Identical Sample Sets Research Articles

Performance Characteristics of Mass Spectrometry-Based Analytical Procedures for Quantitation of Nitrosamines in Pharmaceuticals: Insights from an Inter-laboratory Study

With the discovery of carcinogenic nitrosamine impurities in pharmaceuticals in 2018 and subsequent regulatory requirements for risk assessment for nitrosamine formation during pharmaceutical manufacturing processes, storage or from contaminated supply chains, effective testing of nitrosamines has become essential to ensure the quality of drug substances and products. Mass spectrometry has been widely applied to detect and quantify trace amounts of nitrosamines in pharmaceuticals. As part of an effort by regulatory authorities to assess the measurement variation in the determination of nitrosamines, an inter-laboratory study was performed by the laboratories from six regulatory agencies with each of the participants using their own analytical procedures to determine the amounts of nitrosamines in a set of identical samples. The results demonstrated that accurate and precise quantitation of trace level nitrosamines can be achieved across multiple analytical procedures and provided insight into the performance characteristics of mass spectrometry-based analytical procedures in terms of accuracy, repeatability and reproducibility.

Measuring Serum Vedolizumab and Vedolizumab Antibodies: Comparison of Commercial Assays with the Vedolizumab Clinical Development Assay.

Vedolizumab (VDZ) is an anti-α 4 β 7 integrin monoclonal antibody approved for inflammatory bowel disease treatment. VDZ serum and antidrug antibody (ADA) concentrations may be used for treatment optimization. In this article, the results of 5 commercial assays (Grifols, Immundiagnostik, Progenika, Sanquin, and Theradiag) measuring VDZ concentration and ADA were compared with those of the reference assays used in VDZ clinical studies. Our findings will assist clinicians in interpreting commercial assay results in the context of VDZ clinical trial data. VDZ-treated patient samples were used to evaluate the agreement between commercial assays and the reference VDZ serum concentration assay, based on linear regression, Bland-Altman, and qualitative agreement analyses. VDZ ADAs were detected using qualitative assays. Specificity, selectivity, accuracy, and precision were assessed using serum samples from healthy donors or patients with IBD (VDZ serum concentration <0.5 mcg/mL) spiked with VDZ, with/without other biologics (identical sample sets per assay). All assays were specific and selective for VDZ. Overall, the commercial assay results for VDZ-spiked samples correlated well with those of the reference serum concentration assay (R 2 ≥ 0.98). Compared with the Immundiagnostik and Theradiag assays, the Grifols, Sanquin, and Progenika assays had the best reference assay agreement (based on regression analysis, Bland-Altman plots, and qualitative agreement [Cohen's kappa ≥0.92]). All immunogenicity assays detected VDZ ADAs; only the reference assay detected VDZ ADAs in the presence of 15 mcg/mL VDZ, advising caution with commercial ADA assays if VDZ is present. All 5 commercial assays are suitable for VDZ therapeutic monitoring and ADA testing. However, the absolute values from the reference assays and the different commercial assays were not comparable, indicating that the same assay must be used for repeated monitoring of VDZ serum concentrations.

Preparation and Characterization of Al-Cu-Al2O3/Gr Nano Compound by Metallic Powder Method and Study of the Effect of the Rolling Process on Physical Properties as Suitable Materials for Heat Sink

Al-Cu-Al2O3 / Gr Nano compound was successfully prepared by metallic powder method, 6 samples were prepared with different weight ratios of graphene, 10% copper and 2.5% Al2O3 and proportions of 0.5, 1, and 1.5 graphene were prepared, the samples were run in a mill from Ceramic in a ratio of 1:5 powder into balls for 35 hours, which helped to break the aluminum and copper particles and reduce their size, which helps in the process of homogeneous mixing of the compound. The samples were compressed at 60 bar pressure and sintered in a vacuum at 550 and 565 degrees Celsius for 60 minutes. Sintering at 550°C proved to be more suitable for the mixture. Two identical sets of samples were prepared. Both SEM were used to investigate the microstructure and components of the sintered nanocomposites. Relative density, hardness, electrical conductivity and thermal conductivity study. The rolling process of the samples was carried out with a percentage of 35% of the sample size successfully at a temperature of 480 degrees Celsius, and this led to an improvement in the density of the samples and hardness and an increase in the diffusion of the reinforces, which led to an improvement in electrical conductivity by 16: 25% and a better improvement in thermal conductivity. Improved up to 1.5% graphene by weight after rolling and 1% graphene before rolling.

