Reference standards Research Papers - Academia.edu (original) (raw)

To provide an overview of changes in nursing education in the European Union (EU) within the framework of the Bologna Declaration, signed in 1999 by the European ministers of education, and to describe specific efforts and changes in... more

To provide an overview of changes in nursing education in the European Union (EU) within the framework of the Bologna Declaration, signed in 1999 by the European ministers of education, and to describe specific efforts and changes in Spain. Organizing Constructs: (a) the Bologna Declaration process, its recent reforms in all disciplines including nursing, and perspectives on future developments; (b) the Tuning Project, designed and carried out by a group of European universities to meet the challenges posed by the Bologna Declaration; and (c) efforts in a group of Spanish universities to promote higher education in nursing. Findings and Conclusions: These changes promote enhanced academic recognition, professionalism in nursing education, and graduates' competencies in practice in most European countries by specifying the undergraduate nursing degree as the minimal entrance level for practice and master's and doctoral programs for further career development.

Introduction-Ilex paraguariensis A. St. Hil. (mate) is known in several South American countries because of the use of its leaves in stimulant herbal beverages. High saponin contents were reported in mate leaves and unripe fruits that... more

Introduction-Ilex paraguariensis A. St. Hil. (mate) is known in several South American countries because of the use of its leaves in stimulant herbal beverages. High saponin contents were reported in mate leaves and unripe fruits that possess a dissimilar composition. Two LC-UV methods previously reported for mate saponins assay focused on mate leaves and the quantification of the less polar saponin fraction in mate fruits. Objective-To develop and validate a LC-UV method to assay the total content of saponins in unripe mate fruits and characterise the chemical structure of triterpenic saponins by UPLC/Q-TOF-MS. Methodology-From unripe fruits of mate a crude ethanolic extract was prepared (EX40) and the mate saponin fraction (MSF) purified by solid phase extraction. The LC-UV method was validated using ilexoside II as external standard. UPLC/Q-TOF-MS was adjusted from the LC-UV method to obtain the fragmentation patterns of the main saponins present in unripe fruits. Results-Both LC-UV and UPLC/Q-TOF-MS methods indicate a wide range of Ilex saponins polarity. The ilexoside II and total saponin content of EX40 were 8.20% (w/w) and 47.60% (w/w), respectively. The total saponin content in unripe fruits was 7.28% (w/w). The saponins present in MSF characterised by UPLC/Q-TOF-MS are derived mainly from ursolic/oleanolic, acetyl ursolic or pomolic acid. Conclusion-The validated LC-UV method was shown to be linear, precise, accurate and to cover several saponins previously isolated from Ilex species and could be applied for the quality control of unripe fruit saponins.

We studied a series of test conditions in a microtiter system to define the optimal method for determining the susceptibility of Cryptococcus neoformans to antifungal agents. Twenty-one isolates of C. neoformans were grown for 24 or 48 h... more

We studied a series of test conditions in a microtiter system to define the optimal method for determining the susceptibility of Cryptococcus neoformans to antifungal agents. Twenty-one isolates of C. neoformans were grown for 24 or 48 h in four chemically defined media: yeast nitrogen base (BYNB 7); RPMI 1640; synthetic amino acid medium--fungal (SAAMF), buffered at pH 7.0 to select the medium that best supported growth of this fastidious yeast; and yeast nitrogen base, pH 5.4 (YNB 5.4). Maximum growth of C. neoformans, at 35 degrees C, was obtained in YNB 5.4, with the next highest growth levels in BYNB 7, SAAMF, and RPMI. Growth at 24 h was uniformly poor in all media and lacked reproducibility. In contrast, incubation for 48 h gave adequate growth with low standard deviations, and 48 h was selected as the optimal incubation period for this study. Comparison of the relationship between growth kinetics and initial inoculum size for eight cryptococcal isolates showed that 10(4) cel...

We measured apolipoproteins (apo) A-I and B by rate immunonephelometry (rate INA) during Phase 1 of the National Health and Nutrition Examination Survey (NHANES) III. We also made the measurements by radial immunodiffusion (RID) in a 20%... more

We measured apolipoproteins (apo) A-I and B by rate immunonephelometry (rate INA) during Phase 1 of the National Health and Nutrition Examination Survey (NHANES) III. We also made the measurements by radial immunodiffusion (RID) in a 20% subset of the samples. Aliquots of this subset were also analyzed in the Northwest Lipid Research Laboratories by fixed-time INA calibrated to the World Health Organization (WHO)-International Federation of Clinical Chemistry (IFCC) First International Reference Materials for Apolipoproteins A-I and B. The CVs for the rate INA and RID measurements were: apoA-I, 4.5-7.7% and 2.5-7.6%, respectively; apoB, 2.3-5.3% and 2.3-6.4%, respectively. In NHANES III, rate INA values (x) can be transformed to WHO-IFCC Reference Material-based values (y) as follows: for apoA-I, y = 0.87x + 251.8 mg/L (r = 0.93, SEslope = 0.13, SEintercept = 17, n = 708); for apoB (mg/L), y = 1.068x + 112.8 mg/L (r = 0.98, SEslope = 0.08, SEintercept = 7, n = 646).

The derivative from sucrose was prepared by substituting hydroxyl groups with chlorine. Identification of the modified sucrose was performed using the following analysis: Thin Layer chromatography (TLC), high performance liquid... more

The derivative from sucrose was prepared by substituting hydroxyl groups with chlorine. Identification of the modified sucrose was performed using the following analysis: Thin Layer chromatography (TLC), high performance liquid chromatography (HPLC), H-NMR, mass spectroscopy (MS), Inferred 1 spectroscopy (IR), melting point, OH groups. Moreover, some physical properties including: melting point, viscosity, surface tension and solubility.

A sensitive, specific and high throughput bioanalytical method using automated sample processing via 96-well plate liquid-liquid extraction and ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) has been... more

A sensitive, specific and high throughput bioanalytical method using automated sample processing via 96-well plate liquid-liquid extraction and ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) has been developed for the determination of methoxsalen in human plasma. Plasma samples with ketoconazole as internal standard (IS) were prepared by employing 0.2 mL human plasma in ethyl acetate:dichloromethane (80:20, v/v). The chromatographic separation was achieved on a Waters Acquity UPLC BEH C18 column using isocratic mobile phase, consisting of 10 mM ammonium formate and acetonitrile (60:40, v/v), at a flow rate of 0.5 mL/min. The linear dynamic range was established over the concentration range 1.1-213.1 ng/mL for methoxsalen. The method was rugged and rapid with a total run time of 1.5 min. It was successfully applied to a pivotal bioequivalence study in 12 healthy human subjects after oral administration of 10 mg extended release methoxsalen formulation under fasting condition.

