LC-MS/MS method for simultaneous analysis of uracil, 5,6-dihydrouracil, 5-fluorouracil and 5-fluoro-5,6-dihydrouracil in human plasma for therapeutic drug monitoring and toxicity prediction in cancer patients

The chemotherapeutic drug 5-fluorouracil (5-FU) is widely used for treating solid tumors. Response to 5-FU treatment is variable with 10-30% of patients experiencing serious toxicity partly explained by reduced activity of dihydropyrimidine dehydrogenase (DPD). DPD converts endogenous uracil (U) into 5,6-dihydrouracil (UH(2) ), and analogously, 5-FU into 5-fluoro-5,6-dihydrouracil (5-FUH(2) ). Combined quantification of U and UH(2) with 5-FU and 5-FUH(2) may provide a pre-therapeutic assessment of DPD activity and further guide drug dosing during therapy. Here, we report the development of a liquid chromatography-tandem mass spectrometry assay for simultaneous quantification of U, UH(2) , 5-FU and 5-FUH(2) in human plasma. Samples were prepared by liquid-liquid extraction with 10:1 ethyl acetate-2-propanol (v/v). The evaporated samples were reconstituted in 0.1% formic acid and 10 μL aliquots were injected into the HPLC system. Analyte separation was achieved on an Atlantis dC(18) column with a mobile phase consisting of 1.0 mm ammonium acetate, 0.5 mm formic acid and 3.3% methanol. Positively ionized analytes were detected by multiple reaction monitoring. The analytical response was linear in the range 0.01-10 μm for U, 0.1-10 μm for UH(2) , 0.1-75 μm for 5-FU and 0.75-75 μm for 5-FUH(2) , covering the expected concentration ranges in plasma. The method was validated following the FDA guidelines and applied to clinical samples obtained from ten 5-FU-treated colorectal cancer patients. The present method merges the analysis of 5-FU pharmacokinetics and DPD activity into a single assay representing a valuable tool to improve the efficacy and safety of 5-FU-based chemotherapy.

The GABRIEL Advanced Surveys: study design, participation and evaluation of bias

Exposure to farming environments has been shown to protect substantially against asthma and atopic disease across Europe and in other parts of the world. The GABRIEL Advanced Surveys (GABRIELA) were conducted to determine factors in farming environments which are fundamental to protecting against asthma and atopic disease. The GABRIEL Advanced Surveys have a multi-phase stratified design. In a first-screening phase, a comprehensive population-based survey was conducted to assess the prevalence of exposure to farming environments and of asthma and atopic diseases (n = 103,219). The second phase was designed to ascertain detailed exposure to farming environments and to collect biomaterial and environmental samples in a stratified random sample of phase 1 participants (n = 15,255). A third phase was carried out in a further stratified sample only in Bavaria, southern Germany, aiming at in-depth respiratory disease and exposure assessment including extensive environmental sampling (n = 895). Participation rates in phase 1 were around 60% but only about half of the participating study population consented to further study modules in phase 2. We found that consenting behaviour was related to familial allergies, high parental education, wheeze, doctor diagnosed asthma and rhinoconjunctivitis, and to a lesser extent to exposure to farming environments. The association of exposure to farm environments with asthma or rhinoconjunctivitis was not biased by participation or consenting behaviour. The GABRIEL Advanced Surveys are one of the largest studies to shed light on the protective 'farm effect' on asthma and atopic disease. Bias with regard to the main study question was able to be ruled out by representativeness and high participation rates in phases 2 and 3. The GABRIEL Advanced Surveys have created extensive collections of questionnaire data, biomaterial and environmental samples promising new insights into this area of research.

Deforestation and forest degradation monitoring and assessment of biomass and carbon stock of lowland rainforest in the Analanjirofo region, Madagascar

Madagascar is currently developing a policy and strategies to enhance the sustainable management of its natural resources, encouraged by United Nations Framework Convention on Climate Change (UNFCCC) and REDD. To set up a sustainable financing scheme methodologies have to be provided that estimate, prevent and mitigate leakage, develop national and regional baselines, and estimate carbon benefits. With this research study this challenge was tried to be addressed by analysing a lowland rainforest in the Analanjirofo region in the district of Soanierana Ivongo, North East of Madagascar. For two distinguished forest degradation stages: “low degraded forest” and “degraded forest” aboveground biomass and carbon stock was assessed. The corresponding rates of carbon within those two classes were calculated and linked to a multi-temporal set of SPOT satellite data acquired in 1991, 2004 and 2009. Deforestation and particularly degradation and the related carbon stock developments were analysed. With the assessed data for the 3 years 1991, 2004 and 2009 it was possible to model a baseline and to develop a forest prediction for 2020 for Analanjirofo region in the district of Soanierana Ivongo. These results, developed applying robust methods, may provide important spatial information regarding the priorities in planning and implementation of future REDD+ activities in the area.

[Laboratory diagnostic with regard to new anticoagulants - monitoring and influence on coagulation tests]

New oral anticoagulants promise to overcome essential drawbacks of traditional substances. They have a predictable therapeutic effect, a wide therapeutic window, only limited interaction with food and drugs and can be administered p.o. with a fixed dose. On the other hand, knowledge on the laboratory management of new anticoagulants is limited. In the present article we discuss possible indications and available assays for monitoring of Rivaroxaban, Apixaban and Dabigatran. Furthermore, we discuss interpretation of routine coagulation tests during therapy with these new drugs.