Determination of the anisotropic thermal conductivity of an aerogel-based plaster using transient plane source method

Aerogel-based plasters are composite materials with declared thermal conductivities in the range of traditional insulating materials, i.e. 30-50 mW/(m·K). Based on the results from reported field measurements, aerogel-based plasters can significantly reduce the thermal transmittance of uninsulated walls. However, the in-situ measured thermal conductivities have sometimes been higher than the declared values measured in laboratory and in the main direction of the heat flow. Meanwhile, the anisotropic thermal performance of aerogel-based plasters, i.e., deviating thermal performance in the different directions of heat flow, has not been explored yet. The objective of this study is thus to evaluate the anisotropic thermal conductivity of an aerogel-based plaster. This is done in a set of laboratory measurements using the transient plane source method. Six identical and cubic samples with the dimensions of 10×10×10 cm3 were paired two and two, creating three identical sample sets. In total, 360 measurements of thermal conductivity and thermal diffusivity, and 130 measurements for specific heat capacity were conducted. The results indicate a weak anisotropy of less than ±6.5 % between the three directions (x, y, z). Considering the accuracy of the selected measurement technique, better than ±5 %, supplementary measurements using another technique are recommended.

THE INFLUENCE OF INTERRUPTING THE BRAZING PROCESS OF HASTELLOY X SUPERALLOY ON THE PROPERTIES OF THE JOINT

In this paper, disruption of brazing process impact on behavior of filler metal and properties of the joint was investigated. Hastelloy X sheet samples were brazed using Ni-13Cr-4Fe-4Si-2,7B filler alloy. Two identical sets of samples were heated in high vacuum, once load reached specified temperature process was interrupted. Subsequently, one set of samples was tested and second set was subjected to rebrazing in proper time and temperature, dedicated for used filler material. Disruption of the process in following temperature was analyzed: 930, 960, 1050, 1127, 1145, 1170°C. After both processes macroscopic observations of joint, peel test, static tensile test and spreadability analysis were carried out.

Performance characteristics of five immunoassays for SARS-CoV-2: a head-to-head benchmark comparison

SummaryBackgroundSevere acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a global pandemic in 2020. Testing is crucial for mitigating public health and economic effects. Serology is considered key to population-level surveillance and potentially individual-level risk assessment. However, immunoassay performance has not been compared on large, identical sample sets. We aimed to investigate the performance of four high-throughput commercial SARS-CoV-2 antibody immunoassays and a novel 384-well ELISA.MethodsWe did a head-to-head assessment of SARS-CoV-2 IgG assay (Abbott, Chicago, IL, USA), LIAISON SARS-CoV-2 S1/S2 IgG assay (DiaSorin, Saluggia, Italy), Elecsys Anti-SARS-CoV-2 assay (Roche, Basel, Switzerland), SARS-CoV-2 Total assay (Siemens, Munich, Germany), and a novel 384-well ELISA (the Oxford immunoassay). We derived sensitivity and specificity from 976 pre-pandemic blood samples (collected between Sept 4, 2014, and Oct 4, 2016) and 536 blood samples from patients with laboratory-confirmed SARS-CoV-2 infection, collected at least 20 days post symptom onset (collected between Feb 1, 2020, and May 31, 2020). Receiver operating characteristic (ROC) curves were used to assess assay thresholds.FindingsAt the manufacturers' thresholds, for the Abbott assay sensitivity was 92·7% (95% CI 90·2–94·8) and specificity was 99·9% (99·4–100%); for the DiaSorin assay sensitivity was 96·2% (94·2–97·7) and specificity was 98·9% (98·0–99·4); for the Oxford immunoassay sensitivity was 99·1% (97·8–99·7) and specificity was 99·0% (98·1–99·5); for the Roche assay sensitivity was 97·2% (95·4–98·4) and specificity was 99·8% (99·3–100); and for the Siemens assay sensitivity was 98·1% (96·6–99·1) and specificity was 99·9% (99·4–100%). All assays achieved a sensitivity of at least 98% with thresholds optimised to achieve a specificity of at least 98% on samples taken 30 days or more post symptom onset.InterpretationFour commercial, widely available assays and a scalable 384-well ELISA can be used for SARS-CoV-2 serological testing to achieve sensitivity and specificity of at least 98%. The Siemens assay and Oxford immunoassay achieved these metrics without further optimisation. This benchmark study in immunoassay assessment should enable refinements of testing strategies and the best use of serological testing resource to benefit individuals and population health.FundingPublic Health England and UK National Institute for Health Research.