Background: Different laboratory assays are used to detect and measure specific IgE antibodies. No standard exists to assess their analytic performance. Objective: We sought to analyze reported specific IgE results from different... more

Background: Different laboratory assays are used to detect and measure specific IgE antibodies. No standard exists to assess their analytic performance. Objective: We sought to analyze reported specific IgE results from different laboratories on the same serum samples for their accuracy and precision. Methods: Blinded serum samples (26) containing variable levels of specific IgE to 17 common aeroallergens were sent on 3 different occasions through normal channels to 6 laboratories that used 5 different test procedures. Six samples were presented as a dilution series. Laboratory-assay performance was assessed by analyzing the reported results (n = 12,708) by using ordinary least squares regression with slope coefficients, the t statistic, SEs, confidence intervals, and R 2 values. These were compared with a theoretic ideal assay as the reference. Results: Analysis revealed that one system used in two different laboratories performed nearly as well as the ideal standard, with an overall average slope (0.97; range, 0.91-1.01), SE (0.05; range, 0.02-0.16), R 2 value (93%; range, 0.64-0.99), and coefficient of variation (10.3%; range, 6%-14%). Extensive variability was observed in the other 4 laboratory-assay systems with respect to overall average slope (0.76; range, 0.11-1.24), SE (0.19; range, 0.03-0.95), R 2 value (53%; range, 0.00-0.98), and coefficient of variation (19%; range, 5%-49%). For some specific allergens, some laboratories-assays were not able to detect serial dilutions of the same sample. Conclusions: One commercial system used in two different laboratories performed nearly as well as the ideal standard. Four of the laboratories-assays for specific IgE antibodies demonstrated substandard overall performance with multiple instances of poor precision and accuracy, particularly for certain allergens, such as weeds and molds.

Objective: To assess the relationship of air pollution and hematologic parameters in a population-based sample of children and adolescents. Methods: This cross-sectional study was conducted in 2009-2010 among school students randomly... more

Objective: To assess the relationship of air pollution and hematologic parameters in a population-based sample of children and adolescents. Methods: This cross-sectional study was conducted in 2009-2010 among school students randomly selected from different areas of Isfahan city, the second largest and most air-polluted city in Iran. The association of air pollutants levels with hemoglobin, platelets, red and white blood cells (RBC and WBC, respectively) levels was determined by multiple linear and logistic regression analyses, after adjustment for age, gender, anthropometric measures, meteorological factors, and dietary and physical activity habits. Results: The study participants consisted of 134 students (48.5% boys) with a mean age of 13.10±2.21 years. While the mean Pollutant Standards Index (PSI) was at moderate level, the mean particulate matter ≤ 10 µm (PM 10) was more than twice the normal level. Multiple linear regression analysis showed that PSI and most air pollutants, notably PM 10 , had significant negative relationship with hemoglobin and RBC count, and positive significant relationship with WBC and platelet counts. The odds ratio of elevated WBC increased as the quartiles of PM 10 , ozone and PSI increased, however these associations reached to significant level only in the highest quartile of PM 10 and PSI. The corresponding figures for hemoglobin and RBC were in opposite direction. Conclusions: The association of air pollutants with hematologic parameters and a possible pro-inflammatory state is highlighted. The presence of these associations with PM 10 in a moderate mean PSI level underscores the necessity to reexamine environmental health policies for the pediatric age group.

Purpose: To assess the reliability of cone beam computed tomography (CBCT) voxel gray value measurements using Hounsfield units (HU) derived from multislice computed tomography (MSCT) as a clinical reference (gold standard). Materials and... more

Purpose: To assess the reliability of cone beam computed tomography (CBCT) voxel gray value measurements using Hounsfield units (HU) derived from multislice computed tomography (MSCT) as a clinical reference (gold standard). Materials and Methods: Ten partially edentulous human mandibular cadavers were scanned by two types of computed tomography (CT) modalities: multislice CT and cone beam CT. On MSCT scans, eight regions of interest (ROI) designating the site for preoperative implant placement were selected in each mandible. The datasets from both CT systems were matched using a three-dimensional (3D) registration algorithm. The mean voxel gray values of the region around the implant sites were compared between MSCT and CBCT. Results: Significant differences between the mean gray values obtained by CBCT and HU by MSCT were found. In all the selected ROIs, CBCT showed higher mean values than MSCT. A strong correlation (R = 0.968) between mean voxel gray values of CBCT and mean HU of...

Background: Conventional methods for isolation and antibiotic susceptibility reports of bacterial isolates from positive blood culture bottles takes at least 48 hours. Direct susceptibility testing (DST) helps reduce the time to AST... more

Background: Conventional methods for isolation and antibiotic susceptibility reports of bacterial isolates from positive blood culture bottles takes at least 48 hours. Direct susceptibility testing (DST) helps reduce the time to AST results by 24 hours enabling early initiation of treatment. Materials and Methods: Blood cultures flagged positive by the Bactec or BacT ALERT systems between July and October 2016 were analyzed by Grams stain. Direct susceptibility test was performed on the broths with monomicrobial growth using the guidelines prescribed by British Society for Antimicrobial Chemotherapy (BSAC). Simultaneously routine culture and susceptibility testing were also carried out for all the isolates. Results: Both gram negative and gram positive organisms were subjected to DST. Out of 30 Gram positive isolates subjected to DST, (n=6) were Enterococcus species and (n=24) Staphylococcus species. Out of 30 Gram negative isolates subjected to DST,the species tested were Klebsiella spp (n=6), Escherichia coli (n=5), Acinetobacter spp (n=7), Pseudomonas (n=1), Morganella (n=1), Salmonella spp (n=10) in number. Conclusions: Direct susceptibility testing enables clinicians to start antimicrobial therapy 24hrs earlier than the routine culture and susceptibility testing, thus reducing the mortality and morbidity in patients with blood stream infections. A high level of concordance was seen between DST and reference method.

Interaction thermodynamics between warfarin, a very popular anticoagulant, and Sudlow I binding site of human (HSA) or bovine (BSA) serum albumin have been examined in strictly controlled experimental conditions (HEPES buffer 50 mM, pH... more

Interaction thermodynamics between warfarin, a very popular anticoagulant, and Sudlow I binding site of human (HSA) or bovine (BSA) serum albumin have been examined in strictly controlled experimental conditions (HEPES buffer 50 mM, pH 7.4 and 25 °C) by means of isothermal titration calorimetry (ITC), fluorescence spectrometry (FS) and frontal analysis capillary electrophoresis (FA/CE). Each technique is based on measurements of a different property of the biochemical system, and then the results allow a critical discussion about the suitability of each approach to estimate the drug-protein binding parameters. The strongest interaction step is properly evaluated by the three assayed approaches being the derived binding constants strongly consistent: from 4 × 104 to 7 × 104 for HSA and from 0.8 × 105 to 1.2 × 105 for BSA. Binding enthalpy variations also show consistent results: -5.4 and -5.6 Kcal/mol for HSA and -4.3 and -3.7 Kcal/mol for BSA, as measured by ITC and FS, respectively...

