Quantification of UV-B flux through time using UV-B-absorbing compounds contained in fossil Pinus sporopollenin

• UV-B radiation currently represents c. 1.5% of incoming solar radiation. However, significant changes are known to have occurred in the amount of incoming radiation both on recent and on geological timescales. Until now it has not been possible to reconstruct a detailed measure of UV-B radiation beyond c. 150 yr ago. • Here, we studied the suitability of fossil Pinus spp. pollen to record variations in UV-B flux through time. In view of the large size of the grain and its long fossil history, we hypothesized that this grain could provide a good proxy for recording past variations in UV-B flux. • Two key objectives were addressed: to determine whether there was, similar to other studied species, a clear relationship between UV-B-absorbing compounds in the sporopollenin of extant pollen and the magnitude of UV-B radiation to which it had been exposed; and to determine whether these compounds could be extracted from a small enough sample size of fossil pollen to make reconstruction of a continuous record through time a realistic prospect. • Preliminary results indicate the excellent potential of this species for providing a quantitative record of UV-B through time. Using this technique, we present the first record of UV-B flux during the last 9500 yr from a site near Bergen, Norway.

Adverbial quantification and perfective/imperfective interpretive nuances in L2 Portuguese

Slabakova (2006b) poses and directly addresses the question of whether or not there is a maturational effect (a critical/sensitive period) that affects the semantic component. She demonstrates that there is no empirical evidence suggesting that adults are unable to acquire phrasal semantic properties, even when the accessing of semantic universals is conditioned upon the acquisition of L2 morphosyntactic features (see Dekydtspotter and Sprouse 2001, Slabakova and Montrul 2003). In light of this, the authors test for interpretive properties associated with the aspectual projection higher (outer) AspP in advanced English learners of adult L2 Portuguese via their knowledge of [+/- accidental] related nuances in adverbially quantified preterit and imperfect sentences (Lenci and Bertinetto 2000; Menéndez-Benito 2002). In two experiments, the authors test for L2 knowledge of this [+/- accidental] distinction via semantic felicitousness judgments of adverbially quantified preterit and imperfect sentences depending on a supporting context as well as related restrictions on subject DP interpretations. Overall, the data show that advanced learners acquire this distinction. As the authors discuss, the present data support Full Access theories (White 1989, Schwartz and Sprouse 1996; Duffield and White 1999) and the No-Critical Period for semantics position (Slabakova 2006b), demonstrating that the syntax-semantics interface is not an inevitable locus for fossilization.

Identification and quantification of glucosinolate and flavonol compounds in rocket salad (Eruca sativa, Eruca vesicaria and Diplotaxis tenuifolia) by LC–MS: Highlighting the potential for improving nutritional value of rocket crops

Liquid Chromatography Mass Spectrometry (LC-MS) was used to obtain glucosinolate and flavonol content for 35 rocket accessions and commercial varieties. 13 glucosinolates and 11 flavonol compounds were identified. Semi-quantitative methods were used to estimate concentrations of both groups of compounds. Minor glucosinolate composition was found to be different between accessions; concentrations varied significantly. Flavonols showed differentiation between genera, with Diplotaxis accumulating quercetin glucosides and Eruca accumulating kaempferol glucosides. Several compounds were detected in each genus that have only previously been reported in the other. We highlight how knowledge of phytochemical content and concentration can be used to breed new, nutritionally superior varieties. We also demonstrate the effects of controlled environment conditions on the accumulations of glucosinolates and flavonols and explore the reasons for differences with previous studies. We stress the importance of consistent experimental design between research groups to effectively compare and contrast results.

