Orthogonal organic and aqueous-based washes of tissue sections to enhance protein sensitivity by MALDI imaging mass spectrometry.

Imaging mass spectrometry (IMS) is an emergent and innovative approach for measuring the composition, abundance and regioselectivity of molecules within an investigated area of fixed dimension. Although providing unprecedented molecular information compared with conventional MS techniques, enhancement of protein signature by IMS is still necessary and challenging. This paper demonstrates the combination of conventional organic washes with an optimized aqueous-based buffer for tissue section preparation before matrix-assisted laser desorption/ionization (MALDI) IMS of proteins. Based on a 500 mM ammonium formate in water-acetonitrile (9:1; v/v, 0.1% trifluororacetic acid, 0.1% Triton) solution, this buffer wash has shown to significantly enhance protein signature by profiling and IMS (~fourfold) when used after organic washes (70% EtOH followed by 90% EtOH), improving the quality and number of ion images obtained from mouse kidney and a 14-day mouse fetus whole-body tissue sections, while maintaining a similar reproducibility with conventional tissue rinsing. Even if some protein losses were observed, the data mining has demonstrated that it was primarily low abundant signals and that the number of new peaks found is greater with the described procedure. The proposed buffer has thus demonstrated to be of high efficiency for tissue section preparation providing novel and complementary information for direct on-tissue MALDI analysis compared with solely conventional organic rinsing.

Effect of mannitol and hypertonic saline on cerebral oxygenation in patients with severe traumatic brain injury and refractory intracranial hypertension.

BACKGROUND: The impact of osmotic therapies on brain oxygen has not been extensively studied in humans. We examined the effects on brain tissue oxygen tension (PbtO(2)) of mannitol and hypertonic saline (HTS) in patients with severe traumatic brain injury (TBI) and refractory intracranial hypertension. METHODS: 12 consecutive patients with severe TBI who underwent intracranial pressure (ICP) and PbtO(2) monitoring were studied. Patients were treated with mannitol (25%, 0.75 g/kg) for episodes of elevated ICP (>20 mm Hg) or HTS (7.5%, 250 ml) if ICP was not controlled with mannitol. PbtO(2), ICP, mean arterial pressure, cerebral perfusion pressure (CPP), central venous pressure and cardiac output were monitored continuously. RESULTS: 42 episodes of intracranial hypertension, treated with mannitol (n = 28 boluses) or HTS (n = 14 boluses), were analysed. HTS treatment was associated with an increase in PbtO(2) (from baseline 28.3 (13.8) mm Hg to 34.9 (18.2) mm Hg at 30 min, 37.0 (17.6) mm Hg at 60 min and 41.4 (17.7) mm Hg at 120 min; all p<0.01) while mannitol did not affect PbtO(2) (baseline 30.4 (11.4) vs 28.7 (13.5) vs 28.4 (10.6) vs 27.5 (9.9) mm Hg; all p>0.1). Compared with mannitol, HTS was associated with lower ICP and higher CPP and cardiac output. CONCLUSIONS: In patients with severe TBI and elevated ICP refractory to previous mannitol treatment, 7.5% hypertonic saline administered as second tier therapy is associated with a significant increase in brain oxygenation, and improved cerebral and systemic haemodynamics.

The profiling of MDMA tablets: a study of the combination of physical characteristics and organic impurities as sources of information

The profiling of MDMA tablets can be carried out using different sets of characteristics. The first type of measurements performed on MDMA tablets are physical characteristics (i.e. post-tabletting characteristics). They yield preliminary profiling data that may be valuable in a first stage for investigation purposes. However organic impurities (i.e. pre-tabletting characteristics) are generally considered to bring more reliable information, particularly for presentation of evidence in court. This work aimed therefore at evaluating the added value of combining pre-tabletting characteristics and post-tabletting characteristics of seized MDMA tablets. In approximately half of the investigated cases, the post-tabletting links were confirmed with organic impurities analyses. In the remaining cases, post-tabletting batches (post-TBs) were divided in several pre-tabletting batches (pre-TBs), thus supporting the hypothesis that several production batches of MDMA powder (pre-TBs) were used to produce one single post-TB (i.e. tablets having the same shape, diameter, thickness, weight and score; but different organic impurities composition). In view of the obtained results, the hypotheses were discussed through illustrating examples. In conclusion, both sets of characteristics were found relevant alone and combined together. They actually provide distinct information about MDMA illicit production and trafficking.