4-Hydroxynonenal (HNE) is a major aldehydic product formed by peroxidation of omega 6-unsaturated fatty acids and is regarded as a specific marker of lipid peroxidation. In this paper we demonstrate that there is a physiological... more

4-Hydroxynonenal (HNE) is a major aldehydic product formed by peroxidation of omega 6-unsaturated fatty acids and is regarded as a specific marker of lipid peroxidation. In this paper we demonstrate that there is a physiological steady-state concentration of HNE in human venous blood plasma. For the quantitative determination of HNE a modified version of an existing, but tedious and time-consuming HPLC method was developed. The extraction of aldehydic hydrazones from plasma was performed using an Extrelut column and the separation step by thin-layer chromatography was replaced by column chromatography on silica gel. The concentration of HNE in human blood plasma was in the same range as the concentration that was found to inhibit the proliferation of cells of the peripheral tissues, i.e., endothelial cells and fibroblasts in vitro. In an experiment with reduced peripheral blood flow a temporary significant increase of HNE was observed during reperfusion. It was concluded that lipid ...

Lab Tests Online is a "peer-reviewed, non-commercial, patient-centered" resource where patients and their relatives and caregivers can learn about the tests used to screen for, diagnose, and manage disease. Consumers are becoming... more

Lab Tests Online is a "peer-reviewed, non-commercial, patient-centered" resource where patients and their relatives and caregivers can learn about the tests used to screen for, diagnose, and manage disease. Consumers are becoming increasingly involved in the management of their own health care and increasingly have access to their laboratory results through electronic health records. Research has shown that consumers have difficulty with health literacy in general and with numerical data in particular. The Lab Tests Online global websites are an important step toward helping consumers understand the complexity of the pathology process, the expertise of the people involved and the meaning of the results provided to them and their healthcare professionals.

Objective and rationale: Reference intervals provided on laboratory reports are essential for appropriate interpretation of test results, and can significantly impact clinical decision-making and the quality of patient care. Careful... more

Objective and rationale: Reference intervals provided on laboratory reports are essential for appropriate interpretation of test results, and can significantly impact clinical decision-making and the quality of patient care. Careful determination and/or validation of reference intervals by the laboratory for use in the patient population it serves are therefore important to ensure their proper utility. Unfortunately, critical gaps currently exist in accurate and up-to-date pediatric reference intervals for accurate interpretation of laboratory tests performed in children and adolescents. These critical gaps in the available pediatric laboratory reference intervals have the clear potential of contributing to erroneous diagnosis or misdiagnosis of many diseases of childhood and adolescence. Most of the available "normal" ranges for laboratory tests were determined over 2 decades ago on older instruments and technologies, and are no longer relevant considering the current testing technology used by clinical laboratories. It is thus critical and of utmost urgency that a more acceptable and comprehensive database be established. Discussion and conclusion: In the present review, we discuss the considerations and challenges faced when generating and validating reference intervals in accordance to the current guidelines published by the Clinical Laboratory Standards Institute (CLSI). We raise particular attention to the present-day deficiencies in available pediatric reference intervals, and highlight the special issues and unique difficulties that are additionally faced when establishing reference intervals in children. Finally, we highlight a recent Canadian initiative, the CALIPER project, whose mandate is to establish and maintain a database of comprehensive and up-to-date pediatric reference intervals to be eventually made available to all clinical laboratories worldwide.

Here we describe a novel method to conjugate pneumococcal polysaccharides (PnPS) to Luminex microspheres for use in serological assays. 4-(4,6-dimethoxy[1,3,5]triazin-2-yl)-4-methyl-morpholinium (DMTMM) modification of PnPS and... more

Here we describe a novel method to conjugate pneumococcal polysaccharides (PnPS) to Luminex microspheres for use in serological assays. 4-(4,6-dimethoxy[1,3,5]triazin-2-yl)-4-methyl-morpholinium (DMTMM) modification of PnPS and conjugation to carboxyl functional groups on Luminex microspheres (COOH-DMTMM method) was shown to be a reproducible chemistry that efficiently conjugated PnPS to Luminex microspheres without affecting the antigenicity of a broad set of PnPS. The COOH-DMTMM method was compared to three other methods for robustness, reproducibility and effect on PnPS antigenicity in a multiplexed assay format. The other methods examined included adsorption of the unmodified PnPS to Luminex microspheres, oxidation of the PnPS to conjugate them to amino-modified microspheres using carbodiimide chemistry and poly-l-lysine modification of the PnPS before conjugating to carboxy Luminex microspheres using carbodiimide chemistry. Of the four methods, the COOH-DMTMM chemistry was shown to be a robust methodology, producing stable PnPS coupled microspheres with a 4-log dynamic range and low cross-reactivity when used in a PnPS-specific IgG serology assay. This novel chemistry should be useful for developing serological assays to measure antibodies to polysaccharides for use in vaccine and epidemiology studies.

The purpose of measurement standardization is to achieve closer comparability of results obtained using different commercial systems. Regarding serum protein immunoassays, a reference preparation (BCR-470) was released in 1993 and adopted... more

The purpose of measurement standardization is to achieve closer comparability of results obtained using different commercial systems. Regarding serum protein immunoassays, a reference preparation (BCR-470) was released in 1993 and adopted by manufacturers across the world to value-assign their assay calibrators for routine methods to reduce method-dependent variation. Moving from nephelometric (Beckman Immage 800) to turbidimetric determination (Roche Cobas c 501) of seven serum proteins, we preliminarily checked the comparability of results between the two systems. The study was performed according to the CLSI EP9-A protocol on 30 fresh sera, tested on each system in duplicate, and subdivided on two different days, without recalibration and using manufacturers' control materials to validate the runs. Both manufacturers' package inserts provide statements that kit calibrators are traceable to BCR-470. Suggested reference intervals are also the same. Although a fairly good correlation was observed (r=0.955), the comparison of ceruloplasmin methods produced evidence of highly significant proportion-al (regression slope, 0.572) and constant bias (intercept, 0.05 g/L). Absolute and percentage mean differences were −0.11 g/L (95% confidence interval (CI) −0.13 to −0.10 g/L) and −39.1% (CI −43.1 to −35.2%), respectively. No other evaluated proteins showed similar problems. Lacking a ceruloplasmin reference method, it is impossible to demonstrate that one of the two assays produces true ceruloplasmin values. The problem is, however, that results coming from the two assays are clearly not comparable. This may be either due to a lack of commutability of the reference material with biological samples in the evaluated assays or to calibration problems by manufacturers in one of the stages of the calibration hierarchy.

The content of phenolic compounds determines the state of phenolic ripening of red grapes, which is a key criterion in setting the harvest date to produce quality red wines. Wine phenolics are also important quality components that... more

The content of phenolic compounds determines the state of phenolic ripening of red grapes, which is a key criterion in setting the harvest date to produce quality red wines. Wine phenolics are also important quality components that contribute to the color, taste, and mouth feel of wines. Spectroscopic techniques (e.g., near and mid infrared) offer the potential to simplify and reduce the analytical time for a range of grape and wine analytes. It is this characteristic, together with the ability to simultaneously measure several analytes in the same sample at the same time, which makes these techniques very attractive for use in both industry and research. The objective of this mini review is to present examples and to discuss different applications of visible (VIS), near infrared (NIR) and mid infrared (MIR) to assess and measure phenolic compounds in grape and wines.

