A rapid and simple uhplc-ms/ms method for quantification of plasma globotriaosylsphingosine (Lyso-gb3)

Fabry disease (FD) is a rare X-linked lysosomal storage disorder caused by α-galactosidase A gene (GLA) mutations, resulting in loss of activity of the lysosomal hydrolase, α-galactosidase A (α-Gal A). As a result, the main glycosphingolipid substrates, globotriaosylceramide (Gb3) and globotriaosylsphingosine (lyso-Gb3), accumulate in plasma, urine, and tissues. Here, we propose a simple, fast, and sensitive method for plasma quantification of lyso-Gb3, the most promising secondary screening target for FD. Assisted protein precipitation with methanol using Phree cartridges was performed as sample pre-treatment and plasma concentrations were measured using UHPLC-MS/MS operating in MRM positive electrospray ionization. Method validation provided ex-cellent results for the whole calibration range (0.25–100 ng/mL). Intra-assay and inter-assay accuracy and precision (CV%) were calculated as <10%. The method was successfully applied to 55 plasma samples obtained from 34 patients with FD, 5 individuals carrying non-relevant polymorphisms of the GLA gene, and 16 healthy controls. Plasma lyso-Gb3 concentrations were larger in both male and female FD groups compared to healthy subjects (p < 0.001). Normal levels of plasma lyso-Gb3 were observed for patients carrying non-relevant mutations of the GLA gene compared to the control group (p = 0.141). Dropping the lower limit of quantification (LLOQ) to 0.25 ng/mL allowed us to set the optimal plasma lyso-Gb3 cut-off value between FD patients and healthy controls at 0.6 ng/mL, with a sensitivity of 97.1%, specificity of 100%, and accuracy of 0.998 expressed by the area under the ROC curve (C.I. 0.992 to 1.000, p-value < 0.001). Based on the results obtained, this method can be a reliable tool for early phenotypic assignment, assessing diagnoses in patients with borderline GalA activity, and confirming non-relevant mutations of the GLA gene

Characterization of humic fractions in leachates from soil under organic and conventional management and their interactions with the root zone

Humic fractions were shown to be closely involved in gene expression and promotion of different PM H+-ATPase isoforms, as well as in lateral root development, indicating an enhanced nutrient absorption capacity of the plant root system. HPLC-SEC confirmed that water-soluble humic substances (WSHS) correspond to a subfraction of the fulvic fraction of humic substances. This was supported by E465/E665 ratios higher than 8.5. These ratios generally increased over the growing season in cultivated soils but showed significant differences between conventionally and organically managed bare soils. FTIR data and the analytical quantification of carboxyls confirmed relevant structural changes in bare soil under both organic and conventional farming management. Absorption intensities ratios at 1,590\u20131,570 cm-1 and 1,440\u20131,380 cm-1 showed the predominant aliphatic character of these molecules

Characterization of humic fractions in leachates from soil under organic and conventional management and their interactions with the root zone

Humic fractions were shown to be closely involved in gene expression and promotion of different PM H+-ATPase isoforms, as well as in lateral root development, indicating an enhanced nutrient absorption capacity of the plant root system. HPLC-SEC confirmed that water-soluble humic substances (WSHS) correspond to a subfraction of the fulvic fraction of humic substances. This was supported by E465/E665 ratios higher than 8.5. These ratios generally increased over the growing season in cultivated soils but showed significant differences between conventionally and organically managed bare soils. FTIR data and the analytical quantification of carboxyls confirmed relevant structural changes in bare soil under both organic and conventional farming management. Absorption intensities ratios at 1,590–1,570 cm-1 and 1,440–1,380 cm-1 showed the predominant aliphatic character of these molecules

The AMS-02 Time of Flight System. Final Design

The AMS-02 detector is a superconducting magnetic spectrometer that will operate on the International Space Station. The time of flight (TOF) system of AMS-02 is composed by four scintillator planes with 8, 8, 10, 8 counters each, read at both ends by a total of 144 phototubes. This paper describes the new design, the expected performances, and shows preliminary results of the ion beam test carried on at CERN on October 2002.Comment: 4 pages, 6 EPS figures. Proc. of the 28th ICRC (2003

