Detection in urine of 4-methyl-2-hexaneamine, a doping agent.

Stimulants are banned in-competition for all categories of sports by the World Anti-Doping Agency. A simple liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay employing electrospray ionisation in positive mode was developed in that work for the quantification in urine specimens of 4-methyl-2-hexaneamine, a primary amine exhibiting sympathomimetic properties. Following a simple pretreatment procedure, the analyte was separated using a gradient mobile phase on reverse phase C8 column. Selected reaction monitoring m/z 116.2-->57.3 was specific for detection of 4-methyl-2-hexaneamine and the assay exhibited a linear dynamic range of 50-700 ng/mL. The validated method has been successfully applied to analyze the target compound in food supplements as well as in urine specimens. The administered drug (40 mg) was detected at the level of 350 ng/mL in the urine up to 4 days.

Comparison of cannabinoid concentrations in oral fluid and whole blood between occasional and regular cannabis smokers prior to and after smoking a cannabis joint.

A cross-over controlled administration study of smoked cannabis was carried out on occasional and heavy smokers. The participants smoked a joint (11 % Δ9-tetrahydrocannabinol (THC)) or a matching placebo on two different occasions. Whole blood (WB) and oral fluid (OF) samples were collected before and up to 3.5 h after smoking the joints. Pharmacokinetic analyses were obtained from these data. Questionnaires assessing the subjective effects were administered to the subjects during each session before and after the smoking time period. THC, 11-hydroxy-THC (11-OH-THC) and 11-nor-9-carboxy-THC (THCCOOH) were analyzed in the blood by gas chromatography or liquid chromatography (LC)-tandem mass spectrometry (MS/MS). The determination of THC, THCCOOH, cannabinol (CBN), and Δ9-tetrahydrocannabinolic acid A (THC-A) was carried out on OF only using LC-MS/MS. In line with the widely accepted assumption that cannabis smoking results in a strong contamination of the oral cavity, we found that THC, and also THC-A, shows a sharp, high concentration peak just after smoking, with a rapid decrease in these levels within 3 h. No obvious differences were found between both groups concerning THC median maximum concentrations measured either in blood or in OF; these levels were equal to 1,338 and 1,041 μg/L in OF and to 82 and 94 μg/L in WB for occasional and heavy smokers, respectively. The initial WB THCCOOH concentration was much higher in regular smokers than in occasional users. Compared with the occasional smokers, the sensation of confusion felt by the regular smokers was much less while the feeling of intoxication remained almost unchanged.

Mephedrone pharmacokinetics after intravenous and oral administration in rats: relation to pharmacodynamics

Rationale Mephedrone (4-methylmethcathinone) is a still poorly known drug of abuse, alternative to ecstasy or cocaine. Objective The major aims were to investigate the pharmacokineticsa and locomotor activity of mephedrone in rats and provide a pharmacokinetic/pharmacodynamic model. Methods Mephedrone was administered to male SpragueDawley rats intravenously (10 mg/kg) and orally (30 and 60 mg/kg). Plasma concentrations and metabolites were characterized using LC/MS and LC-MS/MS fragmentation patterns. Locomotor activity was monitored for 180240 min. Results Mephedrone plasma concentrations after i.v. administration fit a two-compartment model (α=10.23 h−1, β=1.86 h−1). After oral administration, peak mephedrone concentrations were achieved between 0.5 and 1 h and declined to undetectable levels at 9 h. The absolute bioavailability of mephedrone was about 10 % and the percentage of mephedrone protein binding was 21.59±3.67%. We have identified five phase I metabolites in rat blood after oral administration. The relationship between brain levels and free plasma concentration was 1.85±0.08. Mephedrone induced a dose-dependent increase in locomotor activity, which lasted up to 2 h. The pharmacokineticpharmacodynamic model successfully describes the relationship between mephedrone plasma concentrations and its psychostimulant effect. Conclusions We suggest a very important first-pass effect for mephedrone after oral administration and an easy access to the central nervous system. The model described might be useful in the estimation and prediction of the onset, magnitude,and time course of mephedrone pharmacodynamics as well as to design new animal models of mephedrone addiction and toxicity.

Role of P-glycoprotein in the uptake/efflux transport of oral vitamin K antagonists and rivaroxaban through the Caco-2 cell model.