Greenhouse gas emissions from waste management—assessment of quantification methods

Of the many sources of urban greenhouse gas (GHG) emissions, solid waste is the only one for which management decisions are undertaken primarily by municipal governments themselves and is hence often the largest component of cities’ corporate inventories. It is essential that decision-makers select an appropriate quantification methodology and have an appreciation of methodological strengths and shortcomings. This work compares four different waste emissions quantification methods, including Intergovernmental Panel on Climate Change (IPCC) 1996 guidelines, IPCC 2006 guidelines, U.S. Environmental Protection Agency (EPA) Waste Reduction Model (WARM), and the Federation of Canadian Municipalities- Partners for Climate Protection (FCM-PCP) quantification tool. Waste disposal data for the greater Toronto area (GTA) in 2005 are used for all methodologies; treatment options (including landfill, incineration, compost, and anaerobic digestion) are examined where available in methodologies. Landfill was shown to be the greatest source of GHG emissions, contributing more than three-quarters of total emissions associated with waste management. Results from the different landfill gas (LFG) quantification approaches ranged from an emissions source of 557 kt carbon dioxide equivalents (CO2e) (FCM-PCP) to a carbon sink of −53 kt CO2e (EPA WARM). Similar values were obtained between IPCC approaches. The IPCC 2006 method was found to be more appropriate for inventorying applications because it uses a waste-in-place (WIP) approach, rather than a methane commitment (MC) approach, despite perceived onerous data requirements for WIP. MC approaches were found to be useful from a planning standpoint; however, uncertainty associated with their projections of future parameter values limits their applicability for GHG inventorying. MC and WIP methods provided similar results in this case study; however, this is case specific because of similarity in assumptions of present and future landfill parameters and quantities of annual waste deposited in recent years being relatively consistent.

A high-density immunoblotting methodology for quantification of total protein levels and phosphorylation modifications

The components of many signaling pathways have been identified and there is now a need to conduct quantitative data-rich temporal experiments for systems biology and modeling approaches to better understand pathway dynamics and regulation. Here we present a modified Western blotting method that allows the rapid and reproducible quantification and analysis of hundreds of data points per day on proteins and their phosphorylation state at individual sites. The approach is of particular use where samples show a high degree of sample-to-sample variability such as primary cells from multiple donors. We present a case study on the analysis of >800 phosphorylation data points from three phosphorylation sites in three signaling proteins over multiple time points from platelets isolated from ten donors, demonstrating the technique's potential to determine kinetic and regulatory information from limited cell numbers and to investigate signaling variation within a population. We envisage the approach being of use in the analysis of many cellular processes such as signaling pathway dynamics to identify regulatory feedback loops and the investigation of potential drug/inhibitor responses, using primary cells and tissues, to generate information about how a cell's physiological state changes over time.

Quantification of major camel milk proteins by capillary electrophoresis

Proteins from dromedary camel milk (CM) produced in Europe were separated and quantified by capillary electrophoresis (CE). CE analysis showed that camel milk lacks b-lactoglobulin and consists of high concentration of a-lactalbumin (2.01 ± 0.02 mg mL-1), lactoferrin (1.74 ± 0.06 mg mL-1) and serum albumin (0.46 ± 0.01 mg mL-1 ). Among caseins, the concentration of b-casein (12.78 ± 0.92 mg mL-1) was found the highest followed by a-casein (2.89 ± 0.29 mg mL-1) while k-casein represented only minor amount (1.67 ± 0.01 mg mL-1). These results were in agreement with sodium dodecyl sulphatepolyacrylamide gel electrophoresis patterns. Overall, CE offers a quick and reliable method for the determination of major CM proteins, which may be responsible for the many nutritional and health properties of CM.

Listeria monocytogenes in ready-to-eat, sliced, cooked ham and salami products, marketed in the city of Sao Paulo, Brazil Occurrence, quantification, and serotyping

The preference for ready-to-eat sliced foods may pose an increased risk for food-borne diseases and a major concern is the presence of Listeria monocytogenes L monocytogenes was assessed in two types of products cooked ham and salami One hundred and thirty samples of each product were acquired in retail shops in the city of Sao Paulo and submitted to laboratory analysis The rate of positives was significantly higher in salami samples than in ham samples (62% and 0 8% respectively) L. monocytogenes counts in salami samples varied between <10 and 1900 colony-forming units per gram (CFU/g) The serotypes found in both products were as follows according to incidence 4b (37 5%) 1/2b (25%) 3b (25%) and 1/2c (12 5%) Based on the results of the present study the authors suggest that the risk of listeriosis resulting from the consumption of salami is higher than that associated with the consumption of cooked ham (C) 2010 Elsevier Ltd All rights reserved

