Common Reference Material Research Articles

A Novel High-Throughput Immunoaffinity LC-MS/MS Assay for P-III-NP and Other Fragments of Type III Procollagen in Human Serum.

The amino-terminal propeptide of type III procollagen (P-III-NP) is used with IGF-I to detect the illicit use of growth hormone and to monitor growth hormone therapy. However, the only currently available assays for P-III-NP are immunoassays, which are not well harmonized. In addition, other fragments of type III procollagen may better evaluate collagen turnover. We aimed to develop a high-throughput assay using immunoaffinity enrichment coupled to ultra-high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) to quantify peptides belonging to three different regions of type III procollagen in human serum simultaneously. To facilitate higher throughput, we transferred the assay from microcentrifuge tubes to a 96-well plate format with partially automated pipetting. The method was linear (Pearson's R ≥ 0.994) over an estimated concentration range of 1.35-13.3 nM, 0.04-2.28 nM, and 0.26-5.1 nM for each surrogate peptide of P-III-NP, collagen degradation products, and the carboxyl-terminal propeptide, respectively. Intra-day and inter-day imprecision were both < 13.6%, and the results of robustness testing were also encouraging. The method was successfully applied to capillary blood samples obtained using Tasso+ microsampling devices. Modest correlation of P-III-NP concentration was observed between our new method and a WADA-approved immunoassay (N = 40, Pearson's R = 0.789) with a significant bias of -87.8%. Our method simultaneously quantifies four peptides belonging to three regions of type III procollagen in human serum. High bias between assays highlights the need for common higher-order calibrators or reference materials to help improve the comparability of results across laboratories.

Discrepancies between two total IgE assays and difference in reference intervals in healthy adults

ObjectiveTo compare total immunoglobulin (Ig) E assay performance characteristics between Abbott Architect and Siemens Immulite test systems. Reference intervals were also determined for both platforms in an American population of healthy adults. MethodsAgreement of the two total IgE assays was evaluated in a cohort of 331 subjects with normal complete blood count (CBC) and comprehensive metabolic panel (CMP) results. Reference intervals were established in 302 subjects after exclusion of atopic individuals on the Abbott Architect and Siemens Immulite test systems. ResultsWe demonstrated a 32% positive bias for total IgE quantitation on the Siemens Immulite platform compared to the Abbott Architect, despite both methods calibrated against the same WHO international reference material (75/502), Furthermore, the upper limit of the reference interval (95th percentile) was determined to be higher for the Siemens Immulite assay compared to the Abbott Architect (132 and 102 IU/mL, respectively). ConclusionDespite the use of a common WHO reference material for total IgE assay calibration, significant differences in quantitation was observed between two FDA-cleared test systems. Given that, it is warranted for clinical laboratories to verify vendor established reference intervals and adjust accordingly based on internal assessment of the normal range.

Mild selective photochemical oxidation of an organic sulfide using OxP-polyimide porous polymers as singlet oxygen generators

ABSTRACT A series of porous organic polymers based on a singlet oxygen generating oxoporphyinogen (‘OxP’) has been successfully prepared from a pseudotetrahedral OxP-tetraamine precursor (OxP(4-NH 2 Bn) 4 ) by its reaction with tetracarboxylic acid dianhydrides under suitable conditions. Of the compounds studied, those containing naphthalene (OxP-N) and perylene (OxP-P) spacers, respectively, have large surface areas (~530 m2 g−1). On the other hand, the derivative with a simple benzene spacer (OxP-B) exhibits the best 1O2 generating capability. Although the starting OxP-tetraamine precursor is a poor 1O2 generator, its incorporation into OxP POPs leads to a significant enhancement of 1O2 productivity, which is largely due to the transformation of NH2 groups to electron-withdrawing diimides. Overall 1O2 production efficacy of OxP-POPs under irradiation by visible light is significantly improved over the common reference material PCN-222. All the materials OxP-B, OxP-N and OxP-P promote oxidation of thioanisole involving conversion of ambient triplet state oxygen to singlet oxygen under visible light irradiation and its reaction with the sulfide. Although the reaction rate of the oxidation promoted by OxP POPs is generally lower than for conventional materials (such as PCN-222) or previously studied OxP derivatives, undesired overoxidation of the substrate to methyl phenyl sulfone is suppressed. For organic sulfides, selectivity of oxidation is especially important for detoxification of mustard gas (bis(2-chloroethyl)sulfide) or similarly toxic compounds since controlled oxidation leads to the low toxicity bis(2-chloroethyl)sulfoxide while overoxidation leads to intoxification (since bis(2-chloroethyl)sulfone presents greater toxicity to humans than the sulfide substrate). Therefore, OxP POPs capable of promoting selective oxidation of sulfides to sulfoxides have excellent potential to be used as mild and selective detoxification agents.