Vitamin K antagonists (VKAs) are prescribed worldwide and remain the oral anticoagulant of choice. These drugs are characterized by a narrow therapeutic index and a large inter- and intra-individual variability. P-glycoprotein could contribute to this variability. The aim of this study was to investigate the involvement of P-gp in the transport of acenocoumarol, phenprocoumon and warfarin using an in vitro Caco-2 cell monolayer model. These results were compared with those obtained with rivaroxaban, a new oral anticoagulant known to be a P-gp substrate. The transport of these four drugs was assessed at pH conditions 6.8/7.4 in the presence or absence of the P-gp inhibitor cyclosporine A (10 μM) and the more potent and specific P-gp inhibitor valspodar (5 μM). Analytical quantification was performed by LC/MS. With an efflux ratio of 1.7 and a significant decrease in the efflux (Papp B-A), in the presence of P-gp inhibitors at a concentration of 50 μM, acenocoumarol can be considered as a weak P-gp substrate. Concerning phenprocoumon, the results suggest that this molecule is a poor P-gp substrate. The P-gp inhibitors did not affect significantly the transport of warfarin. The efflux of rivaroxaban was strongly inhibited by the two P-gp inhibitors. In conclusion, none of the three VKAs tested are strong P-gp substrates. However, acenocoumarol can be considered as a weak P-gp substrate and phenprocoumon as a poor P-gp substrate.

Comparative transcriptomics of early dipteran development

BACKGROUND: Modern sequencing technologies have massively increased the amount of data available for comparative genomics. Whole-transcriptome shotgun sequencing (RNA-seq) provides a powerful basis for comparative studies. In particular, this approach holds great promise for emerging model species in fields such as evolutionary developmental biology (evo-devo). RESULTS: We have sequenced early embryonic transcriptomes of two non-drosophilid dipteran species: the moth midge Clogmia albipunctata, and the scuttle fly Megaselia abdita. Our analysis includes a third, published, transcriptome for the hoverfly Episyrphus balteatus. These emerging models for comparative developmental studies close an important phylogenetic gap between Drosophila melanogaster and other insect model systems. In this paper, we provide a comparative analysis of early embryonic transcriptomes across species, and use our data for a phylogenomic re-evaluation of dipteran phylogenetic relationships. CONCLUSIONS: We show how comparative transcriptomics can be used to create useful resources for evo-devo, and to investigate phylogenetic relationships. Our results demonstrate that de novo assembly of short (Illumina) reads yields high-quality, high-coverage transcriptomic data sets. We use these data to investigate deep dipteran phylogenetic relationships. Our results, based on a concatenation of 160 orthologous genes, provide support for the traditional view of Clogmia being the sister group of Brachycera (Megaselia, Episyrphus, Drosophila), rather than that of Culicomorpha (which includes mosquitoes and blackflies).

Oral rivaroxaban for symptomatic venous thromboembolism.

BACKGROUND: Rivaroxaban, an oral factor Xa inhibitor, may provide a simple, fixed-dose regimen for treating acute deep-vein thrombosis (DVT) and for continued treatment, without the need for laboratory monitoring. METHODS: We conducted an open-label, randomized, event-driven, noninferiority study that compared oral rivaroxaban alone (15 mg twice daily for 3 weeks, followed by 20 mg once daily) with subcutaneous enoxaparin followed by a vitamin K antagonist (either warfarin or acenocoumarol) for 3, 6, or 12 months in patients with acute, symptomatic DVT. In parallel, we carried out a double-blind, randomized, event-driven superiority study that compared rivaroxaban alone (20 mg once daily) with placebo for an additional 6 or 12 months in patients who had completed 6 to 12 months of treatment for venous thromboembolism. The primary efficacy outcome for both studies was recurrent venous thromboembolism. The principal safety outcome was major bleeding or clinically relevant nonmajor bleeding in the initial-treatment study and major bleeding in the continued-treatment study. RESULTS: The study of rivaroxaban for acute DVT included 3449 patients: 1731 given rivaroxaban and 1718 given enoxaparin plus a vitamin K antagonist. Rivaroxaban had noninferior efficacy with respect to the primary outcome (36 events [2.1%], vs. 51 events with enoxaparin-vitamin K antagonist [3.0%]; hazard ratio, 0.68; 95% confidence interval [CI], 0.44 to 1.04; P<0.001). The principal safety outcome occurred in 8.1% of the patients in each group. In the continued-treatment study, which included 602 patients in the rivaroxaban group and 594 in the placebo group, rivaroxaban had superior efficacy (8 events [1.3%], vs. 42 with placebo [7.1%]; hazard ratio, 0.18; 95% CI, 0.09 to 0.39; P<0.001). Four patients in the rivaroxaban group had nonfatal major bleeding (0.7%), versus none in the placebo group (P=0.11). CONCLUSIONS: Rivaroxaban offers a simple, single-drug approach to the short-term and continued treatment of venous thrombosis that may improve the benefit-to-risk profile of anticoagulation. (Funded by Bayer Schering Pharma and Ortho-McNeil; ClinicalTrials.gov numbers, NCT00440193 and NCT00439725.).