Screening and diagnostic criteria for gestational diabetes (GDM) are inconsistent across Europe, and the development of a uniform GDM screening strategy is necessary. Such a strategy would create opportunities for more women to receive... more

Screening and diagnostic criteria for gestational diabetes (GDM) are inconsistent across Europe, and the development of a uniform GDM screening strategy is necessary. Such a strategy would create opportunities for more women to receive timely treatment for GDM. Developing a consensus on screening for GDM in Europe is challenging, as populations are diverse and healthcare delivery systems also differ. The European Board & College of Obstetrics and Gynaecology (EBCOG) has responded to this challenge by appointing a steering committee, including members of the EBCOG and the Diabetic Pregnancy Study Group (DPSG) associated with the EASD, to develop a proposal for the use of uniform diagnostic criteria for GDM in Europe. A proposal has been developed and has now been approved by the Council of the EBCOG. The current proposal is to screen for overt diabetes at the first prenatal contact using cutoff values for diabetes outside pregnancy, with particular efforts made to screen high-risk groups. When screening for GDM is performed at 24 weeks' gestation or later, the proposal is now to use the 75 g OGTT with the new WHO diagnostic criteria for GDM. However, more research is necessary to evaluate the best GDM screening strategy for different populations in Europe. Therefore, no clear recommendation has been made on whether a universal one-step, two-step or a risk-factorbased screening approach should be used. The use of the same WHO diagnostic GDM criteria across Europe will be an important step towards uniformity.

The Society of Radiologists in Ultrasound convened a panel of specialists from radiology, hepatology, pathology, and basic science and physics to arrive at a consensus regarding the use of elastography in the assessment of liver fibrosis... more

The Society of Radiologists in Ultrasound convened a panel of specialists from radiology, hepatology, pathology, and basic science and physics to arrive at a consensus regarding the use of elastography in the assessment of liver fibrosis in chronic liver disease. The panel met in Denver, Colo, on October 21-22, 2014, and drafted this consensus statement. The recommendations in this statement are based on analysis of current literature and common practice strategies and are thought to represent a reasonable approach to the noninvasive assessment of diffuse liver fibrosis. (©) RSNA, 2015 Online supplemental material is available for this article.

CONTEXT: Thyroid uptake and scintigraphy using 99mTc-pertechnetate has proven to be more advantageous than with 131I-iodide, since the images have better quality, the procedure is faster and the patient is submitted to a lower radiation... more

CONTEXT: Thyroid uptake and scintigraphy using 99mTc-pertechnetate has proven to be more advantageous than with 131I-iodide, since the images have better quality, the procedure is faster and the patient is submitted to a lower radiation dose. OBJECTIVE: The purpose of this study was to standardize a simple and fast methodology for performing thyroid uptake and scintigraphy and to determine the normal values for 99mTc- pertechnetate uptake. TYPE OF STUDY: Prospective, non-randomized. SETTING: Division of Nuclear Medicine, Department of Radiology, School of Medical Sciences, Campinas State University. PARTICIPANTS: The study consisted of 47 normal individuals, 30 women and 17 men, with ages ranging from 19 to 61 years (mean of 33 years). PROCEDURES: The laboratory assessment of thyroid function consisted of serum dosages of ultra-sensitive thyroxin and thyrotrophin. Twenty minutes after an intravenous injection of 10 mCi (370 MBq) of 99mTc-pertechnetate, the images were obtained on a ...

Timely communication of significant or unexpected findings in surgical pathology can significantly improve patient care. Although surgical pathology critical values have been published, no systematic assessment in pediatric surgical... more

Timely communication of significant or unexpected findings in surgical pathology can significantly improve patient care. Although surgical pathology critical values have been published, no systematic assessment in pediatric surgical pathology has been published. We surveyed pediatric pathologists and pediatric subspecialists to develop pediatric surgical pathology critical values for verbal reporting before the final pathology report. A policy and process for reporting and documentation was implemented, with retrospective and prospective quality review. Critical values cases constituted 9.4% of surgical pathology accessions. Retrospective analysis revealed that 80% (73/91) had been reported and documented before policy implementation. Following implementation, 97.3% (402/413) were verbally reported and documented. A multidisciplinary group provided valuable information about critical values that might not have been obvious to pediatric pathologists but are important for patient care. Although the term critical values has become embedded in the surgical pathology literature, we would propose an alternative term for significant or unexpected findings that require timely communication and documentation.

Quantitation of exact total protein content is often a key step and is common to many applications in general biochemistry research and routine clinical laboratory practice. Before embarking on any type of protein analysis, particularly... more

Quantitation of exact total protein content is often a key step and is common to many applications in general biochemistry research and routine clinical laboratory practice. Before embarking on any type of protein analysis, particularly comparative techniques, it is important to accurately quantitate the amount of protein in the sample. In order to assess the quality of total protein estimation results, five methods were tested and were applied to the same pooled plasma sample. For this aim, Bradford (Coomassie Brilliant Blue), Lowry (Folin-Ciocalteau), Biüret, Pesce and Strande (Ponceau-S/TCA), and modified method of Schaffner-Weismann (Amido Black 10B) were used. The last two methods employ simultaneous precipitation of proteins with the acid containing dye solutions followed by dissolution of precipitate in a NaOH solution. It is shown that each assay has advantages and disadvantages relative to sensitivity, ease of performance, acceptance in literature, accuracy and reproducibility/coefficient of variation. All of the methods tested show a CV % b 6. Besides pooled plasma, a known concentration of human serum albumin was also analyzed and discussed by means of standardization of plasma total protein content.

Background: The development of multiparameter flow cytometry (FCM) and increasingly sophisticated analysis software has considerably improved the exploration of hematological disorders. These tools have been widely applied in leukaemias,... more

Background: The development of multiparameter flow cytometry (FCM) and increasingly sophisticated analysis software has considerably improved the exploration of hematological disorders. These tools have been widely applied in leukaemias, lymphomas, and myelodysplasias, yet with very heterogeneous approaches. Consequently, there is no extensive reference document reporting on the characteristics of normal human bone marrow (BM) in multiparameter FCM. Here, we report a reference analysis procedure using relevant antibody combinations in normal human BM. Methods: A first panel of 23 antibodies, constructed after literature review, was tested in four-color combinations (including CD45 in each) on 30 samples of BM. After evaluation of the data, a second set of 22 antibodies was further applied to another 35 BM samples. All list-modes from the 65 bone marrow samples were reviewed collectively. A systematised protocol for data analysis was established including biparametric representations and color codes for the three major lineages and undifferentiated cells. Results: This strategy has allowed to obtain a reference atlas of relevant patterns of differentiation antigens expression in normal human BM that is available within the European LeukemiaNet. This manuscript describes how this atlas was constructed. Conclusions: Both the strategy and atlas could prove very useful as a reference of normality, for the determination of leukemia-associated immunophenotypic patterns, analysis of myelodysplasia and, ultimately, investigation of minimal residual disease in the BM. V

Clotrimazole is a wide spectrum local imidazolic antifungal agent used in several dermatological creams, having e.g. 1% (m/m) such as Canesten 1 and Fungizid-ratiopharm 1 cream. In the present work, a new system based on solid lipid... more