Multi-omics data integration using ratio-based quantitative profiling with Quartet reference materials

Characterization and integration of the genome, epigenome, transcriptome, proteome and metabolome of different datasets is difficult owing to a lack of ground truth. Here we develop and characterize suites of publicly available multi-omics reference materials of matched DNA, RNA, protein and metabolites derived from immortalized cell lines from a family quartet of parents and monozygotic twin daughters. These references provide built-in truth defined by relationships among the family members and the information flow from DNA to RNA to protein. We demonstrate how using a ratio-based profiling approach that scales the absolute feature values of a study sample relative to those of a concurrently measured common reference sample produces reproducible and comparable data suitable for integration across batches, labs, platforms and omics types. Our study identifies reference-free ‘absolute’ feature quantification as the root cause of irreproducibility in multi-omics measurement and data integration and establishes the advantages of ratio-based multi-omics profiling with common reference materials.

Interlaboratory variation for NT-proBNP among Swedish laboratories in an external quality program 2011-2021.

NT-proBNP is frequently used for ruling out heart failure. Different cut-offs are used depending on the clinical context, e.g. an acute or chronic condition. Medical decision limits have been suggested at 125 and 300ng/L or 400ng/L in international guidelines. However, there is limited standardization between NT-proBNP methods and using the same blood sample might cause different treatment of patients. Data from the external quality assessment program for NT-proBNP from Equalis, Sweden, were extracted for the period 2011-2021, and categorized according to manufacturer. Manufacturer median NT-proBNP values were compared to total median values. CV% was calculated for each manufacturer and in comparison to different levels of NT-proBNP. Roche was the most common method, and its median results were closest to the median consensus results. When looking at the total CV at NT-proBNP levels in the range of 0-500ng/L, the total CV varied from 4 to 27%. During 2019-2021, Siemens (Immulite, Centaur, Atellica) yielded results 16-20% above the consensus median depending on sample level. Similarly, Abbott was 5-7% above, while Roche and Siemens Stratus were 1% respectively 6-10% below the consensus median. The introduction of new manufacturers and methods in 2017 have caused the agreement between manufacturers to decline. This highlights the need for a common calibrator and reference materials, particularly since medical decision limits in guidelines, e.g. European Society of Cardiology 2021, which are mostly based on Roche methods, do not take these method differences into account.

Quantifying Long‐Term Reproducibility of Zircon Reference Materials by U‐Pb LA‐ICP‐MS Dating

Inter‐session excess variance of U/Pb and Pb/Pb ratios in LA‐ICP‐MS zircon dating is the largest contributor to systematic errors, which in turn limit the accuracy of age determinations. Quantifying long‐term excess variance of reference materials allows for the estimation of excess variance in samples, but such compilations are not available in the literature. Here, we present the results of over 100 measurement sessions over 6 years for seven common zircon reference materials and calculate a characteristic excess variance for each. For 206Pb/238U ages, these values (2s) are as follows: AusZ7‐1 = 1.7% (115 sessions), AusZ7‐5 = 0% (74), OD‐3 = 0.6% (19), Temora2 = 1.2% (86), Plešovice = 1.1% (100), 91500 = 1.0% (146) and Mud Tank = 3.0% (12). For 207Pb/206Pb ages, smaller excess variances are observed: Temora2 = 0.3%, Plešovice = 0.4%, Mud Tank = 1.7% and 91500 = 0.4%. These values are well‐constrained estimates of inter‐session excess variance for the ETH Zürich LA‐ICP‐MS laboratory using a sector‐field ICP‐MS, and may provide either a first‐order estimate for other laboratories or a value to compare against their own reference material compilations.