Binding of ²³⁹Pu and ⁹⁰Sr to organic colloids in soil solutions: evidence from a field experiment.

Colloidal transport has been shown to enhance the migration of plutonium in groundwater downstream from contaminated sites, but little is known about the adsorption of ⁹⁰Sr and plutonium onto colloids in the soil solution of natural soils. We sampled soil solutions using suction cups, and separated colloids using ultrafiltration to determine the distribution of ²³⁹Pu and ⁹⁰Sr between the truly dissolved fraction and the colloidal fraction of the solutions of three Alpine soils contaminated only by global fallout from the nuclear weapon tests. Plutonium was essentially found in the colloidal fraction (>80%) and probably associated with organic matter. A significant amount of colloidal ⁹⁰Sr was detected in organic-rich soil solutions. Our results suggest that binding to organic colloids in the soil solutions plays a key role with respect to the mobility of plutonium in natural alpine soils and, to a lesser extent, to the mobility of ⁹⁰Sr.

Contrast ultrasound: a simple-to-use phase-shifting medium offers saline infusion sonography-like images.

OBJECTIVE: To test the ability of a novel phase-shifting medium (PSM) to provide sustained distension of the uterine cavity and produce saline infusion sonography (SIS)-like images in a simplified contrast ultrasound procedure. DESIGN: Prospective pilot feasibility trial of a new diagnostic procedure, contrast ultrasound. SETTING: Clinical reproductive endocrine and infertility unit of regional teaching hospital. PATIENT(S): Twenty-six asymptomatic infertile women (group I) and 27 women presenting with dysfunctional uterine bleeding (DUB) who were scheduled for exploratory surgery (group II). INTERVENTION(S): All women who were temporarily on oral contraceptive first had a regular pelvic ultrasound followed by the intrauterine instillation of up to 3 mL PSM, using a regular insemination catheter, after which all instruments were removed and a regular ultrasound was performed again. RESULT(S): In all 53 women, intrauterine instillation of 1-3 mL PSM resulted in a 3-7 mm uterine distension, sufficient to produce SIS-like images of the uterine cavity that lasted 7-10 min. Contrast ultrasound revealed an endometrial polyp in 3 asymptomatic women of group I. In group II. 12 of 14 women (86%) whose vaginal ultrasound were positive or dubious had positive findings with contrast ultrasound; 9 of 12 patients whose vaginal ultrasounds were negative also had positive contrast ultrasound findings. All the positive and negative findings of contrast ultrasound made in group II were confirmed anatomically (sensitivity and specificity of 100%), whereas the correlation for standard vaginal ultrasound was markedly lower at 57.1% and 85.7%, respectively. Most patients (46 of 53) reported no discomfort during or after the procedure, and 7 women described the procedure as mildly uncomfortable. CONCLUSION(S): Contrast ultrasound, a novel simple diagnostic procedure conducted after intrauterine instillation of 1-3 mL PSM using a simple plastic catheter, delivered SIS-quality images in asymptomatic (group I) and symptomatic (group II) patients while retaining the simplicity of standard ultrasound. We therefore foresee broad application of contrast ultrasound for sensitive and specific assessment for uterine pathologies in the physician's office.

Characterization of a protein of the plastid inner envelope having homology to animal inorganic phosphate, chloride and organic-anion transporters.

A protein from Arabidopsis thaliana (L.) Heynh. showing homology to animal proteins of the NaPi-1 family, involved in the transport of inorganic phosphate, chloride, glutamate and sialic acid, has been characterized. This protein, named ANTR2 (for anion transporters) was shown by chloroplast subfractionation to be localized to the plastid inner envelope in both A. thaliana and Spinacia oleracea (L.). Immunolocalization revealed that ANTR2 was expressed in the leaf mesophyll cells as well as in the developing embryo at the upturned-U stage. Five additional homologues of ANTR2 are found in the Arabidopsis genome, of which one was shown by green fluorescent protein (GFP) fusion to be also located in the chloroplast. All ANTR proteins share homology to the animal NaPi-1 family, as well as to other organic-anion transporters that are members of the Anion:Cation Symporter (ACS) family, and share the main features of transporters from this family, including the presence of 12 putative transmembrane domains and of a 7-amino acid motif in the fourth putative transmembrane domain. ANTR2 thus represent a novel protein of the plastid inner envelope that is likely to be involved in anion transport.