Clotrimazole is a wide spectrum local imidazolic antifungal agent used in several dermatological creams, having e.g. 1% (m/m) such as Canesten 1 and Fungizid-ratiopharm 1 cream. In the present work, a new system based on solid lipid nanoparticles (SLN TM) containing the identical concentration of drug has been developed. A comparative study between the rheological properties of the referred creams and the developed aqueous SLN dispersions was carried out. The influence of incorporation of SLN in a standard hydrophilic cream on its flow curves was also assessed. In addition, the release of clotrimazole from the two commercial creams, as well as from aqueous SLN dispersions was studied. Concerning the rheological investigations, all tested commercial creams revealed very low shear rates and no yield points. Lipid nanoparticles having a mean diameter of approx. 200 nm have been incorporated into a hydrophilic cream, in a concentration of 20%, 30% or 40% (m/m). The hydrophilic cream containing 20% of SLN showed a dilatant-like character; however, increasing the percentage of incorporated lipid nanoparticles to 30% and 40% the formulation changed to a more pseudoplastic character, showing yield values of 28 Pa and 39 Pa, respectively. For in vitro release studies, Franz diffusion cells with a cellulose acetate membrane were used to measure the release of clotrimazole from two different commercial formulations in comparison to the aqueous SLN dispersion. After 6 h the amount of drug released was higher than 48% when delivered from both investigated commercial formulations and not higher than 25% when delivered from the aqueous SLN dispersion. The percentage of drug released determined after 24 h was more than 50% for Canesten 1 cream and Fungizidratiopharm 1 cream and not higher than 30% for the developed SLN formulation showing its prolonged release character.

90 Y microspheres are important therapeutic radiopharmaceuticals used in the treatment of liver cancer through a process known as selective internal radiation therapy. SIR-spheres s is a radiopharmaceutical product that is comprised of 90... more

90 Y microspheres are important therapeutic radiopharmaceuticals used in the treatment of liver cancer through a process known as selective internal radiation therapy. SIR-spheres s is a radiopharmaceutical product that is comprised of 90 Y microspheres suspended in sterile, pyrogen-free water for injection into patients. It is necessary to establish for the SIR-spheres s production the capability of accurately measuring the activity of this product to a traceable national measurement standard. An activity standard for SIR-spheres s was developed from a standard for 90 Y solution, employing a highly quantifiable chemical digestion process. Calibration factors for the manufacturer's ionisation chambers were determined for 1 and 5 ml of the SIR-spheres s product placed in Wheaton vials, for both 34% and 44% of 90 Y microsphere concentration. r

Introducción. La implantación de una prótesis total de cadera (PTC) es un procedimiento habitual en cirugía ortopédica, que frecuentemente precisa reposición hemática en el postoperatorio inmediato. El porcentaje de pacientes protetizados... more

Introducción. La implantación de una prótesis total de cadera (PTC) es un procedimiento habitual en cirugía ortopédica, que frecuentemente precisa reposición hemática en el postoperatorio inmediato. El porcentaje de pacientes protetizados que recibe una transfusión homóloga oscila en torno al 60% (1). Sin embargo, esta práctica no está exenta de riesgos; se asocia a un riesgo de transmisión de enfermedades infeccionas, riesgo de aloinmunización, reacciones hemolíticas o febriles por aloanticuerpos, reacciones injerto contra huésped y se le atribuye mayor incidencia de infecciones y una estancia hospitalaria más prolongada (1,2). Además, es un recurso caro y no siempre disponible. Con objeto de minimizar las necesidades de transfusión homóloga en pacientes que iban a ser intervenidos de prótesis total de cadera se diseñó un programa de ahorro de sangre basado en la autodonación prequirúrgica en pacientes que cumplieran los criterios de selección. Entre 1996 y 2001 se incluyeron en este programa 220 pacientes, consiguiéndose

Hand 13 C-NMR (CD 3 OD, Table 1) spectra of 2 showed signals assignable to two methylenes and a methine bearing an oxygen function [d 3.70 (2H, m, 3-H 2), 4.01 (1H, m, 2-H), 4.26, 4.31 (1H each, both dd, Jϭ5.2, 11.9 Hz, 1-H 2)] and an... more

Hand 13 C-NMR (CD 3 OD, Table 1) spectra of 2 showed signals assignable to two methylenes and a methine bearing an oxygen function [d 3.70 (2H, m, 3-H 2), 4.01 (1H, m, 2-H), 4.26, 4.31 (1H each, both dd, Jϭ5.2, 11.9 Hz, 1-H 2)] and an angeloyl moiety [d 1.90 (3H, br s, 5Ј-H 3), 1.98 (3H, dd, Jϭ1.5, 7.3 Hz, 4Ј-H 3), 6.14 (1H, m, 3Ј-H)] together with a b-D-glucopyranosyl and a b-D-apiofuranosyl parts [d 4.43 (1H, d, Jϭ8.0 Hz, 1Љ-H), 5.00 (1H, d, Jϭ2.5 Hz, 1ٞ-H)]. Finally, the connectivities of the acyl group and the glycosyl linkages in 2 were elucidated on the basis of HMBC experiment, which showed long-range correlations between the following proton and carbon pairs as shown in Fig. 1: the 1-protons and the angeloyl ester carbonyl carbon (d C 169.1, 1Ј-C), the Glc-1-proton (1Љ-H) and the 2-carbon (d C 79.9), and the Api-1-proton (1ٞ-H) and the Glc-6-carbon (d C 68.9, 6Љ-C). Consequently, everlastoside G was determined to be 1-Oangeloylglycerol 2-O-b-D-apiofuranosyl-(1→6)-b-D-glucopy

A simple and sensitive gas chromatographic method was designed for quantitative analysis of Streptococcus pneumoniae capsular polysaccharides, activated polysaccharides, and polysaccharide conjugates. Pneumococcal serotypes 1, 3, 4, 5,... more

A simple and sensitive gas chromatographic method was designed for quantitative analysis of Streptococcus pneumoniae capsular polysaccharides, activated polysaccharides, and polysaccharide conjugates. Pneumococcal serotypes 1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, and 23F polysaccharide or conjugate were subjected to methanolysis in 3 N hydrochloric acid in methanol followed by re-Nacetylation and trimethylsilylation. Derivatized samples were chromatographed and detected using gas chromatography with mass selective detector. Gas chromatographic results were compared with colorimetric values with agreement of 92 to 123% over the range of all samples tested. Monosaccharides released during methanolysis included hexoses, uronic acids, 6-deoxy-hexoses, amino sugars, and alditols. Quantitative recovery of monosaccharides was achieved for all serotypes by the use of a single methanolysis, derivatization, and chromatography procedure. Response factors generated from authentic monosaccharide standards were used for quantitation of pneumococcal polysaccharides and conjugates with conWrmation of peak assignments by retention time and mass spectral analysis. This method allows saccharide quantitation in multivalent pneumococcal vaccine intermediates and Wnal drug products with low-level detection (10 pg) and peak purity.