Effectiveness of biochar filters vegetated with Echinochloa pyramidalis in domestic wastewater treatment.

The use of biochar in constructed wetlands for domestic wastewater treatment is gradually being acclaimed by environmentalists due to its high specific surface area and porosity. In this study, the effectiveness of corn cob biochar (CCB) and rice husk biochar (RHB) in vertical flow constructed wetlands vegetated with Echinochloa pyramidalis was studied with sand as common reference material. The filters were fed with primarily treated domestic wastewater at a hydraulic loading rate of about 350 L/m2/day for 6 months. Water samples were collected monthly for physicochemical and bacteriological analysis and plant growth assessed every two weeks throughout the study. Biochar filters were highly performant in wastewater improvement with no significant differences between the biochar types. Both biochars were more efficient than sand in the removal of chemical oxygen demand (COD), biochemical oxygen demand (BOD), true colour, total suspended solids (TSS) and total dissolved solids (TDS). However, sand filters performed better in the reduction of nutrients. All wetlands showed positive plant growth though the plants did not significantly affect the performance of the different filters for most parameters. However, a better plant growth was observed in the CCB filters. The study shows that CCB and RHB can effectively replace sand as substrates in constructed wetlands for wastewater treatment.

Final report on CCQM-K167: carbon isotope delta measurements of vanillin

Main textThe CCQM Isotope Ratio Working Group (IRWG) determined that an additional key comparison of carbon isotope delta measurements was required to capture the progress of this field. Vials containing 0.25 mg of vanillin were prepared at NRC, and evaluated for bottle-to-bottle homogeneity prior to distribution to the eight participating institutes. Participants were able to choose any suitable method and reference materials for carbon isotope delta measurements, and report a carbon isotope delta value and the associated uncertainty, and analysis details. To determine the key comparison reference value (KCRV) and its associated uncertainty, the NRC in collaboration with the NIST, developed a multivariate Bayesian random laboratory effects model, which also incorporates the uncertainty due to bottle-to-bottle homogeneity and any correlations between the reported results that arise due to the use of common reference materials. The KCRV for this study was determined to be -25.833 ‰ relative to the VPDB, with associated uncertainty of 0.028 ‰, and expanded uncertainty of 0.056 ‰ (k=2). All the results reported by the participants in this study were considered equivalent to the KCRV.To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database https://www.bipm.org/kcdb/.The final report has been peer-reviewed and approved for publication by the CCQM, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

The VitMin Lab Sandwich-ELISA Assays for Iron and Inflammation Markers Compared Well with Clinical Analyzer Reference-Type Assays in Subsamples of the Nepal National Micronutrient Status Survey

BackgroundThe low cost and small specimen volume of the VitMin Lab ELISA assays for serum ferritin (Fer), soluble transferrin receptor (sTfR), C-reactive protein (CRP), and α-1-acid glycoprotein (AGP) have allowed their application to micronutrient surveys conducted in low-resource countries for ∼2 decades. ObjectivesWe conducted a comparison between the ELISA and reference-type assays used in the US NHANES. MethodsUsing the Roche clinical analyzer as a reference, we measured random subsets of the 2016 Nepal National Micronutrient Status Survey (200 serum samples from children aged 6–59 mo; 100 serum samples from nonpregnant women) for Fer, sTfR, CRP, and AGP. We compared the combined data sets with the ELISA survey results using descriptive analyses. ResultsThe Lin's concordance coefficients between the 2 assays were ≥0.89 except for sTfR (Lin's ρ = 0.58). The median relative difference to the reference was as follows: Fer, −8.5%; sTfR, 71.2%; CRP, −19.5%; and AGP, −8.2%. The percentage of VitMin samples agreeing within ±30% of the reference was as follows: Fer, 88.5%; sTfR, 1.70%; CRP, 74.9%; and AGP, 92.9%. The prevalence of abnormal results was comparable between the 2 assays for Fer, CRP, and AGP, and for sTfR after adjusting to the Roche assay. Continued biannual performance (2007–2019) of the VitMin assays in CDC's external quality assessment program (6 samples/y) demonstrated generally acceptable performance. ConclusionsUsing samples from the Nepal survey, the VitMin ELISA assays produced mostly comparable results to the Roche reference-type assays for Fer, CRP, and AGP. The lack of sTfR assay standardization to a common reference material explains the large systematic difference observed for sTfR, which could be corrected by an adjustment equation pending further validation. This snapshot comparison together with the long-term external quality assessment links the survey data generated by the VitMin Lab to the Roche assays used in NHANES.