Antimicrobial use in Swiss dairy farms: quantification and evaluation of data quality

Data on antimicrobial use play a key role in the development of policies for the containment of antimicrobial resistance. On-farm data could provide a detailed overview of the antimicrobial use, but technical and methodological aspects of data collection and interpretation, as well as data quality need to be further assessed. The aims of this study were (1) to quantify antimicrobial use in the study population using different units of measurement and contrast the results obtained, (2) to evaluate data quality of farm records on antimicrobial use, and (3) to compare data quality of different recording systems. During 1 year, data on antimicrobial use were collected from 97 dairy farms. Antimicrobial consumption was quantified using: (1) the incidence density of antimicrobial treatments; (2) the weight of active substance; (3) the used daily dose and (4) the used course dose for antimicrobials for intestinal, intrauterine and systemic use; and (5) the used unit dose, for antimicrobials for intramammary use. Data quality was evaluated by describing completeness and accuracy of the recorded information, and by comparing farmers' and veterinarians' records. Relative consumption of antimicrobials depended on the unit of measurement: used doses reflected the treatment intensity better than weight of active substance. The use of antimicrobials classified as high priority was low, although under- and overdosing were frequently observed. Electronic recording systems allowed better traceability of the animals treated. Recording drug name or dosage often resulted in incomplete or inaccurate information. Veterinarians tended to record more drugs than farmers. The integration of veterinarian and farm data would improve data quality.

[First detection of "Candidatus Mycoplasma haemolamae" in South American Camelids of Switzerland and evaluation of prevalence]

Haemotrophic mycoplasmas (also known as haemoplasmas), small bacterias which parasite the surface of erythrocytes, have been described in several species. Recently, molecular methods were developed for the diagnosis of haemoplasma infection. The presented study describes the first detection and the investigation of prevalence of "Candidatus Mycoplasma haemolamae" in South American Camelids in Switzerland. A random sample of the latter population was tested for haemoplasma infections using real-time PCR. The infection was detected in 18.6% of the animals and was found both in indigenous and in imported camelids. Of the tested herds 39,1% harboured at least one animal positive for haemoplasmas in PCR. There was no difference in prevalence between male and female animals and llamas and alpacas, respectively. Furthermore, the prevalence of infection was not significantly different in diseased animals compared to healthy camelids. From the latter observation and the fact that the high prevalence was accompanied by an undetectable incidence, we concluded that the pathogenicity of "Candidatus Mycoplasma haemolamae" may be low.

Incidence of descending aortic pathology and evaluation of the impact of thoracic endovascular aortic repair: a population-based study in England and Wales from 1999 to 2010

To investigate population trends in thoracic aortic disease (dissections and aneurysms) in England and Wales, with focus on the impact of thoracic endovascular aortic repair on procedure numbers and age at repair.

Monitoring and Modeling of the Hydraulic and Sediment Processes at In-Stream Restoration Structures in the Vicinity of a Bridge Crossing

The long-term performance of infrastructure depends on reliable and sustainable designs. Many of Pennsylvania’s streams experience sediment transport problems that increase maintenance costs and lower structural integrity of bridge crossings. A stream restoration project is one common mitigation measure used to correct such problems at bridge crossings. Specifically, in an attempt to alleviate aggradation problems with the Old Route 15 Bridge crossing on White Deer Creek, in White Deer, PA, two in-stream structures (rock cross vanes) and several bank stabilization features were installed along with a complete channel redevelopment. The objectives of this research were to characterize the hydraulic and sediment transport processes occurring at the White Deer Creek site, and to investigate, through physical and mathematical modeling, the use of instream restoration structures. The goal is to be able to use the results of this study to prevent aggradation or other sediment related problems in the vicinity of bridges through improved design considerations. Monitoring and modeling indicate that the study site on White Deer Creek is currently unstable, experiencing general channel down-cutting, bank erosion, and several local areas of increased aggradation and degradation of the channel bed. An in-stream structure installed upstream of the Old Route 15 Bridge failed by sediment burial caused by the high sediment load that White Deer Creek is transporting as well as the backwater effects caused by the bridge crossing. The in-stream structure installed downstream of the Old Route 15 Bridge is beginning to fail because of the alignment of the structure with the approach direction of flow from upstream of the restoration structure.

Characterization of the canine CLCN3 gene and evaluation as candidate for late-onset NCL

BACKGROUND: The neuronal ceroid lipofuscinoses (NCL) are a heterogenous group of inherited progressive neurodegenerative diseases in different mammalian species. Tibetan Terrier and Polish Owczarek Nizinny (PON) dogs show rare late-onset NCL variants with autosomal recessive inheritance, which can not be explained by mutations of known human NCL genes. These dog breeds represent animal models for human late-onset NCL. In mice the chloride channel 3 gene (Clcn3) encoding an intracellular chloride channel was described to cause a phenotype similar to NCL. RESULTS: Two full-length cDNA splice variants of the canine CLCN3 gene are reported. The current canine whole genome sequence assembly was used for gene structure analyses and revealed 13 coding CLCN3 exons in 52 kb of genomic sequence. Sequence analysis of the coding exons and flanking intron regions of CLCN3 using six NCL-affected Tibetan terrier dogs and an NCL-affected Polish Owczarek Nizinny (PON) dog, as well as eight healthy Tibetan terrier dogs revealed 13 SNPs. No consistent CLCN3 haplotype was associated with NCL. CONCLUSION: For the examined animals we excluded the complete coding region and adjacent intronic regions of canine CLCN3 to harbor disease-causing mutations. Therefore it seems to be unlikely that a mutation in this gene is responsible for the late-onset NCL phenotype in these two dog breeds.