A major factor in the direct determination of lutein in spinach extracts proved to be obtaining reproducible and stable chromatography of lutein. This was achieved on a C30 column with the mobile phase acetone-0.1 M triethylammonium... more

A major factor in the direct determination of lutein in spinach extracts proved to be obtaining reproducible and stable chromatography of lutein. This was achieved on a C30 column with the mobile phase acetone-0.1 M triethylammonium acetate (TEAA) buffer (pH 7) 9:1 (v/v). Extraction of 10 mg of lyophilized spinach with 10 mL of extraction solvent (ethanol, acetone, ethanol-ethyl acetate 1:1 (v/v), methanol-THF 1:1 (v/v)) for 15 min with magnetic stirring under nitrogen resulted in equal yields of lutein. The yields were enhanced by addition of 15% of 1 M TEAA buffer pH 7 to all four extraction solvents. As confirmed by recovery experiments, no loss of lutein occurred during the extraction. The relative standard deviation from triplicate extractions was less than 5%. The addition of 15% TEAA pH 7 to acetone enhanced the extraction yield of lutein also from unlyophilized spinach. The content of lutein in different spinach samples ranged from 5 to 15 mg/100 g of fresh weight. The first separation is reported of all the carotenoids and chlorophylls on a C18 core-shell column and the addition of 15% of 1 M TEAA buffer pH 7 to acetone also enhanced the extraction yield of ␤-carotene compared to the yield produced by pure acetone.

Accurate estimates of energy and nutrient intake of individuals and populations are contingent on the reliability of information obtained about food intake, food composition, and portion size. This article reviews issues related to the... more

Accurate estimates of energy and nutrient intake of individuals and populations are contingent on the reliability of information obtained about food intake, food composition, and portion size. This article reviews issues related to the definition, determination, and use of portion sizes in dietary assessment and public education. Food frequency questionnaires USDA FDA Pyramidz2 Food labelzx Willett et al.b.3z Block et al.2.33 Grains and cereals Bread Crackers Rice, cooked Cereal, ready-to-eat Cereal, cooked Whole fruit Juice Fresh, frozen, or canned Raw leafy Carrots, raw Potato French fries Tomato juice Beef Hamburger Poultry Fish Dairy Milk Cheese Fats, sweets Butter Cookies Doughnuts Alcohol Wine Fruit Vegetables Meat 1 slice 3-4 small 4 oz 1 medium elass I a Standard portion sizes vary by weight and size of items. Harvard University Health Professionals Follow-up Study, 1990 questionnaire. Food Guides Historical Background. For nearly a century, the USDA has published food guides to help the public select healthful diets.I5 In its earliest efforts to suggest portion sizes, the agency recommended that families purchase specific quantities of foods to supply nutrients then known to be essential. Quantities were expressed by weight, volume, or 100kcal portions; 80,000 kcal (800 100-kcal portions) was the figure used to meet the weekly needs of an average family of two adults and three children. USDA advised families to obtain these portions through specific food purchases, such as 14 lb potatoes, 2 lb turnips, and 14 qt milk per week.16 Later, USDA specified yearly amounts of foods needed by men and women.17 Although portion sizes could be estimated from such figures, they were not stated explicitly. USDA first defined daily portions during World War 11. It recommended consumption of one or two

Background aims. The clinical applications of in vitro manipulated cultured cells and their precursors are often made use of in therapeutic trials. However, tissue cultures can be easily contaminated by the ubiquitous Mollicutes... more

Background aims. The clinical applications of in vitro manipulated cultured cells and their precursors are often made use of in therapeutic trials. However, tissue cultures can be easily contaminated by the ubiquitous Mollicutes micro-organisms, which can cause various and severe alterations in cellular function. Thus methods able to detect and trace Mollicutes impurities contaminating cell cultures are required before starting any attempt to grow cells under good manufacturing practice (GMP) conditions. Methods. We developed a multiplex quantitative polymerase chain reaction (qPCR) assay specifi c for the 16S -23S rRNA intergenic spacer regions, for the Tuf and P1 cytoadhesin genes, able to detect contaminant Mollicutes species in a single tube reaction. The system was validated by analyzing different cell lines and the positive samples were confi rmed by 16S and P1 cytoadhesin gene dideoxy sequencing. Results. Our multiplex qPCR detection system was able to reach a sensitivity, specifi city and robustness comparable with the culture and the indicator cell culture method, as required by the European Pharmacopoeia guidelines. Conclusions. We have developed a multiplex qPCR method, validated following International Conference on Harmonization (ICH) guidelines, as a qualitative limit test for impurities, assessing the validation characteristics of limit of detection and specifi city. It also follows the European Pharmacopoeia guidelines and Food and Drug Administration (FDA) requirements.

Following the approach of the National Primary Laboratory of the UK (NPL) for the calibration of radionuclide calibrators, but using a commercially available instrument with no data available in the literature, the radionuclide calibrator... more

Following the approach of the National Primary Laboratory of the UK (NPL) for the calibration of radionuclide calibrators, but using a commercially available instrument with no data available in the literature, the radionuclide calibrator response was investigated as a function of different measurement geometries at the ''Regina Elena'' National Cancer Institute (IRE) in Rome.

A collaborative study involving a large sample of European Americans was typed for the histocompatibility loci of the HLA DR-DQ region and subjected to intensive typing validation measures in order to accurately determine haplotype... more

A collaborative study involving a large sample of European Americans was typed for the histocompatibility loci of the HLA DR-DQ region and subjected to intensive typing validation measures in order to accurately determine haplotype composition and frequency. The resulting tables have immediate application to HLA typing and allogeneic transplantation. The loci within the DR-DQ region are especially valuable for such an undertaking because of their tight linkage and high linkage disequilibrium. The 3798 haplotypes, derived from 1899 unrelated individuals, had a total of 75 distinct DRB1-DQA1-DQB1 haplotypes. The frequency distribution of the haplotypes was right skewed with haplotypes occurring at a frequency of less than 1% numbering 59 and yet constituting less than 12% of the total sample. Given DRB1 typing, it was possible to infer the exact DQA1 and DQB1 composition of a haplotype with high confidence (>90% likelihood) in 21 of the 35 highresolution DRB1 alleles present in the sample. Of the DRB1 alleles without high reliability for DQ haplotype inference, only *0401, *0701 and *1302 were common, the remaining 11 DRB1 alleles constituting less than 5% of the total sample. This approach failed for the 13 serologically equivalent DR alleles in which only 33% of DQ haplotypes could be reliably inferred. The 36 DQA1-DQB1 haplotypes present in the total sample conformed to the known pattern of permissible heterodimers. Four DQA1-DQB1 haplotypes, all rare, are reported here for the first time. The haplotype frequency tables are suitable as a reference standard for HLA typing of the DR and DQ loci in European Americans.

We present two different methods for determining levels of glutathione in complex biological samples and plasma. The DTNB/GR enzyme recycling method is sensitive and requires no specialized equipment. The HPLC method is particularly... more

We present two different methods for determining levels of glutathione in complex biological samples and plasma. The DTNB/GR enzyme recycling method is sensitive and requires no specialized equipment. The HPLC method is particularly useful for situations in which sample amounts are limited. Detailed instructions for performing each method as well as the advantages and disadvantages of each are discussed in this chapter.