675 Measuring Serum Vedolizumab and Antibodies to Vedolizumab: Comparison of Four Commercially Available Clinical Laboratory Assays to the Assay Used in Vedolizumab Development

INTRODUCTION: Vedolizumab (VDZ), a monoclonal antibody targeting the α4β7 integrin, is approved for the treatment of inflammatory bowel disease (IBD). Treatment optimization with VDZ may be informed by measuring VDZ serum levels. However, the assays used in the GEMINI studies are not commercially available and the relationship to results obtained with commercial assays is unknown. Therefore, VDZ serum level and anti-VDZ antibody (AVA) assays from Grifols Diagnostic Solutions, Immundiagnostik AG, Progenika Biopharma and Theradiag were compared to the assays used by Takeda (Reference; Ref). METHODS: The following identical sample sets were provided for serum level analysis: 1) 10 pooled sample groups (in triplicate) from biologic-naïve IBD patients spiked with up to 70 mg/mL of VDZ, 2) 2 pooled sample groups (in duplicate) biologic-naive subjects, both groups had a combination of 10 mg/mL infliximab (IFX) and 5 mg/mL of adalimumab (ADA), one group with 20 mg/mL VDZ and a second group without VDZ. 3) 23 singlet samples from VDZ-treated IBD patients with free drug concentrations between 8-60 mg/mL. 4) For AVA, 8 singlet samples with AVA (0 - ≥ 0.5 mg/mL), 4 with 15 mg/mL of VDZ and 4 without VDZ. For VDZ-spiked groups 1 and 2, results for replicates were averaged and were compared to the Ref assay results. Slopes were calculated by weighted least squares regression (spiked samples) or by linear regression (patient samples). AVA was detected with a qualitative assay, with results reported as AVA positive or AVA negative. RESULTS: All assays were specific with no cross-reactivity with IFX or ADA and were selective, detecting VDZ in samples that were spiked with all three drugs. Overall Vendor analysis of spiked samples correlated well (R2 ≥ 0.90) with the Ref assay but there were differences in the measured values based on differing slopes (range 0.94-1.56). Likewise, vendor patient sample analysis correlated well with the Ref assay (R2 ≥ 0.95) but slopes differed ((range 0.81-1.71); Figure 1. The Ref AVA assay has a sensitivity of 3.9 ng/mL and detects AVA at 0.5 mg/mL in the presence of VDZ levels of ≥50 mg/mL. All 4 vendor assays detected AVA but did not detect AVA in the presence of 15 mg/mL VDZ. CONCLUSION: Despite good specificity and selectivity, there are differences between the Ref assays Takeda has used in its clinical trials and commercial assays. This should be acknowledged when interpreting assays results with vedolizumab commercial assays.

