High-throughput and sensitive quantitation of plasma catecholamines by ultraperformance liquid chromatography-tandem mass spectrometry using a solid phase microwell extraction plate.

Plasma catecholamines provide a reliable biomarker of sympathetic activity. The low circulating concentrations of catecholamines and analytical interferences require tedious sample preparation and long chromatographic runs to ensure their accurate quantification by HPLC with electrochemical detection. Published or commercially available methods relying on solid phase extraction technology lack sensitivity or require derivatization of catecholamine by hazardous reagents prior to tandem mass spectrometry (MS) analysis. Here, we manufactured a novel 96-well microplate device specifically designed to extract plasma catecholamines prior to their quantification by a new and highly sensitive ultraperformance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method. Processing time, which included sample purification on activated aluminum oxide and elution, is less than 1 h per 96-well microplate. The UPLC-MS/MS analysis run time is 2.0 min per sample. This UPLC-MS/MS method does not require a derivatization step, reduces the turnaround time by 10-fold compared to conventional methods used for routine application, and allows catecholamine quantification in reduced plasma sample volumes (50-250 μL, e.g., from children and mice).

Estudi de la viabilitat econòmica de la implantació d'una planta de piròlisi per a la producció de biochar en un context local i comarcal

En aquest document es pretén estudiar de la viabilitat econòmica de la implantació d’una planta de piròlisi per a la producció de biochar o char en un context local i comarcal. La biomassa és una font d’energia que genera uns rendiments energètics prou interesants i d’una manera respectuosa amb el medi ambient. Així doncs, la teòrica planta utilitzarà biomassa que, a traves del tractament termoquímic de la piròlisi, ens generarà uns productes que són d’utilitat per l’obtenció d’energia d’una manera respectuosa amb el medi ambient. Abans de fer l’anàlisi econòmic, hi ha una explicació extensa dels processos termoquímics, com són la piròlisi, la gasificació i la torrefacció. Posteriorment, es du a terme una revisió de l’estat actual de les tecnologies de conversió de biomassa a Catalunya i finalment es realitza un inventari i anàlisi dels usos de la biomassa a Catalunya, en el qual es parla principalment de l’estella el pèl·let, així com també de la seva producció, consum i exportació. El nostre anàlisi econòmic de la planta de processament de biomassa es durà a terme a nivell local, és a dir en un municipi, i també a nivell comarcal a Catalunya. A partir del processament de biomassa per mitja de la piròlisi, obtindrem uns productes, entre els quals el biochar, que serà un dels productes finals per poder vendre. L’altre producte serà el pèl·let de biochar, que l’obtindrem a través del procés de pel·letització. Aquest procés ens permetrà densificar el mateix biochar i obtenir-ne pèl·lets amb un poder calorífic superior, com també el seu possible preu de venda final. En aquest anàlisi econòmic plantejarem diferents escenaris tant a nivell local com comarcal, és a dir, farem variar diferents paràmetres de la planta, com poden ser els dies, les hores de treball, el sou dels treballadors, l’existència de procés de pel·letització...per veure la viabilitat del nostre projecte. Aquesta viabilitat la mesurarem amb diferents Índexs, entre els quals hi ha l’Índex de Rendibilitat, que ens determinarà si el projecte és possible si el seu valor és més gran a 1.