Moniensiöinen muuntaja taajuudenmuuttajakäytössä

Suuritehoisissa pumppu- ja puhallinkäytöissä käytetään usein suurnopeusmoottoria,jota syötetään välijännitteellä. Suurjännitetaajuudenmuuttajat ovat kalliita, eikä niitä ole aina edes mahdollista valmistaa. Tutkimuksen kohteena on rinnakkaisilla pienjännitetaajuudenmuuttajilla toteutettu sähkökäyttö, jossa pienjännite nostetaan moniensiöisellä muuntajalla suurjännitteeksi (6,6 kV) ja syötetään edelleen kuormana olevalle suurnopeusmoottorille. Opinnäytetyössä tutkitaan moniensiöisen muuntajan syöttöä rinnakkaisilla taajuudenmuuttajilla sekä niiden aiheuttamia häiriöitä toisilleen. Työssä tutkitaan myös harmonisten yliaaltojen vaikutuksia muuntajan häviöihin ja magnetointiominaisuuksiin. Taajuudenmuuttajan lähtöjännite ja -virta suodatetaan sinisuotimella, jonka parametreja simuloidaan Simulink- ohjelmistolla. Tavoite on löytää optimaaliset parametrit taajuudenmuuttajanlähtösuotimelle käyrämuotojen ja suotimeen jäävän tehon suhteen. Työssä tarkasteltiin sinisuodinta, johon jää 3 prosenttia syöttöjännitteestä. LC-suodin kompensoi sähkökäytön loistehon lähes kokonaan, joten taajuudenmuuttajien antotehon kannalta suotimet ovat perusteltuja. Taajuudenmuuttajan näennäisteho putoaa 22 prosenttia, joten taajuudenmuuttajat voidaan vastaavasti mitoittaa pienemmiksi.

O USO DE CLORETO DE CÁLCIO E DA CAL PARA O TRATAMENTO PÓS-COLHEITA DE PODRIDÕES EM MAÇÃS

Foi desenvolvido um experimento para avaliar a influência da contaminação da água e a eficiência de produtos químicos na lavagem de maçãs cvs. Gala e Fuji sobre a ocorrência de podridão em frutos com ferimentos. O delineamento experimental foi inteiramente casualizado, com quatro repetições por cultivar e unidade experimental composta por 25 frutos. Os tratamentos foram: a) Testemunha (seca); b) Testemunha em água; c) Inoculação com água com esporos; d) Inoculação em água com esporos + 30 horas em temperatura ambiente; e) 30 horas em temperatura ambiente + inoculação em água com esporos; f) Cal Ca(OH)2 (1,5%); g) CaCl2 (1,5%). Inicialmente, todos os frutos sofreram quatro lesões de 0,2cm de diâmetro por 0,5cm de profundidade na região equatorial. Os frutos foram inoculados com uma solução de esporos de Penicillium sp. Após aplicação dos respectivos tratamentos, os frutos foram armazenados sob refrigeração a 0ºC para a 'Gala' e -0,5ºC para a 'Fuji'. As avaliações de incidência de podridão foram realizadas na abertura das câmaras (60 dias) e após 7 e 14 dias de exposição a 20ºC. Não houve ocorrência de podridão aos 60 dias para os frutos tratados com cal, não diferindo estatisticamente dos tratados com CaCl2. Aos sete e 14 dias, a cal mostrou-se mais eficiente que o CaCl2 na cv. Gala. Os frutos que ficaram 30 horas em temperatura ambiente antes de serem inoculados com uma solução de esporos, apresentaram menor incidência de podridão que os inoculados antes da exposição por 30 horas à temperatura ambiente, indicando que, após a inoculação, o fungo necessita de temperatura adequada para causar podridão.