Background: Hepatitis delta virus (HDV) RNA viral load measurement is critical in diagnosis and monitoring the response to antiviral treatment. Objectives: Our aim is to design a real time PCR method for accurate quantitation of HDV RNA... more

Background: Hepatitis delta virus (HDV) RNA viral load measurement is critical in diagnosis and monitoring the response to antiviral treatment. Objectives: Our aim is to design a real time PCR method for accurate quantitation of HDV RNA in clinical specimens using an armored RNA as external standard, and an intrinsic internal control. Study design: A plasmid bearing delta antigen region of genotype I HDV genome was used to develop an armored RNA. Serial dilutions of the armored HDV RNA standard with 10 12 copy/mL were used as standards for quantitation. A primer-probe set derived from HDAg region was used in one step EZ RT PCR kit chemistry which uses rTth enzyme allowing reverse transcription and polymerization in the same tube. The kit also uses the advantage of uracil-N-glycosylase (UNG) enzyme treatment to prevent PCR contamination. Results: The established assay has a dynamic range of 10 2 -10 11 copy/mL with a PCR efficiency of 96.9%. Detection limit was 858 ± 32 copy/mL with 95% confidence interval. Intra-and inter-assay variabilities were low for high, medium and low levels of viremia. Incorporation of freely circulating GAPDH in serum into the assay as an intrinsic internal control prevented false negative results and failures in PCR amplifications due to inhibitors, inefficient extraction procedures or enzymatic reactions. Conclusion: In conclusion, this study defines a novel assay for sensitive and reliable quantification of HDV RNA using an armored HDV RNA as a standard and GAPDH in plasma or serum as an intrinsic internal control in a single tube.

Different technologies, such as quantitative real-time PCR or microarrays, have been developed to measure microRNA (miRNA) expression levels. Quantification of miRNA transcripts implicates data normalization using endogenous and exogenous... more

Different technologies, such as quantitative real-time PCR or microarrays, have been developed to measure microRNA (miRNA) expression levels. Quantification of miRNA transcripts implicates data normalization using endogenous and exogenous reference genes for data correction. However, there is no consensus about an optimal normalization strategy. The choice of a reference gene remains problematic and can have a serious impact on the actual available transcript levels and, consequently, on the biological interpretation of data. In this review article we discuss the reliability of the use of small RNAs, commonly reported in the literature as miRNA expression normalizers, and compare different strategies used for data normalization. A workflow strategy is proposed for normalization of miRNA expression data in an attempt to provide a basis for the establishment of a global standard procedure that will allow comparison across studies.

Different age structures in two populations complicate any comparison of their levels of mortality. Many methods exist which provide death rates or mortality indices adjusted for age and other factors. Such summary measures inevitably... more

Different age structures in two populations complicate any comparison of their levels of mortality. Many methods exist which provide death rates or mortality indices adjusted for age and other factors. Such summary measures inevitably lose information, but they are useful for the initial examination of large quantities of data and for the presentation of results. This paper reviews a number of techniques available for producing age-adjusted death rates or mortality indices, emphasizing their historical development. Formulae are given for their calculation. The appropriate context for using each method, and its associated disadvantages are described.

Background: A quality assessment tool for diagnostic accuracy studies, named QUADAS, has recently been developed. Although QUADAS has been used in several systematic reviews, it has not been formally validated. The objective was to... more

Background: A quality assessment tool for diagnostic accuracy studies, named QUADAS, has recently been developed. Although QUADAS has been used in several systematic reviews, it has not been formally validated. The objective was to evaluate the validity and usefulness of QUADAS. Methods: Three reviewers independently rated the quality of 30 studies using QUADAS. We assessed the proportion of agreements between each reviewer and the final consensus rating. This was done for all QUADAS items combined and for each individual item. Twenty reviewers who had used QUADAS in their reviews completed a short structured questionnaire on their experience of QUADAS. Results: Over all items, the agreements between each reviewer and the final consensus rating were 91%, 90% and 85%. The results for individual QUADAS items varied between 50% and 100% with a median value of 90%. Items related to uninterpretable test results and withdrawals led to the most disagreements. The feedback on the content of the tool was generally positive with only small numbers of reviewers reporting problems with coverage, ease of use, clarity of instructions and validity. Conclusion: Major modifications to the content of QUADAS itself are not necessary. The evaluation highlighted particular difficulties in scoring the items on uninterpretable results and withdrawals. Revised guidelines for scoring these items are proposed. It is essential that reviewers tailor guidelines for scoring items to their review, and ensure that all reviewers are clear on how to score studies. Reviewers should consider whether all QUADAS items are relevant to their review, and whether additional quality items should be assessed as part of their review.

In diabetes research, mouse and rat models are used for in vivo experiments, and quantification of insulin in serum samples under different pathophysiological conditions and after treatment with compounds is essential. There are few... more

In diabetes research, mouse and rat models are used for in vivo experiments, and quantification of insulin in serum samples under different pathophysiological conditions and after treatment with compounds is essential. There are few commercial radioimmunoassay and enzyme-linked immunosorbent assay (ELISA) assay kits to determine the rat/mouse plasma levels of insulin. However, reliability in insulin measurements using the available biological assays is a great concern. The authors report a robust, extremely sensitive electrochemiluminescence (ECL) insulin assay using the Origen technology platform. The assay performance, as judged by the Z′ value of 0.82±0.03 and the signal-to-background (S/B) ratio of 133, suggests that this is a robust and reliable assay. The intra-assay and interassay variation is less than 5%. The dynamic range of detection for insulin is 5 pg to 5 ng in the ECL assays. Recovery of insulin was about 100% when different volumes of serum were spiked with exogenous...

Streptococcus agalactiae, group B streptococci (GBS) is the leading cause of severe bacterial infections in newborns. GBS expression studies allowed the identification and characterization of virulence factors and a better understanding... more

Streptococcus agalactiae, group B streptococci (GBS) is the leading cause of severe bacterial infections in newborns. GBS expression studies allowed the identification and characterization of virulence factors and a better understanding of the host-pathogen-environment interactions. The measurement of transcript levels by quantitative real-time PCR (qRT-PCR) is a widely used technique in GBS; however, a systematic evaluation and validation of reference gene stability for normalization purposes in GBS expression studies is currently lacking. Therefore, we analyzed the stability of 10 candidate reference genes (16SrRNA, glcK, glnA, groEL, gyrA, recA, rpoB, rpsL, sdhA and tkt) in three GBS prototype strains (O90R, NEM316 and 2603V/R) grown at different temperature conditions (37°C and 40°C). Our approach was based on the calibration of transcript levels from each gene against the number of bacteria from the same sample (ratio messenger RNA/genomic DNA). As a complementary analysis, reference gene stability was also investigated through the bioinformatic applications, geNorm and NormFinder. Considering the whole GBS development cycle, only a minority of genes were stable under both growth conditions, but this number increased when restricting the analysis to the logarithmic time-points. The range of stable genes was higher at 37°C, where recA and sdhA were stable simultaneously for the three strains, and six out of 10 genes were stable for at least two strains. At 40°C, recA showed up again as one of the best options, suggesting its potential use as reference gene in future qRT-PCR studies. The results generated with geNorm and NormFinder were consistent with those obtained experimentally and evidenced minor variations either among strains or temperature conditions. In conclusion, the fluctuation of expression of reference genes observed among different GBS strains and growth conditions highlights the importance of carefully validating, for each experimental scenario, the use of reference genes for qRT-PCR normalization purposes. Nevertheless, recA seems to be a good candidate for such optimizations.