Precise analysis of antimony isotopic composition in geochemical materials by MC-ICP-MS

Antimony (Sb) isotopes have been shown to be promising tracers for studying Sb cycling in the environment and its impact on ecosystem and human health. Yet precise measurements of Sb isotopic composition have been challenged by low Sb concentrations and high matrix effect and by the lack of a common reference material (zero-delta). Here we report an improved analytical scheme that is capable of high-precision measurement of Sb isotopes in environmental and geochemical samples. The method employs a two-column ion exchange set-up to remove interfering matrix elements to allow Sb isotopic measurements with low Sb (< 0.15 μg/g) and high matrix content. The instrument mass bias is corrected by combining the sample-standard bracketing method with internal normalization of cadmium. To allow for interlaboratory comparisons, a standard reference material certified for the total Sb concentration, NIST SRM 3102a, is recommended as the reference standard (“zero-delta”) for Sb isotope measurements. The improved method is applied for high-precision and high-accuracy Sb isotope composition measurements in environmental and geochemical reference samples with a minimum Sb required per analysis as low as 3.0 ng. By expressing our data and previously reported literature data against the same reference standard (NIST SRM 3102a), we provide an update on isotopic compositions of various types of environmental, geological and anthropogenic materials. The improved analytical method and database aid further studies on the mechanisms of Sb isotope fractionation and its application as a tracer for the study of the sources, processes and fate of Sb in the environment.

Review of techniques, challenges, and new developments for calcium isotope ratio measurements

Calcium isotope meaurements apply to problems spanning the ‘cosmos to benthos.’ Fractionation, source mixing, radioactive decay, and nucleosynthetic processes cause variations in Ca isotope ratios, which can be measured using different types of mass spectrometers. Early measurements employed thermal ionization mass spectrometry (TIMS), which continues to be the most common and precise reference technique. An increasing number of studies use multi-collector inductively coupled plasma mass spectrometry (MC-ICPMS). Both TIMS and MC-ICPMS can yield high-quality data for δ44/40Ca and δ44/42Ca, respectively, but wide ranges in precision exist. Sample size requirements are lower for TIMS, but MC-ICPMS offers faster throughput. Both techniques require purifying Ca by ion-exchange chromatography when analyzing samples with complex matrices. In-situ techniques, like laser ablation MC-ICPMS (LA-MC-ICPMS) and secondary ion mass spectrometry (SIMS), do not require purification. In-situ techniques provide high spatial resolution, but LA-MC-ICPMS and (nano)-SIMS suffer from relatively poor precision compared to TIMS and MC-ICPMS, and the methods lack well-characterized standards, which are essential for comparing datasets.The past few years have seen advances in Ca purification techniques, including automated methods that accelerate sample throughput. The next generation of Ca purification techniques should focus on complete purification and quantitative separation of low-Ca samples. Application of collision-cell technology for suppressing Ar-related molecular isobaric interferences will improve quantification of 40Ca using certain MC-ICPMS models. Similarly, development of new amplifier technology will foster high-precision Faraday cup measurements of low abundance Ca isotopes. Adopting common reference materials, including newly developed standards with complex matrices, would signficantly aid inter-laboratory comparisons, method development, and data compilation efforts. In general, at least two widely-used standards should be measured along with unknown samples to confirm data quality, as well as improve utilization of published Ca isotope datasets by a broader community.