Biofuel Co-Product Uses for Pavement Geo-Materials Stabilization, TR-582, 2010

The production and use of biofuels has increased in the present context of sustainable development. Biofuel production from plant biomass produces not only biofuel or ethanol but also co-products containing lignin, modified lignin, and lignin derivatives. This research investigated the utilization of lignin-containing biofuel co-products (BCPs) in pavement soil stabilization as a new application area. Laboratory tests were conducted to evaluate the performance and the moisture susceptibility of two types of BCP-treated soil samples compared to the performance of untreated and traditional stabilizer-treated (fly ash) soil samples. The two types of BCPs investigated were (1) a liquid type with higher lignin content (co-product A) and (b) a powder type with lower lignin content (co-product B). Various additive combinations (co-product A and fly ash, co-products A and B, etc.) were also evaluated as alternatives to stand-alone co-products. Test results indicate that BCPs are effective in stabilizing the Iowa Class 10 soil classified as CL or A-6(8) and have excellent resistance to moisture degradation. Strengths and moisture resistance in comparison to traditional additives (fly ash) could be obtained through the use of combined additives (co-product A + fly ash; co-product A + co-product B). Utilizing BCPs as a soil stabilizer appears to be one of the many viable answers to the profitability of the bio-based products and the bioenergy business. Future research is needed to evaluate the freeze-thaw durability and for resilient modulus characterization of BCP-modified layers for a variety of pavement subgrade and base soil types. In addition, the long-term performance of these BCPs should be evaluated under actual field conditions and traffic loadings. Innovative uses of BCP in pavement-related applications could not only provide additional revenue streams to improve the economics of biorefineries, but could also serve to establish green road infrastructures.

Variation in biomass related variables of reed canary grass

Selostus: Ruokohelven biomassan tuotantoon vaikuttavien ominaisuuksien vaihtelu

Harvesting and transport of root biomass from fast-growing poplar plantations

Abstract

Alveolar epithelial CNGA1 channels mediate cGMP-stimulated, amiloride-insensitive, lung liquid absorption.

Impairment of lung liquid absorption can lead to severe respiratory symptoms, such as those observed in pulmonary oedema. In the adult lung, liquid absorption is driven by cation transport through two pathways: a well-established amiloride-sensitive Na(+) channel (ENaC) and, more controversially, an amiloride-insensitive channel that may belong to the cyclic nucleotide-gated (CNG) channel family. Here, we show robust CNGA1 (but not CNGA2 or CNGA3) channel expression principally in rat alveolar type I cells; CNGA3 was expressed in ciliated airway epithelial cells. Using a rat in situ lung liquid clearance assay, CNG channel activation with 1 mM 8Br-cGMP resulted in an approximate 1.8-fold stimulation of lung liquid absorption. There was no stimulation by 8Br-cGMP when applied in the presence of either 100 μM L: -cis-diltiazem or 100 nM pseudechetoxin (PsTx), a specific inhibitor of CNGA1 channels. Channel specificity of PsTx and amiloride was confirmed by patch clamp experiments showing that CNGA1 channels in HEK 293 cells were not inhibited by 100 μM amiloride and that recombinant αβγ-ENaC were not inhibited by 100 nM PsTx. Importantly, 8Br-cGMP stimulated lung liquid absorption in situ, even in the presence of 50 μM amiloride. Furthermore, neither L: -cis-diltiazem nor PsTx affected the β(2)-adrenoceptor agonist-stimulated lung liquid absorption, but, as expected, amiloride completely ablated it. Thus, transport through alveolar CNGA1 channels, located in type I cells, underlies the amiloride-insensitive component of lung liquid reabsorption. Furthermore, our in situ data highlight the potential of CNGA1 as a novel therapeutic target for the treatment of diseases characterised by lung liquid overload.

Short-term stability of testosterone and epitestosterone conjugates in urine samples: quantification by liquid chromatography-linear ion trap mass spectrometry.

A simple method using liquid chromatography-linear ion trap mass spectrometry for simultaneous determination of testosterone glucuronide (TG), testosterone sulfate (TS), epitestosterone glucuronide (EG) and epitestosterone sulfate (ES) in urine samples was developed. For validation purposes, a urine containing no detectable amount of TG, TS and EG was selected and fortified with steroid conjugate standards. Quantification was performed using deuterated testosterone conjugates to correct for ion suppression/enhancement during ESI. Assay validation was performed in terms of lower limit of detection (1-3ng/mL), recovery (89-101%), intraday precision (2.0-6.8%), interday precision (3.4-9.6%) and accuracy (101-103%). Application of the method to short-term stability testing of urine samples at temperature ranging from 4 to 37 degrees C during a time-storage of a week lead to the conclusion that addition of sodium azide (10mg/mL) is required for preservation of the analytes.