Quantification of chlordesmethyldiazepam by liquid chromatography-tandem mass spectrometry: application to a cloxazolam bioequivalence study

A rapid, sensitive and specific LC-MS/MS method was developed and validated for quantifying chlordesmethyldiazepam (CDDZ or delorazepam), the active metabolite of cloxazolam, in human plasma. In the analytical assay, bromazepam (internal standard) and CDDZ were extracted using a liquid-liquid extraction (diethyl-ether/hexane, 80/20, v/v) procedure. The LC-MS/MS method on a RP-C18 column had an overall run time of 5.0 min and was linear (1/x weighted) over the range 0.5-50 ng/mL (R > 0.999). The between-run precision was 8.0% (1.5 ng/mL), 7.6% (9 ng/mL), 7.4% (40 ng/mL), and 10.9% at the low limit of quantification-LLOQ (0.500 ng/mL). The between-run accuracies were 0.1, -1.5, -2.7 and 8.7% for the above mentioned concentrations, respectively. All current bioanalytical method validation requirements (FDA and ANVISA) were achieved and it was applied to the bioequivalence study (Cloxazolam-test, Eurofarma Lab. Ltda and Olcadil (R)-reference, Novartis Biociencias S/A). The relative bioavailability between both formulations was assessed by calculating individual test/reference ratios for Cmax, AUClast and AUCO-inf. The pharmacokinetic profiles indicated bioequivalence since all ratios were as proposed by FDA and ANVISA. Copyright (C) 2009 John Wiley & Sons, Ltd.

Ciprofibrate quantification in human plasma by high-performance liquid chromatography coupled with electrospray tandem mass spectrometry for pharmacokinetic studies

A rapid, sensitive and specific method for quantifying ciprofibrate in human plasma using bezafibrate as the internal standard (IS) is described. The sample was acidified prior extraction with formic acid (88%). The analyte and the IS were extracted from plasma by liquid-liquid extraction using an organic solvent (diethyl ether/dichloromethane 70/30 (v/v)). The extracts were analyzed by high performance liquid chromatography coupled with electrospray tandem mass spectrometry (HPLC-MS/MS). Chromatography was performed using Genesis C18 4 mu m analytical column (4.6 x 150 mm i.d.) and a mobile phase consisting of acetonitrile/water (70/30, v/v) and 1 mM acetic acid. The method had a chromatographic run time of 3.4 min and a linear calibration curve over the range 0.1-60 mu g/mL (r > 0.99). The limit of quantification was 0.1 mu g/mL. The intra- and interday accuracy and precision values of the assay were less than 13.5%. The stability tests indicated no significant degradation. The recovery of ciprofibrate was 81.2%, 73.3% and 76.2% for the 0.3, 5.0 and 48.0 ng/mL standard concentrations, respectively. For ciprofibrate, the optimized parameters of the declustering potential, collision energy and collision exit potential were -51 V, -16 eV and -5 V, respectively. The method was also validated without the use of the internal standard. This HPLC-MS/MS procedure was used to assess the bioequivalence of two ciprofibrate 100 mg tablet formulations in healthy volunteers of both sexes. The following pharmacokinetic parameters were obtained from the ciprofibrate plasma concentration vs. time curves: AUC(last), AUC(0-168 h), C(max) and T(max). The geometric mean with corresponding 90% confidence interval (CI) for test/reference percent ratios were 93.80% (90% CI = 88.16-99.79%) for C(max), 98.31% (90% CI = 94.91-101.83%) for AUC(last) and 97.67% (90% CI = 94.45-101.01%) for AUC(0-168 h). Since the 90% Cl for AUC(last), AUC(0-168 h) and C(max) ratios were within the 80-125% interval proposed by the US FDA, it was concluded that ciprofibrate (Lipless (R) 100 mg tablet) formulation manufactured by Biolab Sanus Farmaceutica Ltda. is bioequivalent to the Oroxadin (R) (100 mg tablet) formulation for both the rate and the extent of absorption. (C) 2011 Published by Elsevier B.V.