Organic Amendments to Short Rotation Coppice (SRC) Plantation Affect Species Richness and Metal Accumulation of Spontaneously Growing Herbaceous Plants

Excess potentially toxic metals (PTMs) in soils require ad hoc approaches to salvage. Hence, this study explored the shoot accumulation of cadmium (Cd), lead (Pb), and zinc (Zn) by herbaceous plants growing under previously established Salix and Populus clones Short Rotation Coppice (SRC) with compost and sewage sludge applications in an abandoned metallurgical site, Podlesí, Czech Republic; PTM decontamination of soils. Soils within the SRC experimental site and outside considered as control were analyzed for their chemical properties by multi-analytical techniques. Shoots of spontaneously growing herbaceous plants under trees in the site and without trees in control were determined for pseudo-total Cd, Pb, and Zn contents. Moderately to slightly acid soils, high cation exchange capacity, and C/N ratio supported mineralization and relative mobility of total Cd (7.7–9.76), Pb (1541–1929), and Zn (245–320 mg kg−1) in soils. Although soil amendments improved chemical properties, compost application supported higher species richness than sewage sludge. Over 95% of plants accumulated Cd and Zn above the WHO threshold and green fodder in the Czech Republic, with 36% Pb above the regional limit (40 mg kg−1). Approximately 100, 50, and 6% of herbaceous species had Cd, Pb, and Zn accumulation, respectively, higher than published average upper limits in plants (0.2 Cd, 10 Pb, and 150 Zn mg kg−1). Dicots recorded higher Cd content, Tenacetum vulgare (L.), Hypericum maculatum (Crantz), and Cirsium arvense (L.); Stachys palustris (L.), Lamium perpereum (L.), and Campanula patula (L.) for Pb; Glechoma hederaceae (L.), C. patula, and C. arvense for Zn in all treatments. Appropriate soil amelioration of SRC-supported PTM mobility and excess herbaceous species shoot accumulation, growth, and richness

On the electromagnetic energy resolution of Cherenkov-fiber calorimeters

Electromagnetic calorimeters which sample the Cherenkov radiation of shower particles in optical fibers operate in a markedly different manner from calorimeters which rely on the dE/dx of shower particles. The well-understood physics of electromagnetic shower development is applied to the case of Cherenkov-fiber calorimetry (also known as quartz fiber calorimetry) and the results of systematically performed studies are considered in detail to derive an understanding of the critical parameters involved in energy measurement using such calorimeters. A quantitative parameterization of Cherenkov-fiber calorimetry electromagnetic energy resolution is proposed and compared with existing experimental results

The TOF counters of the AMS-02 experiment: space qualification tests and beam test results

The scintillator counters of the TOF system of AMS-02 is beeing constructed to match the needs of the AMS-02 experiment that is armed by a high aperture superconducting dipole magnet. The goals of the TOF-02 hodoscopes actually are: to give the fast trigger to the all sub-detectors of AMS-02; to measure the particle velocity ensuring a 1 × 10 9 albedo rejection; to measure the absolute charge by particle energy loss, up to at least Z = 20 . In spring of 2005 all the TOF counter planes will be assembled and the space qualification tests will be performed. A description of the first test results and of the TOF performances will be given

Study of single muons with the Large Volume Detector at Gran Sasso Laboratory

The present study is based on the sample of about 3 mln single muons observed by LVD at underground Gran Sasso Laboratory during 36500 live hours from June 1992 to February 1998. We have measured the muon intensity at slant depths from 3 km w.e. to 20 km w.e. Most events are high energy downward muons produced by meson decay in the atmosphere. The analysis of these muons has revealed the power index of pion and kaon spectrum: 2.76 \pm 0.05. The reminders are horizontal muons produced by the neutrino interactions in the rock surrounding LVD. The value of this flux is obtained. The results are compared with Monte Carlo simulations and the world data.Comment: 13 pages, 2 figures, accepted for publication in "Physics of Atomic Nuclei