Fusion of the extended modified liquid drop model for nucleation and dynamical nucleation theory

We present a new phenomenological approach to nucleation, based on the combination of the extended modified liquid drop model and dynamical nucleation theory. The new model proposes a new cluster definition, which properly includes the effect of fluctuations, and it is consistent both thermodynamically and kinetically. The model is able to predict successfully the free energy of formation of the critical nucleus, using only macroscopic thermodynamic properties. It also accounts for the spinodal and provides excellent agreement with the result of recent simulations.

Elephant grass genotypes for bioenergy production by direct biomass combustion

The objective of this work was to evaluate elephant grass (Pennisetum purpureum Schum.) genotypes for bioenergy production by direct biomass combustion. Five elephant grass genotypes grown in two different soil types, both of low fertility, were evaluated. The experiment was carried out at Embrapa Agrobiologia field station in Seropédica, RJ, Brazil. The design was in randomized complete blocks, with split plots and four replicates. The genotypes studied were Cameroon, Bag 02, Gramafante, Roxo and CNPGL F06-3. Evaluations were made for biomass production, total biomass nitrogen, biomass nitrogen from biological fixation, carbon/nitrogen and stem/leaf ratios, and contents of fiber, lignin, cellulose and ash. The dry matter yields ranged from 45 to 67 Mg ha-1. Genotype Roxo had the lowest yield and genotypes Bag 02 and Cameroon had the highest ones. The biomass nitrogen accumulation varied from 240 to 343 kg ha-1. The plant nitrogen from biological fixation was 51% in average. The carbon/nitrogen and stem/leaf ratios and the contents of fiber, lignin, cellulose and ash did not vary among the genotypes. The five genotypes are suitable for energy production through combustion.

Variability of voriconazole plasma levels measured by new high-performance liquid chromatography and bioassay methods, [and] Voriconazole therapeutic drug monitoring in patients with invasive mycoses improves efficacy and safety outcomes

Rapport de synthèse1. Partie de laboratoireCette première étude décrit le développement et la validation, selon les standards internationaux, de deux techniques de mesure des concentrations sanguines de voriconazole, un nouvel agent antifongique à large spectre: 1) la chromatographic en phase liquide à haute pression et 2) le bio-essai utilisant une souche mutante de Candida hypersensible au voriconazole. Ce travail a aussi permis de mettre en évidence une importante et imprévisible variabilité inter- et intra-individuelle des concentrations sanguines de voriconazole malgré l'utilisation des doses recommandées par le fabriquant. Ce travail a été publié dans un journal avec "peer-review": "Variability of voriconazole plasma levels measured by new high- performance liquid chromatography and bioassay methods" by A. Pascual, V. Nieth, T. Calandra, J. Bille, S. Bolay, L.A. Decosterd, T. Buclin, P.A. Majcherczyk, D. Sanglard, 0. Marchetti. Antimicrobial Agents Chemotherapy, 2007; 51:137-432. Partie CliniqueCette deuxième étude a évalué de façon prospective l'impact clinique des concentrations sanguines de voriconazole sur l'efficacité et sécurité thérapeutique chez des patients atteints d'infections fongiques. Des concentrations sanguines élevées étaient significativement associés à la survenue d'une toxicité neurologique (encéphalopathie avec confusion, hallucinations et myoclonies) et des concentrations sanguines basses à une réponse insuffisante au traitement antifongique (persistance ou progression des signes cliniques et radiologiques de l'infection). Dans la majorité des cas, un ajustement de la dose de voriconazole, sur la base des concentrations mesurées, a abouti à une récupération neurologique complète ou à une résolution de l'infection, respectivement. Ce travail a été publié dans un journal avec "peer-review": " Voriconazole Therapeutic Drug Monitoring in Patients with Invasive Mycoses Improves Efficacy and Safety Outcomes" by A. Pascual, T. Calandra, S. Bolay, T. Buclin, J. Bille, and O. Marchetti. Clinical Infectious Diseases, 2008 January 15; 46(2): 201-11.Ces deux études, financées de façon conjointe par un "grant" international de la Société suisse d'infectiologie et la Société internationale de maladies infectieuses et par la Fondation pour le progrès en microbiologie médicale et maladies infectieuses (FAMMID, Lausanne), ont été réalisées au sein du Service des Maladies Infectieuses, Département de Médecine, au CHUV, en étroite collaboration avec la Division de Pharmacologie Clinique, Département de Médecine, au CHUV et l'Institut de Microbiologie du CHUV et de l'Université de Lausanne.