Effect of water contamination on the shear bond strength of five orthodontic adhesives

Objectives: To evaluate the shear bond strength and site of failure of brackets bonded to dry and wet enamel. Study design: 50 teeth were divided into ten groups of 5 teeth each (10 surfaces). In half the groups enamel was kept dry before bonding, and in the other half distilled water was applied to wet the surface after etching. The following groups were established: 1)Acid/Transbond-XT (dry/wet) XT; 2) Transbond Plus Self Etching Primer (TSEP)/Transbond-XT paste (dry/wet); 3) Concise (dry), Transbond MIP/Concise (wet), 4) FujiOrtho-LC (dry/wet); 5) SmartBond (dry/wet). Brackets were bonded to both buccal and lingual surfaces. Specimens were stored in distilled water (24 hours at 37ºC) and thermocycled. Brackets were debonded using a Universal testing machine (cross-head speed 1 mm/min). Failure sites were classified using a stereomicroscope. Results: No significant differences in bond strength were detected between the adhesives under wet and dry conditions except for Smart- Bond, whose bond strength was significantly lower under dry conditions. For all the adhesives most bond failures were of mixed site location except for Smartbond, which failed at the adhesive-bracket interface. Conclusions: Under wet conditions the bonding capacity of the adhesives tested was similar than under dry conditions, with the exception of SmartBond which improved under wet conditions

EFFECTS OF THIDIAZURON (TDZ) ON FRUITING OF TEMPERATE TREE FRUITS

Thidiazuron (TDZ) is a phenylurea like citokinin on cell division fruit growth and fruit shape in some deciduous fruit trees. The effects of TDZ applied during flowering on apple cultivars 'Gala' and 'Fuji" were evaluated during seven growing seasons with annual applications on the same trees. The effects on pear and kiwi fruit trees were also evaluated. Every year, TDZ significantly increased fruit set and fruit weight on apple trees. The seven-year average of the fruit set from TDZ at 10 mg.L-1 was 112.7% while the control was only 51.3%. TDZ did not affect the number of clusters. The fruit weight increased 7.0% and 18.3% when the trees were sprayed with TDZ at 10 mg.L-1 and 5 mg.L-1, respectively. TDZ also increased fruit yield per tree by 28.7% and 41.8% for the 10 mg.L-1 and 5 mg.L-1 treatments, respectively. TDZ reduced the seed number per fruit and the calcium content in the flesh fruit, but increased the fruit firmness. The fruit set increased significantly on pear cultivar Packm's Triumph treated with TDZ, and reduced the seed numbers per fruit. TDZ applied at 12.5 mg.L-1 increased fruit weight by 47,4% on "Monty" kiwi.

Evaluation of a novel biomaterial in the suprachoroidal space of the rabbit eye.

PURPOSE: Drug delivery to treat diseases of the posterior segment of the eye, such as choroidal neovascularization and its complications, is hampered by poor intraocular penetration and rapid elimination of the drug from the eye. The purpose of this study was to investigate the feasibility and tolerance of suprachoroidal injections of poly(ortho ester) (POE), a bioerodible and biocompatible polymer, as a biomaterial potentially useful for development of sustained drug delivery systems. METHODS: After tunnelization of the sclera, different formulations based on POE were injected (100 microL) into the suprachoroidal space of pigmented rabbits and compared with 1% sodium hyaluronate. Follow-up consisted of fundus observations, echography, fluorescein angiography, and histologic analysis over 3 weeks. RESULTS: After injection, POE spread in the suprachoroidal space at the posterior pole. It was well tolerated and progressively disappeared from the site of injection without sequelae. No bleeding or retinal detachment occurred. Echographic pictures showed that the material was present in the suprachoroidal space for 3 weeks. Angiography revealed minor pigment irregularities at the site of injection, but no retinal edema or necrosis. Histology showed that POE was well tolerated in the choroid. CONCLUSIONS: POE suprachoroidal injections, an easy, controllable, and reproducible procedure, were well tolerated in the rabbit eye. POE appears to be a promising biomaterial to deliver drugs focally to the choroid and the retina.