On the Habitability of Desert Varnish: A Combined Study by Micro-Raman Spectroscopy, X-ray Diffraction, and Methylated Pyrolysis–Gas Chromatography–Mass Spectrometry

In 2020, the ESA ExoMars and NASA Mars 2020 missions will be launched to Mars to search for evidence of past and present life. In preparation for these missions, terrestrial analog samples of rock formations on Mars are studied in detail in order to optimize the scientific information that the analytical instrumentation will return. Desert varnishes are thin mineral coatings found on rocks in arid and semi-arid environments on Earth that are recognized as analog samples. During the formation of desert varnishes (which takes many hundreds of years), organic matter is incorporated, and microorganisms may also play an active role in the formation process. During this study, four complementary analytical techniques proposed for Mars missions (X-ray diffraction [XRD], Raman spectroscopy, elemental analysis, and pyrolysis-gas chromatography-mass spectrometry [Py-GC-MS]) were used to interrogate samples of desert varnish and describe their capacity to sustain life under extreme scenarios. For the first time, both the geochemistry and the organic compounds associated with desert varnish are described with the use of identical sets of samples. XRD and Raman spectroscopy measurements were used to nondestructively interrogate the mineralogy of the samples. In addition, the use of Raman spectroscopy instruments enabled the detection of β-carotene, a highly Raman-active biomarker. The content and the nature of the organic material in the samples were further investigated with elemental analysis and methylated Py-GC-MS, and a bacterial origin was determined to be likely. In the context of planetary exploration, we describe the habitable nature of desert varnish based on the biogeochemical composition of the samples. Possible interference of the geological substrate on the detectability of pyrolysis products is also suggested. Key Words: Desert varnish-Habitability-Raman spectroscopy-Py-GC-MS-XRD-ExoMars-Planetary science. Astrobiology 17, 1123-1137.

ToF-SIMS and Principal Component Analysis Investigation of Denatured, Surface-Adsorbed Antibodies.

Antibody denaturation at solid-liquid interfaces plays an important role in the sensitivity of protein assays such as enzyme-linked immunosorbent assays (ELISAs). Surface immobilized antibodies must maintain their native state, with their antigen binding (Fab) region intact, to capture antigens from biological samples and permit disease detection. In this work, two identical sample sets were prepared with whole antibody IgG, F(ab')2 and Fc fragments, immobilized to either a silicon wafer or a diethylene glycol dimethyl ether plasma polymer surface. Analysis was conducted on one sample set at day 0, and the second sample set after 14 days in vacuum, with time-of-flight secondary ion mass spectrometry (ToF-SIMS) for molecular species representative of denaturation. A 1003 mass fragment peak list was compiled from ToF-SIMS data and compared to a 35 amino acid mass fragment peak list using principal component analysis. Several ToF-SIMS secondary ions, pertaining to disulfide and thiol species, were identified in the 14 day (presumably denatured) samples. A substrate and primary ion independent marker for denaturation (aging) was then produced using a ratio of mass peak intensities according to denaturation ratio: [I61.9534 + I62.9846 + I122.9547 + I84.9609 + I120.9461]/[I30.9979 + I42.9991 + I73.0660 + I147.0780]. The ratio successfully identifies denaturation on both the silicon and plasma polymer substrates and for spectra generated with Mn+, Bi+, and Bi3+ primary ions. We believe this ratio could be employed to as a marker of denaturation of antibodies on a plethora of substrates.

Density improvement of Si quantum dots embedded in Si-rich silicon nitride films by light-filtering rapid thermal processing

Si-rich silicon nitride (SRSN) (SiNx, x ≈ 0.49) films were deposited on Si (100) and quartz substrates by magnetron co-sputtering. For comparison, two sets of identical samples were then treated in a nitrogen atmosphere by conventional rapid thermal processing (CRTP) and light-filtering rapid thermal processing (LRTP) at temperature in a range of 950–1,100 °C, respectively. Raman spectroscopy, grazing incident X-ray diffraction, transmission electron microscope, photoluminescence (PL) and Hall measurements were used to analyze the structure, luminescence, and conductivity of the films. Experimental results show that the samples treated with LRTP posses higher dot number density, crystalline volume fraction, PL intensity and conductivity than the CRTP samples. The quantum effects in rapid thermal processing have a negative influence on the formation and density of Si quantum dots (QDs) in SRSN films. The present work opens new strategy for the formation of high density Si QDs embedded in SRSN films.