Tropical terrestrial model ecosystems for evaluation of soil fauna and leaf litter quality effects on litter consumption, soil microbial biomass and plant growth

The aim of this work was to evaluate whether terrestrial model ecosystems (TMEs) are a useful tool for the study of the effects of litter quality, soil invertebrates and mineral fertilizer on litter decomposition and plant growth under controlled conditions in the tropics. Forty-eight intact soil cores (17.5-cm diameter, 30-cm length) were taken out from an abandoned rubber plantation on Ferralsol soil (Latossolo Amarelo) in Central Amazonia, Brazil, and kept at 28ºC in the laboratory during four months. Leaf litter of either Hevea pauciflora (rubber tree), Flemingia macrophylla (a shrubby legume) or Brachiaria decumbens (a pasture grass) was put on top of each TME. Five specimens of either Pontoscolex corethrurus or Eisenia fetida (earthworms), Porcellionides pruinosus or Circoniscus ornatus (woodlice), and Trigoniulus corallinus (millipedes) were then added to the TMEs. Leaf litter type significantly affected litter consumption, soil microbial biomass and nitrate concentration in the leachate of all TMEs, but had no measurable effect on the shoot biomass of rice seedlings planted in top soil taken from the TMEs. Feeding rates measured with bait lamina were significantly higher in TMEs with the earthworm P. corethrurus and the woodlouse C. ornatus. TMEs are an appropriate tool to assess trophic interactions in tropical soil ecossistems under controlled laboratory conditions.

Detection of metabolite induction in fungal co-cultures on solid media by high-throughput differential ultra-high pressure liquid chromatography-time-of-flight mass spectrometry fingerprinting.

Access to new biological sources is a key element of natural product research. A particularly large number of biologically active molecules have been found to originate from microorganisms. Very recently, the use of fungal co-culture to activate the silent genes involved in metabolite biosynthesis was found to be a successful method for the induction of new compounds. However, the detection and identification of the induced metabolites in the confrontation zone where fungi interact remain very challenging. To tackle this issue, a high-throughput UHPLC-TOF-MS-based metabolomic approach has been developed for the screening of fungal co-cultures in solid media at the petri dish level. The metabolites that were overexpressed because of fungal interactions were highlighted by comparing the LC-MS data obtained from the co-cultures and their corresponding mono-cultures. This comparison was achieved by subjecting automatically generated peak lists to statistical treatments. This strategy has been applied to more than 600 co-culture experiments that mainly involved fungal strains from the Fusarium genera, although experiments were also completed with a selection of several other filamentous fungi. This strategy was found to provide satisfactory repeatability and was used to detect the biomarkers of fungal induction in a large panel of filamentous fungi. This study demonstrates that co-culture results in consistent induction of potentially new metabolites.

Analysis of cannabinoids in oral fluid by liquid chromatography-tandem mass spectrometry