Effect of aminoethoxyvinilglycine (AVG) on preharvest fruit drop and maturity of apples

Apple trees cultivars Gala and Fuji were sprayed four weeks before commercial harvest with aminoethoxyvinilglycine (AVG), at doses of 0, 125, or 250 mg L-1, and assessed for preharvest fruit drop, fruit growth, and maturation on tree. In 'Gala', 64 days after AVG spraying, fruit drop for control treatment was 85%, and AVG (at 125 and 250 mg L-1) reduced it to 10%. In 'Fuji', 64 days after AVG spraying, fruit drop for control was 6%, while treatments with AVG (at 125 and 250 mg L-1) increased fruit drop to 10%. AVG was a powerful retardant of fruit maturation for 'Gala' but not for 'Fuji'. In 'Gala', the most affected attribute was the skin background color, followed, in decreasing order, by soluble solids content, the starch index, skin red color, the flesh firmness, and titratable acidity. In 'Gala', only flesh firmness retention was improved by increasing AVG dose from 125 mg L-1 to 250 mg L-1. The AVG at 250 mg L-1 inhibited "Gala" late fruit growth but not 'Fuji'.

Efeito da tela antigranizo em algumas características dos frutos de macieira

A cobertura do pomar com telas de náilon é uma opção de combate ao granizo, principalmente em pequenas áreas. Durante cinco anos, avaliou-se o efeito do sombreamento provocado pela tela antigranizo, com níveis de sombreamento de 12%, 18% e 30%, sobre os fatores de produção na macieira, cvs. Gala e Fuji. A coloração vermelha dos frutos foi a variável mais afetada pelo sombreamento, reduzindo-se até 14,2% da coloração na cv. Gala e 7,2% na cv. Fuji. O sombreamento reduziu a incidência de "russeting" nos frutos, mas aumentou a incidência da mancha de glomerela. No último ano de avaliação, as plantas com cobertura de tela apresentaram uma redução de 19% no número de gemas com flor, quando comparadas com as plantas sem cobertura.

Effect of preharvest spraying with thidiazuron on fruit quality and maturity of apples

Apple trees, cultivars Gala and Fuji, were sprayed at full bloom with thidiazuron (TDZ) at the doses of 0, 5, 10, or 20 g (a.i.) ha-1 and fruit were assessed for quality and maturity. In both cultivars, the increase of TDZ dose had detrimental effects on fruit quality, causing a reduction of fruit red surface and an increase of percentage of fruit that was asymmetrical and with calyx-end rot. TDZ caused an increment of calyx-end aperture that might have increased calyx-end rot. TDZ increased density and the length : diameter (L : D) ratio of the fruit. The number of viable seeds was not affected by TDZ. TDZ resulted in a substantial delay of fruit maturity in 'Gala' but not in 'Fuji'. TDZ also reduced soluble solids content (SSC) in 'Gala' and reduced titratable acidity in both cultivars.

Efeito da intensidade do frio no tempo e percentagem de gemas brotadas em macieira

A macieira necessita de períodos de baixas temperaturas (4º a 10ºC) no outono e inverno, caso contrário a planta continuará em dormência ou apresentará uma brotação e floração irregular. Sem a ocorrência de baixas temperaturas no outono e inverno, as gemas da macieira continuam em dormência por um maior período. Para mensurar a quantidade de frio necessária para superar a dormência, o método utilizado foi o número de unidades de frio baseado no Modelo Carolina do Norte Modificado. Estacas de 20 a 25 cm de comprimento e estacas de nós isolados receberam zero, 530, 1060 e 1590 unidades de frio durante a dormência. As cultivares testadas foram 'Condessa', 'Baronesa', 'Daiane', 'Imperatriz', 'Gala' e 'Fuji'. As cultivares se diferenciaram quanto ao número de dias para a brotação das gemas, ocorrendo o menor tempo para a brotação quando receberam 1590 unidades de frio, para todas as cultivares, mostrando que há uma relação entre o tempo médio da brotação e a profundidade da dormência. A porcentagem de brotação das gemas foi maior quando as estacas foram submetidas a 1590 unidades de frio, sendo que a cultivar Condessa apresentou o maior percentual de brotação, confirmando as referências que indicam ser esta a cultivar de menor exigência em frio, dentre as estudadas.