Pre-chill with gibberellic acid overcomes seed dormancy of Jerusalem artichoke

Jerusalem artichoke is a valuable source of inulin, a polysaccharide. Inulin is a sustainable source of dietary fiber that enhances the immune system in humans. Although Jerusalem artichoke can be propagated vegetatively, breeders use cross-fertilization to produce novel varieties and hybrids with higher inulin yields. Seed dormancy can hinder the breeding progress because dormancy reduces the number of generations a breeder can obtain in 1 year. Current methods for breaking seed dormancy are time consuming and usually involve removing or pin-pricking the seed coat of these very small seeds, or using several weeks of seed vernalization or several months of storage. Breeders thus require better methods that are less time consuming and achieve higher germination percentages. Here, we studied germination, dormant seed, dead seed, and the seedling growth rate of different Jerusalem artichoke genotypes and seed lots after seeds were treated with different methods to break seed dormancy. We evaluated fresh and stored seed lots using the following treatments: seed samples were planted on top of media moistened with either potassium nitrate, gibberellic acid, or distilled water as control. Two identical sets of samples were planted: one set was germinated in a chamber at alternate 15–25 °C for 28 days, while the second set was pre-chilled at 5 °C for 14 days, before moving the samples to the alternate 15–25 °C chamber for 14 days. Our findings reveal that the highest germination percentage up to 85.3 % was obtained when applying pre-chill with gibberellic acid. This novel dormancy-breaking treatment was thus effective in promoting fresh seed germination.

Energy monitoring and analysis during deformation of bedded-sandstone: Use of acoustic emission

This paper investigates the mechanical behaviour and energy releasing characteristics of bedded-sandstone with bedding layers in different orientations, under uniaxial compression. Cylindrical sandstone specimens (54mm diameter and 108mm height) with bedding layers inclined at angles of 10°, 20°, 35°, 55°, and 83° to the minor principal stress direction, were produced to perform a series of Uniaxial Compressive Strength (UCS) tests. One of the two identical sample sets was fully-saturated with water before testing and the other set was tested under dry conditions. An acoustic emission system was employed in all the testing to monitor the acoustic energy release during the whole deformation process of specimens. From the test results, the critical joint orientation was observed as 55° for both dry and saturated samples and the peak-strength losses due to water were 15.56%, 20.06%, 13.5%, 13.2%, and 13.52% for the bedding orientations 10°, 20°, 35°, 55°, and 83°, respectively. The failure mechanisms for the specimens with bedding layers in 10°, 20° orientations showed splitting type failure, while the specimens with bedding layers in 55°, 83° orientations were failed by sliding along a weaker bedding layer. The failure mechanism for the specimens with bedding layers in 35° orientation showed a mixed failure mode of both splitting and sliding types. Analysis of the acoustic energy, captured from the acoustic emission detection system, revealed that the acoustic energy release is considerably higher in dry specimens than that of the saturated specimens at any bedding orientation. In addition, higher energy release was observed for specimens with bedding layers oriented in shallow angles (which were undergoing splitting type failures), whereas specimens with steeply oriented bedding layers (which were undergoing sliding type failures) showed a comparatively less energy release under both dry and saturated conditions. Moreover, a considerable amount of energy dissipation before the ultimate failure was observed for specimens with bedding layers oriented in shallow angles under both dry and saturated conditions. These results confirm that when rock having bedding layers inclined in shallow angles the failures could be more violent and devastative than the failures of rock with steeply oriented bedding layers.

