in an extensive set of 786 familial and 884 unselected breast cancer cases as well as 708 healthy controls. We also screened the entire coding region and exon-intron boundaries of the ATM gene in 47 familial breast cancer patients and constructed haplotypes of the patients. The identified variants were also evaluated for increased breast cancer risk among additional breast cancer cases and controls. RESULTS: Neither of the two common variants, 5557G>A and ivs38-8T>C, nor any haplotype containing them, was significantly associated with breast cancer risk, bilateral breast cancer or multiple primary cancers in any of the patient groups or subgoups. Three rare missense alterations and one intronic change were each found in only one patient of over 250 familial patients studied and not among controls. The fourth missense alteration studied further was found with closely similar frequencies in over 600 familial cases and controls. CONCLUSION: Altogether, our results suggest very minor effect, if any, of ATM genetic variants on familial breast cancer in Southern Finland. Our results do not support association of the 5557G>A or ivs38-8T>C variant with increased breast cancer risk or with bilateral breast cancer

Breast and other cancers in 1445 blood relatives of 75 Nordic patients with ataxia telangiectasia

Epidemiological studies have consistently shown elevated rates of breast cancer among female blood relatives of patients with ataxia telangiectasia (AT), a rare autosomal recessive disease. A large proportion of the members of AT families are carriers of AT-causing gene mutations in ATM (Ataxia Telangiectasia Mutated), and it has been hypothesised that these otherwise healthy carriers are predisposed to breast cancer. This is an extended and enlarged follow-up study of cancer incidence in blood relatives of 75 patients with verified AT in 66 Nordic families. Blood relatives were identified through population registry linkages, and the occurrence of cancer was determined from cancer registry files in each country and compared with national incidence rates. The ATM mutation carrier probabilities of relatives were assigned from the combined information on location in family, consanguinity, if any, and supplementary carrier screening in some families. Among the 1445 blood relatives of AT patients, 225 cancers were observed, with 170.4 expected, yielding a standardised incidence ratio (SIR) of 1.3 (95% confidence interval (CI), 1.1–1.4). Invasive breast cancer occurred in 34 female relatives (SIR, 1.7; 95% CI, 1.2–2.4) and was diagnosed in 21 women before the age of 55 years (SIR, 2.9; 95% CI, 1.8–4.5), including seven mothers of probands (SIR, 8.1; 95% CI, 3.3–17). When the group of mothers was excluded, no clear relationship was observed between the allocated mutation carrier probability of each family member and the extent of breast cancer risk. We concluded that the increased risk for female breast cancer seen in 66 Nordic AT families appeared to be restricted to women under the age of 55 years and was due mainly to a very high risk in the group of mothers. The findings of breast cancer risk in mothers, but not other likely mutation carriers, in this and other studies raises questions about the hypothesis of a simple causal relationship with ATM heterozygosity

Linkage disequilibrium pattern of the ATM gene in breast cancer patients and controls; association of SNPs and haplotypes to radio-sensitivity and post-lumpectomy local recurrence

<p>Abstract</p> <p>Background</p> <p>The ATM protein is activated as a result of ionizing radiation, and genetic variants of the <it>ATM </it>gene may therefore affect the level of radiation-induced damage. Individuals heterozygous for <it>ATM </it>mutations have been reported to have an increased risk of malignancy, especially breast cancer.</p> <p>Materials and methods</p> <p>Norwegian breast cancer patients (272) treated with radiation (252 of which were evaluated for radiation-induced adverse side effects), 95 Norwegian women with no known history of cancer and 95 American breast cancer patients treated with radiation (44 of which developed ipsilateral breast tumour recurrence, IBTR) were screened for sequence variations in all exons of the <it>ATM </it>gene as well as known intronic variants by denaturating high performance liquid chromatography (dHPLC) followed by sequencing to determine the nature of the variant.</p> <p>Results and Conclusion</p> <p>A total of 56 variants were identified in the three materials combined. A borderline significant association with breast cancer risk was found for the 1229 T>C (Val>Ala) substitution in exon 11 (P-value 0.055) between the Norwegian controls and breast cancer patients as well as a borderline significant difference in haplotype distribution (P-value 0.06). Adverse side effects, such as: development of costal fractures and telangiectasias, subcutaneous and lung fibrosis, pleural thickening and atrophy were evaluated in the Norwegian patients. Significant associations were found for several of the identified variants such as rs1800058 (Leu > Phe) where a decrease in minor allele frequency was found with increasing level of adverse side effects for the clinical end-points pleural thickening and lung fibrosis, thus giving a protective effect. Overall our results indicate a role for variation in the <it>ATM </it>gene both for risk of developing breast cancer, and in radiation induced adverse side effects. No association could be found between risk of developing ipsilateral breast tumour recurrence and any of the sequence variants found in the American patient material.</p

Continuum-based models and concepts for the transport of nanoparticles in saturated porous media: A state-of-the-science review

Environmental applications of nanoparticles (NP) increasingly result in widespread NP distribution within porous media where they are subject to various concurrent transport mechanisms including irreversible deposition, attachment/detachment (equilibrium or kinetic), agglomeration, physical straining, site-blocking, ripening, and size exclusion. Fundamental research in NP transport is typically conducted at small scale, and theoretical mechanistic modeling of particle transport in porous media faces challenges when considering the simultaneous effects of transport mechanisms. Continuum modeling approaches, in contrast, are scalable across various scales ranging from column experiments to aquifer. They have also been able to successfully describe the simultaneous occurrence of various transport mechanisms of NP in porous media such as blocking/straining or agglomeration/deposition/detachment. However, the diversity of model equations developed by different authors and the lack of effective approaches for their validation present obstacles to the successful robust application of these models for describing or predicting NP transport phenomena. This review aims to describe consistently all the important NP transport mechanisms along with their representative mathematical continuum models as found in the current scientific literature. Detailed characterizations of each transport phenomenon in regards to their manifestation in the column experiment outcomes, i.e., breakthrough curve (BTC) and residual concentration profile (RCP), are presented to facilitate future interpretations of BTCs and RCPs. The review highlights two NP transport mechanisms, agglomeration and size exclusion, which are potentially of great importance in controlling the fate and transport of NP in the subsurface media yet have been widely neglected in many existing modeling studies. A critical limitation of the continuum modeling approach is the number of parameters used upon application to larger scales and when a series of transport mechanisms are involved. We investigate the use of simplifying assumptions, such as the equilibrium assumption, in modeling the attachment/detachment mechanisms within a continuum modelling framework. While acknowledging criticisms about the use of this assumption for NP deposition on a mechanistic (process) basis, we found that its use as a description of dynamic deposition behavior in a continuum model yields broadly similar results to those arising from a kinetic model. Furthermore, we show that in two dimensional (2-D) continuum models the modeling efficiency based on the Akaike information criterion (AIC) is enhanced for equilibrium vs kinetic with no significant reduction in model performance. This is because fewer parameters are needed for the equilibrium model compared to the kinetic model. Two major transport regimes are identified in the transport of NP within porous media. The first regime is characterized by higher particle-surface attachment affinity than particle-particle attachment affinity, and operative transport mechanisms of physicochemical filtration, blocking, and physical retention. The second regime is characterized by the domination of particle-particle attachment tendency over particle-surface affinity. In this regime although physicochemical filtration as well as straining may still be operative, ripening is predominant together with agglomeration and further subsequent retention. In both regimes careful assessment of NP fate and transport is necessary since certain combinations of concurrent transport phenomena leading to large migration distances are possible in either case