Chemical analyses of organic residues in archaeological pottery from Arbon Bleiche 3, Switzerland - evidence for dairying in the late Neolithic

Fatty acids distribution and stable isotope ratios (bulk delta(13)C. delta(15)N and delta(13)C of individual fatty acids) of organic residues from 30 potsherds have been used to get further insights into the diet at the Late Neolithic (3384-3370 BC) site of Arbon Bleiche 3. Switzerland. The results are compared with modern equivalents of animal and vegetable fats, which may have been consumed ill a mixed ecology community having agrarian, breeding, shepherd, gathering, hunting, and fishing activities. The used combined chemical and isotopic approach provides valuable information to complement archaeological indirect evidence about the dietary trends obtained from the analysis of faunal and plant remains. The small variations of the delta(13)C and delta(15)N values within the range expected for degraded animal and plant tissues, is consistent with the archaeological evidence of animals, whose subsistence was mainly based on C(3) plants. The overall fatty acid composition and the stable carbon isotopic compositions of palmitic, stearic and oleic acids of the organic residues indicate that the studied Arbon Bleiche 3 sherds contain fat residues of plant and animal origin, most likely ruminant (bovine and ovine). In several vessels the presence of milk residues provides direct evidence for dairying during the late Neolithic in central Europe. (c) 2005 Elsevier Ltd. All rights reserved.

Comparison of antibiotic drops placed in the conjunctival cul-de-sac to antibiotic ointment applied to the lid margin in reduction of bacterial colonization on the lid margin.

PURPOSE: To compare the efficacy of antibiotic drops placed in the conjunctival cul-de-sac to antibiotic ointment applied to the lid margin in reduction of bacterial colonization on the lid margin. METHODS: A randomized, prospective, single-masked study was conducted on 19 patients with culture-proven colonization of bacteria on the lid margins. Ophthalmic eligibility criteria included the presence of > or =50 colony-forming units/mL (CFU/mL) of bacteria on both right and left lids. Each patient received one drop of ofloxacin in one eye every night for one week, followed by one drop once a week for one month. In the same manner, each patient received bacitracin ointment (erythromycin or gentamicin ointment if lid margin bacteria were resistant to bacitracin) to the lid margin of the fellow eye. Quantitative lid cultures were taken at initial visit, one week, one month, and two months. Fifteen volunteers (30 lids) served as controls. Lid cultures were taken at initial visit, one week, and one month. RESULTS: Both antibiotic drop and ointment reduced average bacterial CFU/mL at one week and one month. Average bacterial CFU/mL reestablished to baseline values at two months. There was no statistically significant difference between antibiotic drop and ointment in reducing bacterial colonization on the lid margin. CONCLUSION: Antibiotic drops placed in the conjunctival cul-de-sac appear to be as effective as ointment applied to the lid margins in reducing bacterial colonization in patients with > or =50 CFU/mL of bacteria on the lid margins.

Late Pliensbachian-Early Toarcian (Early Jurassic) environmental changes in an epicontinental basin of NW Europe (Causses area, central France): A micropaleontological and geochemical approach

We present an integrated work based on calcareous nannofossil and benthic foraminiferal assemblages, and geochemical analyses of two Upper Pliensbachian-Lower Toarcian sections located in the central-South France. The studied sections, Tournadous and Saint-Paul-des-Fonts, represent the proximal and the distal part, respectively, of the Jurassic Causses Basin, one of the small, partly enclosed basins belonging to the epicontinental shelf of the NW Tethys. At the transition from Late Pliensbachian to Early Toarcian, the Causses Basin recorded an emersion in response to the global sea-level fall. Our data indicate severe environmental conditions of marine waters, including salinity decrease and anoxia development, occurring in the Early Toarcian. The acme of this deterioration coincides with the Early Toarcian Anoxic Event (T-OAE) but, due to the restricted nature of the basin. anoxia persisted until the end of the Early Toarcian. mainly in the deeper parts of the basin. The micronutrients and organic organic-matter fluxes were probably high during the entire studied time interval, as shown by nannofossil and foraminiferal assemblages. However, nannoplankton production drastically decreased during the T-OAE, as demonstrated by very low nannofossil fluxes, and only taxa tolerant to low-saline surface waters could thrive. At the same time, benthic foraminifers temporarily disappeared in response to sea-bottom anoxia. Our study demonstrates that environmental changes related to the T-OAE are well-recorded even in small, partly enclosed basins of NW Europe, like the Causses Basin. Within this area, the effects of global changes. like sea sea-level and temperature fluctuations, are modulated by local conditions mainly controlled by the morphology of the basin. (C) 2008 Elsevier B.V. All rights reserved.