Comparison of candidate vCJD in vitro diagnostic assays using identical sample sets

With four transfusion related transmissions of variant Creutzfeldt-Jakob Disease (vCJD), three of which developed clinical disease and the other died of other causes but was positive for markers of infection, there is an increased urgency to identify and implement a test for blood donor screening. With limited amounts of blood samples from vCJD cases available test evaluation is challenging. Alternative approaches are therefore needed. Control and vCJD tissues homogenates, where levels of markers of infectivity are known, were sequentially diluted in pooled human plasma. Identical sets of samples were provided blind to research groups developing diagnostic tests for vCJD; identical sample sets allows for direct comparisons of sensitivity to be made. Control and vCJD tissue homogenates were sequentially diluted in pooled human plasma (detergent solvent treated or cryo-depleted) supplied by commercial fractionators. Dilutions of vCJD tissues were within and beyond the limits of detection previously determined by the conformation-dependent immunoassay (Cooper et al.: Vox Sang 2007;92:302-310; Bellon et al.: J Gen Virol 2003;84: 1921-1925). A number of methods were used for the analysis of the blinded panels; with background signal from the normal prion protein (PrP) being removed by digestion with proteinase, epitope protection or selective capture of PrP(tse). Assay sensitivities were directly compared using identical sample sets. This approach identified several transmissible spongiform encephalopathies (TSE) diagnostic tests, based on different principles, high in analytical sensitivity that reproducibly detected markers of vCJD infectivity in tissue homogenates. The approach outlined has successfully compared in vitro diagnostics assays for their sensitivity and reproducibility and is a first step toward the evaluation of an assay suitable for blood donor screening/diagnosis of vCJD.

3-Hydroxypropyl Mercapturic Acid Interlaboratory Method Standardization to Measure Acrolein Exposure in Human Urine

PP-29-059 Background/Aims: An interlaboratory comparison was set up to measure the consistency of analysis of 3-HPMA (3-hydroxypropyl mercapturic acid), a biomarker of acrolein exposure, across 4 participating laboratories using LC-MS/MS. In the first phase, measurement differences between the laboratories were observed, and it was concluded that further method of standardization was required. A number of potential sources of variance, including differences in calibration standards used, were identified. Methods: Following a review of the 3-HPMA methods used by each laboratory, a second interlaboratory comparison study was initiated incorporating a common internal standard in the operating protocol (3-HPMA-13C3-15N). The objective of the new study was to validate 3-HPMA measurement capability by LC-MS/MS between the same 4 laboratories, using identical sets of samples and the same internal standard. Results: The results from this second interlaboratory study demonstrate that the use of a common internal standard improved the reproducibility of 3-HPMA quantification by lowering the average interlaboratory coefficient of variation (CoV) from >20% to 13%. This compares favorably with World Health Organization standardized clinical methods, which typically report average interlaboratory CoV above 10% (1–2). As expected, the repeatability of the quantification was not adversely affected by the new internal standard, given that no other changes were made to the overall procedures within the individual laboratories. Conclusion: In conclusion, the results from this new study demonstrate that the use of a common internal standard improved the interlaboratory reproducibility of 3-HPMA quantification between participating laboratories.

Further Studies on Reciprocity Effects for Accelerated Ozone Tests Compared to Ambient Air Exposure

The digital print industry is working toward the goal of adopting improved test methods for image permanence. This study focused on the impact of ozone concentration on the performance of inkjet print samples. Results from accelerated tests were compared to data collected from identical sample sets exposed to ambient air and to filtered air. Reciprocity failure was observed for some samples in the test, but the results also verified earlier work in which ambient air exposure can be generally linked to accelerated ozone test results. This research is part of ongoing work contributing to the development of standardized test methods for image permanence.

Concordant Proficiency in Measurement of T-Cell Immunity in Human Immunodeficiency Virus Vaccine Clinical Trials by Peripheral Blood Mononuclear Cell and Enzyme-Linked Immunospot Assays in Laboratories from Three Continents