Several inborn errors of metabolism with abnormal polyol concentrations in body fluids are known to date. Most of these defects can be diagnosed by the assessment of urinary concentrations of polyols. We present two methods using tandem... more

Several inborn errors of metabolism with abnormal polyol concentrations in body fluids are known to date. Most of these defects can be diagnosed by the assessment of urinary concentrations of polyols. We present two methods using tandem mass spectrometry for screening for inborn errors affecting polyol metabolism. Urine samples supplemented with internal standards ([ 13 C 4 ]erythritol, [ 13 C 2 ]arabitol and [ 2 H 3 ]sorbitol) were desalted by a mixed-bed ion-exchange resin. Separation was achieved by two different columns. Sugar isomers could not be separated using a Prevail Carbohydrate ES 54 column (method 1), whereas with the other column (Aminex HPX-87C) separation of the isomers was achieved (method 2). Multiple reaction monitoring polyol detection was achieved by tandem mass spectrometry with an electron ion-spray source operating in the negative mode. Age-related reference ranges of polyols (erythritol, treitol, arabitol, ribitol, xylitol, galactitol, mannitol, sorbitol, sedoheptitol and perseitol) in urine were established. The applicability of the method was demonstrated by the abnormal polyol concentrations observed in patients with transaldolase deficiency, ribose-5-phosphate isomerase deficiency and classical galactosaemia. This paper describes two methods for the analysis of urinary polyols by liquid chromatography-tandem mass spectrometry. Method 1 is a fast screening method with the quantification of total isomers and method 2 is a more selective method with the separate quantification of the polyols. Both methods can be used for diagnosing inborn errors of metabolism affecting polyol metabolism. Polyols, or polyhydric alcohols, can be formed by the reduction of sugars and are classified on the basis of the numbers of carbon atoms: erythritol and threitol are C 4-polyols (tetritols);

Consensus practices and regulatory guidance for liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) assays of small molecules are more aligned globally than for any of the other bioanalytical techniques addressed by the... more

Consensus practices and regulatory guidance for liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) assays of small molecules are more aligned globally than for any of the other bioanalytical techniques addressed by the Global Bioanalysis Consortium. The three Global Bioanalysis Consortium Harmonization Teams provide recommendations and best practices for areas not yet addressed fully by guidances and consensus for small molecule bioanalysis. Recommendations from all three teams are combined in this report for chromatographic run quality, validation, and sample analysis run acceptance.

The role of computed tomography in PET/CT imaging includes acquisition techniques for diagnostic, anatomical localization and attenuation correction purposes. Diagnostic reference levels of the volumetric computed tomography dose index... more

The role of computed tomography in PET/CT imaging includes acquisition techniques for diagnostic, anatomical localization and attenuation correction purposes. Diagnostic reference levels of the volumetric computed tomography dose index (CTDIvol) are available for dedicated CT procedures of selected body regions but similar reference levels for whole-body CT employed in PET/CT exams are limited. This work reports CTDIvol values from sites that conduct whole-body oncology PET/CT exams and participated in the SNMMI CTN scanner validation program. From 2010 to 2014, a total of 154 sites submitted CT acquisition parameters used in their clinical [(18)F]FDG PET/CT oncology protocols. From these parameters, the CTDIvol was estimated using the ImPACT CTDI dosimetry tables. Histograms of CTDIvol values were created for each year and descriptive statistics, including mean, median and 75th percentile were reported. A repeated measures ANOVA was performed to determine whether significant differ...

In this study, UV-visible spectrophotometry (UV-Vis) and high-performance liquid chromatography (HPLC) were used for simultaneous analysis of chelating agents diethylenetriamine pentaacetic acid (DTPA), ethylenediamine tetraacetic acid... more

In this study, UV-visible spectrophotometry (UV-Vis) and high-performance liquid chromatography (HPLC) were used for simultaneous analysis of chelating agents diethylenetriamine pentaacetic acid (DTPA), ethylenediamine tetraacetic acid (EDTA), and nitrilotriacetic acid (NTA), as their metal chelates in dishwashing detergents, natural waters, and pulp mill water. The total amounts of the chelating agents in dishwashing detergents were verified by potentiometric titration with Fe(III) solution. Nickel(II) chelates were determined by UV-Vis and iron(III)chelates by HPLC and titration. Recoveries of DTPA, EDTA, and NTA from a standard mixture of analytes by UV-Vis were 107±7, 101±12 and 94±13%, respectively, and the recovery of the total amount of complexing agents was 99±4%. The limits of detection for DTPA, EDTA, and NTA were 667, 324, and 739 lmol L À1 , respectively. In HPLC measurements the optimized mobile phase contained 0.03 mol L À1 sodium acetate, 0.002 mol L À1 tetrabutylammonium bromide, and 5% methanol at pH 3.15 and the detection was by UV-Vis detection at 254 nm. All three complexing agents could be separated from each other in a simultaneous analysis in less than 5 min. The limits of detection were 0.34, 0.27, and 0.62 lmol L À1 for DTPA, EDTA, and NTA, respectively. The total amounts of the analytes measured in the dishwashing detergents by the three techniques were found to be highly comparable (ANOVA: F=0.04, P=0.96). R 2 values were 0.99 for EDTA, 0.99 for NTA, and 0.99 for all the results when UV-Vis and HPLC determinations were compared using regression lines. The UV-Vis and HPLC methods were proved to be viable also for analyses of natural and pulp mill waters. The absence of matrix interferences was verified by the standard addition technique.

The Sentence Repetition test (SR) has been used to assess aphasic and other patients in neuropsychological settings. SR has been shown to be reliable and valid, as well as sensitive to patients with brain injury. This study was conducted... more

The Sentence Repetition test (SR) has been used to assess aphasic and other patients in neuropsychological settings. SR has been shown to be reliable and valid, as well as sensitive to patients with brain injury. This study was conducted to update the normative data for the SR. Sensitivity and specificity tests of the SR using the new normative data was also conducted. The findings suggest that SR is sensitive to specific brain dysfunction that affects the ability to repeat sentences as well as attention and concentration. The new normative data show that the ability to repeat sentences is not influenced by age, but education appears to be a significant factor in SR performance. In a clinical group, a reduction of SR performance was related to length of unconsciousness and was more impaired for patients with left-hemisphere injury. Good specificity and sensitivity were found. The findings of this study confirm that with new norms, SR continues to be a useful tool for neuropsychological assessment.