A sensitive method was developed for quantifying a wide range of cannabinoids in oral fluid (OF) by liquid chromatography-tandem mass spectrometry (LC-MS/MS). These cannabinoids include a dagger(9)-tetrahydrocannabinol (THC), 11-hydroxy-a dagger(9)-tetrahydrocannabinol (11-OH-THC), 11-nor-9-carboxy-a dagger(9)-tetrahydrocannabinol (THCCOOH), cannabinol (CBN), cannabidiol (CBD), a dagger(9)-tetrahydrocannabinolic acid A (THC-A), 11-nor-9-carboxy-a dagger(9)-tetrahydrocannabinol glucuronide (THCCOOH-gluc), and a dagger(9)-tetrahydrocannabinol glucuronide (THC-gluc). Samples were collected using a Quantisal (TM) device. The advantages of performing a liquid-liquid extraction (LLE) of KCl-saturated OF using heptane/ethyl acetate versus a solid-phase extraction (SPE) using HLB copolymer columns were determined. Chromatographic separation was achieved in 11.5 min on a Kinetex (TM) column packed with 2.6-mu m core-shell particles. Both positive (THC, 11-OH-THC, CBN, and CBD) and negative (THCCOOH, THC-gluc, THCCOOH-gluc, and THC-A) electrospray ionization modes were employed with multiple reaction monitoring using a high-end AB Sciex API 5000 (TM) triple quadrupole LC-MS/MS system. Unlike SPE, LLE failed to extract THC-gluc and THCCOOH-gluc. However, the LLE method was more sensitive for the detection of THCCOOH than the SPE method, wherein the limit of detection (LOD) and limit of quantification (LOQ) decreased from 100 to 50 pg/ml and from 500 to 80 pg/ml, respectively. The two extraction methods were successfully applied to OF samples collected from volunteers before and after they smoked a homemade cannabis joint. High levels of THC were measured soon after smoking, in addition to significant amounts of THC-A. Other cannabinoids were found in low concentrations. Glucuronide conjugate levels were lower than the method's LOD for most samples. Incubation studies suggest that glucuronides could be enzymatically degraded by glucuronidase prior to OF collection

Letter health consultation : Exposure to benzene from Equistar pyrolysis gasoline tank, Clinton, Iowa (2008)

The Iowa Department of Public Health (IDPH) has been asked by several citizens of Clinton, Iowa to evaluate potential health effects from exposure to benzene from an above ground tank located near a residential area. The above ground tank is managed by Equistar Chemicals, LP (Equistar) and is use to store pyrolysis gasoline prior to off-loading onto barges. The IDPH in cooperation with the Agency for Toxic Substances and Disease Registry (ATSDR) prepared this health consultation to review the air quality monitoring provided by Equistar and provide an evaluation of the health effects from exposure to benzene at the levels detected in ambient air near the tank. The information in this health consultation was current at the time of writing. Data that emerges later could alter this document’s conclusions and recommendations.

Combining phospholipases and a liquid lipase for one-step biodiesel production using crude oils

Background Enzymatic biodiesel is becoming an increasingly popular topic in bioenergy literature because of its potential to overcome the problems posed by chemical processes. However, the high cost of the enzymatic process still remains the main drawback for its industrial application, mostly because of the high price of refined oils. Unfortunately, low cost substrates, such as crude soybean oil, often release a product that hardly accomplishes the final required biodiesel specifications and need an additional pretreatment for gums removal. In order to reduce costs and to make the enzymatic process more efficient, we developed an innovative system for enzymatic biodiesel production involving a combination of a lipase and two phospholipases. This allows performing the enzymatic degumming and transesterification in a single step, using crude soybean oil as feedstock, and converting part of the phospholipids into biodiesel. Since the two processes have never been studied together, an accurate analysis of the different reaction components and conditions was carried out. Results Crude soybean oil, used as low cost feedstock, is characterized by a high content of phospholipids (900 ppm of phosphorus). However, after the combined activity of different phospholipases and liquid lipase Callera Trans L, a complete transformation into fatty acid methyl esters (FAMEs >95%) and a good reduction of phosphorus (P <5 ppm) was achieved. The combination of enzymes allowed avoidance of the acid treatment required for gums removal, the consequent caustic neutralization, and the high temperature commonly used in degumming systems, making the overall process more eco-friendly and with higher yield. Once the conditions were established, the process was also tested with different vegetable oils with variable phosphorus contents. Conclusions Use of liquid lipase Callera Trans L in biodiesel production can provide numerous and sustainable benefits. Besides reducing the costs derived from enzyme immobilization, the lipase can be used in combination with other enzymes such as phospholipases for gums removal, thus allowing the use of much cheaper, non-refined oils. The possibility to perform degumming and transesterification in a single tank involves a great efficiency increase in the new era of enzymatic biodiesel production at industrial scale.