The gamma interferon (IFN-gamma) enzyme-linked immunospot (ELISPOT) assay is used routinely to evaluate the potency of human immunodeficiency virus (HIV) vaccine candidates and other vaccine candidates. In order to compare candidates and pool data from multiple trial laboratories, validated standardized methods must be applied across laboratories. Proficiency panels are a key part of a comprehensive quality assurance program to monitor inter- and intralaboratory performance, as well as assay performance, over time. Seven International AIDS Vaccine Initiative-sponsored trial sites participated in the proficiency panels described in this study. At each laboratory, two operators independently processed identical sample sets consisting of frozen peripheral blood mononuclear cell (PBMC) samples from different donors by using four blind stimuli. PBMC recovery and viability after overnight resting and the IFN-gamma ELISPOT assay performance were assessed. All sites demonstrated good performance in PBMC thawing and resting, with a median recovery of 78% and median viability of 95%. The laboratories were able to detect similar antigen-specific T-cell responses, ranging from 50 to >3,000 spot-forming cells per million PBMC. An approximate range of a half log in results from operators within or across sites was seen in comparisons of antigen-specific responses. Consistently low background responses were seen in all laboratories. The results of these proficiency panels demonstrate the ability of seven laboratories, located across three continents, to process PBMC samples and to rank volunteers with differential magnitudes of IFN-gamma ELISPOT responses. These findings also illustrate the ability to standardize the IFN-gamma ELISPOT assay across multiple laboratories when common training methods, reagents such as fetal calf serum, and standard operating procedures are adopted. These results are encouraging for laboratories that are using cell-based immunology assays to test HIV vaccines and other vaccines.

Measuring the upset of CMOS and TTL due to HPM-signals

Abstract. To measure the performance of electronic components when stressed by High Power Microwave signals a setup was designed and tested which allows a well-defined voltage signal to enter the component during normal operation, and to discriminate its effect on the component. The microwave signal is fed to the outside conductor of a coaxial cable and couples into the inner signal line connected to the device under test (DUT). The disturbing HF-signal is transferred almost independent from frequency to maintain the pulse shape in the time domain. The configuration designed to perform a TEM-coupling within a 50 Ohm system prevents the secondary system from feeding back to the primary system and, due to the geometrical parameters chosen, the coupling efficiency is as high as 50–90%. Linear dimensions and terminations applied allow for pulses up to a width of 12ns and up to a voltage level of 4–5 kV on the outside conductor. These pulse parameters proved to be sufficient to upset the DUTs tested so far. In more than 400 measurements a rectangular pulse of increasing voltage level was applied to different types of CMOS and TTL until the individual DUT was damaged. As well the pulse width (3, 6 or 12 ns) and its polarity were varied in single-shot or repetitive-shot experiments (500 shots per voltage at a repetition rate of 3 Hz). The state of the DUT was continuously monitored by measuring both the current of the DUT circuit and that of the oscillator providing the operating signal for the DUT. The results show a very good reproducibility within a set of identical samples, remarkable differences between manufacturers and lower thresholds for repetitive testing, which indicates a memory effect of the DUT to exist for voltage levels significantly below the single-shot threshold.