δ 15N measurement of organic and inorganic substances by EA-IRMS: a speciation-dependent procedure

Little attention has been paid so far to the influence of the chemical nature of the substance when measuring δ 15N by elemental analysis (EA)-isotope ratio mass spectrometry (IRMS). Although the bulk nitrogen isotope analysis of organic material is not to be questioned, literature from different disciplines using IRMS provides hints that the quantitative conversion of nitrate into nitrogen presents difficulties. We observed abnormal series of δ 15N values of laboratory standards and nitrates. These unexpected results were shown to be related to the tailing of the nitrogen peak of nitrate-containing compounds. A series of experiments were set up to investigate the cause of this phenomenon, using ammonium nitrate (NH4NO3) and potassium nitrate (KNO3) samples, two organic laboratory standards as well as the international secondary reference materials IAEA-N1, IAEA-N2-two ammonium sulphates [(NH4)2SO4]-and IAEA-NO-3, a potassium nitrate. In experiment 1, we used graphite and vanadium pentoxide (V2O5) as additives to observe if they could enhance the decomposition (combustion) of nitrates. In experiment 2, we tested another elemental analyser configuration including an additional section of reduced copper in order to see whether or not the tailing could originate from an incomplete reduction process. Finally, we modified several parameters of the method and observed their influence on the peak shape, δ 15N value and nitrogen content in weight percent of nitrogen of the target substances. We found the best results using mere thermal decomposition in helium, under exclusion of any oxygen. We show that the analytical procedure used for organic samples should not be used for nitrates because of their different chemical nature. We present the best performance given one set of sample introduction parameters for the analysis of nitrates, as well as for the ammonium sulphate IAEA-N1 and IAEA-N2 reference materials. We discuss these results considering the thermochemistry of the substances and the analytical technique itself. The results emphasise the difference in chemical nature of inorganic and organic samples, which necessarily involves distinct thermochemistry when analysed by EA-IRMS. Therefore, they should not be processed using the same analytical procedure. This clearly impacts on the way international secondary reference materials should be used for the calibration of organic laboratory standards.

Mutism and Amnesia following High-Voltage Electrical Injury: Psychogenic Symptomatology Triggered by Organic Dysfunction?

Background: Mutism and dense retrograde amnesia are found both in organic and dissociative contexts. Moreover, dissociative symptoms may be modulated by right prefrontal activity. A single case, M.R., developed left hemiparesis, mutism and retrograde amnesia after a high-voltage electric shock without evidence of lasting brain lesions. M.R. suddenly recovered from his mutism following a mild brain trauma 2 years later. Methods: M.R.'s neuropsychological pattern and anatomoclinical correlations were studied through (i) language and memory assessment to characterize his deficits, (ii) functional neuroimaging during a standard language paradigm, and (iii) assessment of frontal and left insular connectivity through diffusion tractography imaging and transcranial magnetic stimulation. A control evaluation was repeated after recovery. Findings: M.R. recovered from the left hemiparesis within 90 days of the accident, which indicated a transient right brain impairment. One year later, neurobehavioral, language and memory evaluations strongly suggested a dissociative component in the mutism and retrograde amnesia. Investigations (including MRI, fMRI, diffusion tensor imaging, EEG and r-TMS) were normal. Twenty-seven months after the electrical injury, M.R. had a very mild head injury which was followed by a rapid recovery of speech. However, the retrograde amnesia persisted. Discussion: This case indicates an interaction of both organic and dissociative mechanisms in order to explain the patient's symptoms. The study also illustrates dissociation in the time course of the two different dissociative symptoms in the same patient.