Quantification of Porphyromonas gingivalis and fimA genotypes in smoker chronic periodontitis

Teixeira SRL, Mattarazo F, Feres M, Figueiredo LC, de Faveri M, Simionato MRL, Mayer MPA. Quantification of Porphyromonas gingivalis and fimA genotypes in smoker chronic periodontitis. J Clin Periodontol 2009; 36: 482-487. doi: 10.1111/j.1600-051X.2009.01411.x. Porphyromonas gingivalis fimA genotypes were associated with virulence factors in vitro, but little evidence of an association with disease severity were shown in humans. We aimed to correlate levels of P. gingivalis fimA genotypes II and IV and probing depth in smoker-chronic periodontitis subjects. One hundred and sixty eight subgingival samples of 20 smokers non-treated chronic periodontitis subjects obtained from sites with different probing depths [shallow (<= 3 mm), intermediate (4-6 mm), deep (>= 7 mm)] were analysed by real-time PCR for P. gingivalis and genotypes fimA II and IV. P. gingivalis and fimA IV were detected in all subjects, whereas fimA II was detected in 18 subjects (90%). One hundred and fifty two sites (90.5%) harboured P. gingivalis. Genotypes II and IV were detected in 28% and 69.6% of sites, respectively. The proportions of genotypes II and IV in relation to P. gingivalis levels were similar in shallow, intermediate and deep probing sites (2.4%, 4.6%, 1.4% for genotype II and 15.5%, 17.7%, 11.7% for genotype IV, respectively), indicating that other non-tested genotypes were more abundant. Increased levels of genotype IV were associated with increasing probing depth, but not of genotype II. The data suggested an association between P. gingivalis genotype fimA IV and disease severity in smoker-chronic periodontitis subjects.

A real-time polymerase chain reaction assay for the identification and quantification of American Leishmania species on the basis of glucose-6-phosphate dehydrogenase

A real-time polymerase chain reaction (PCR) test was developed on the basis of the Leishmania glucose-6-phosphate dehydrogenase locus that enables identification and quantification of parasites. Using two independent pairs of primers in SYBR-Green assays, the test identified etiologic agents of cutaneous leishmaniasis belonging to both subgenera, Leishmania (Viannia) and Leishmania (Leishmania) in the Americas. Furthermore, use of TaqMan probes enables distinction between L. (V.) braziliensis or L. (V.) peruviania from the other L. (Viannia) species. All assays were negative with DNA of related trypanosomatids, humans, and mice. The parasite burden was estimated by normalizing the number of organisms per total amount of DNA in the sample or per host glyceraldehyde-3-phosphate dehydrogenase copies. The real-time PCR assay for L. (Leishmania) subgenus showed a good linear correlation with quantification on the basis of a limiting dilution assay in experimentally infected mice. The test successfully identifies and quantifies Leishmania in human biopsy specimens and represents a new tool to study leishmaniasis.

Recurrence quantification analysis of electrostatic fluctuations in fusion plasmas

We have investigated plasma turbulence at the edge of a tokamak plasma using data from electrostatic potential fluctuations measured in the Brazilian tokamak TCABR. Recurrence quantification analysis has been used to provide diagnostics of the deterministic content of the series. We have focused our analysis on the radial dependence of potential fluctuations and their characterization by recurrence-based diagnostics. Our main result is that the deterministic content of the experimental signals is most pronounced at the external part of the plasma column just before the plasma radius. Since the chaoticity of the signals follows the same trend, we have concluded that the electrostatic plasma turbulence at the tokamak plasma edge can be partially explained by means of a deterministic nonlinear system. (C) 2007 Elsevier B.V. All rights reserved.