The Influence of Various Hematology Analyzers on Component Platelet Counts

Hematology analyzers designed to count platelets in samples of whole blood are used to enumerate the total number of platelets in components prepared for transfusion. This report addresses the issue of variability in platelet counts obtained with different models of hematology analyzers. The influence of a common calibration procedure, involving one level of porcine platelets, on the extent of variability was also evaluated. Identical sets of samples of simulated and apheresis-derived human platelets were counted by multiple laboratories in 3 separate studies. In the first 2 exercises, 7 samples of both porcine platelets and modified goat erythrocytes with targeted platelets counts from 0.2 to 4.0 x 10(12)/L were counted without prior dilution. In both exercises, the samples were counted multiple times after routine calibration using instructions provided by the manufacturers of the various hematology analyzers used. In the second exercise, the samples were recounted after the hematology analyzers were recalibrated with a common calibrant consisting of porcine platelets at a targeted concentration of 0.5 x 10(12)/L. In the first and second exercises, 20 and 18 hematology analyzers were used, respectively. In the third exercise, 6 samples prepared from a single unit of apheresis platelets with targeted counts from 0.2 to 1.64 x 10(12)/L were shipped by an overnight courier and counted in triplicate on the day of arrival. Eleven hematology analyzers were used. The influence of recalibration was evaluated statistically by using the 95% prediction interval for the mean of a future set of observations. The platelet counts measured with a specific type of hematology analyzer provided the data to calculate the 95% prediction interval. With routine calibration, a wide variability in platelet counts was observed with all levels of both simulated and apheresis-derived human platelets. For example, with porcine platelets at a targeted level of 0.4 x 10 (12)/L, the platelet counts ranged from 0.31 to 0.47 x 10(12)/L. Recalibration reduced the extent of variability observed with all levels of simulated and apheresis-derived human platelets by increasing the observed platelet counts determined with a subset of hematology analyzers that produced platelet counts in the lower portion of the range. With recalibration, the mean platelet counts obtained with most hematology analyzers, especially with samples having targeted platelet levels no greater than 1.0 x 10(12)/L, were within or near the 95% prediction interval determined with the instruments that provided the highest platelet counts with routine calibration. With recalibration, the reproducibility of the platelet counts was considered to be good for all hematology analyzers with all levels of simulated and apheresis-derived human platelets for most of the instruments. The coefficient of variance did not exceed 6%, with most of the values ranging from 1% to 3%. This study therefore found that the platelet counts of platelet concentrates can be markedly influenced by the type of hematology analyzer used. A common calibration procedure designed specifically for the range of platelet counts in platelet products may be beneficial considering that many different hematology analyzers are being used to count platelets.

Comparison of genotypic-based microbial source tracking methods requiring a host origin database

Microbial source tracking (MST) results, obtained using identical sample sets and pulsed field gel electrophoresis (PFGE), repetitive element PCR (rep-PCR) and ribotyping techniques were compared. These methods were performed by six investigators in analysis of duplicate, blind sets of water samples spiked with feces from five possible sources (sewage, human, dog, cow and seagull). Investigators were provided with samples of the fecal material used to inoculate the water samples for host origin database construction. All methods correctly identified the dominant source in the majority of the samples. Modifications of some of these methods correctly identified the dominant sources in over 90% of the samples; however, false positive rates were as high as 57%. The high false positive rates appeared to be indirectly proportional to the levels of stringency applied in pattern analysis. All the methods produced useful data but the results highlighted the need to modify and optimize these methods in order to minimize sources of error.

Inhibition in a microgravity environment of the recovery of Escherichia coli cells damaged by heavy ion beams during the NASDA ISS phase I program of NASA Shuttle/Mir mission no. 6.

We participated in a space experiment, part of the National Space Development Agency of Japan (NASDA) Phase I Space Radiation Environment Measurement Program, conducted during the National Aeronautics and Space Administration (NASA) Shuttle/Mir Mission No. 6 (S/MM-6) project. The aim of our study was to investigate the effects of microgravity on the DNA repair processes of living organisms in the <Realtime Radiation Monitoring Device III (RRMD III)> in orbit. Heavy ion beam radiation- or ç-irradiation-damaged biological samples of Escherichia coli and the radioresistant bacterium Deinococcus radiodurans were prepared and placed in a biospecimen box, which was loaded into the RRMD III sensor unit of the Space Shuttle. Two identical sets of samples were left in the Spacehab's Payload Processing Facility (SPPF) in Florida, USA, as a control. (flight No. STS-84) was launched from NASA John F. Kennedy Space Center (KSC) in Florida, USA, on May 15, 1997. The mission duration was 9.22 days. An astronaut activated the biological samples in the biospecimen box in the Spacehab during orbit in order to start repair of the DNA damaged by heavy ion beams or ç-irradiation and the samples were incubated for 19 h 35 min at about 22ûC, the cabin temperature. The control specimens in the SPPF were subjected to the same treatment under terrestrial gravity. After returned to earth, we investigated cell recovery by comparing the repair of the radiation-damaged DNA of E. coli and D. radiodurans in the microgravity environment in space with that on Earth. The results indicated that the DNA repair process of E. coli, but not of D. radiodurans, cells was inhibited in a microgravity environment.