SPX Flow Inc, USA

Antimony (Sb) isotopes have been shown to be promising tracers for studying Sb cycling in the environment and its impact on ecosystem and human health. Yet precise measurements of Sb isotopic composition have been challenged by low Sb concentrations and high matrix effect and by the lack of a common reference material (zero-delta). Here we report an improved analytical scheme that is capable of high-precision measurement of Sb isotopes in environmental and geochemical samples. The method employs a two-column ion exchange set-up to remove interfering matrix elements to allow Sb isotopic measurements with low Sb (< 0.15 μg/g) and high matrix content. The instrument mass bias is corrected by combining the sample-standard bracketing method with internal normalization of cadmium. To allow for interlaboratory comparisons, a standard reference material certified for the total Sb concentration, NIST SRM 3102a, is recommended as the reference standard (“zero-delta”) for Sb isotope measurements. The improved method is applied for high-precision and high-accuracy Sb isotope composition measurements in environmental and geochemical reference samples with a minimum Sb required per analysis as low as 3.0 ng. By expressing our data and previously reported literature data against the same reference standard (NIST SRM 3102a), we provide an update on isotopic compositions of various types of environmental, geological and anthropogenic materials. The improved analytical method and database aid further studies on the mechanisms of Sb isotope fractionation and its application as a tracer for the study of the sources, processes and fate of Sb in the environment.

Measurement harmonization and traceability for trace element analyses across the Children's Health Exposure Analysis Resource laboratory network

Harmonization and traceability are related metrological principles that are indispensable to assuring measurement comparability across different biomonitoring studies. The Children's Health Exposure Analysis Resource (CHEAR) was established in 2015 with six laboratories providing environmental exposure measurements on biospecimens. To ensure harmonization across studies for trace elements, CHEAR used a multi-faceted approach that included: 1) an initial interlaboratory validation exercise based on the analysis of certified blood and urine reference materials; 2) frequent participation in an established interlaboratory proficiency program for trace elements; and 3) analysis of a common pool of well-characterized biological reference materials with each analytical batch. Method accuracy and precision were established for each laboratory via analysis of NIST SRM 955c Toxic Elements in Caprine Blood, SRM 2668 Toxic Elements in Frozen Human Urine and SRM 3668 Mercury, Perchlorate, and Iodide in Frozen Human Urine. The differences among the six laboratories for As, Cd, Hg, and Mn in urine and Cd, Hg, and Pb in whole blood were judged to be fit-for-purpose. Interlaboratory performance over a 5-year period demonstrated an improvement in performance, such that for 2018–2019, >99% of challenges for urine As, Cd, Hg, and Mn, and 95% for whole blood Cd, Hg, Pb, and Mn, were found to be satisfactory. The CHEAR common reference materials were analyzed by at least 5 laboratories for 22 elements in urine and 13–14 elements in whole blood, thus providing a rich source of data to assess intra- and inter-run performance. The suite of trace elements with assigned values in both blood and urine matrices are more comprehensive than similar reference materials from other sources, and is reflective of the concentrations necessary to support biomonitoring studies. While some areas for future improvement were identified, significant progress was made to improve harmonization of trace element measurements in biological matrices among the CHEAR network labs.

Slow sand filtration of raw wastewater using biochar as an alternative filtration media

The efficiency of anaerobic biofilters (AnBF) as low-cost wastewater treatment systems was investigated. Miscanthus-biochar was used as filtration media and compared with sand as a common reference material. Raw sewage from a municipal wastewater treatment plant was stored in a sedimentation tank for two days to allow pre-settlement of wastewater particles. Subsequently, wastewater was treated by AnBFs at 22 °C room temperature at a hydraulic loading rate of 0.05 m∙h−1 with an empty bed contact time of 14.4 h and a mean organic loading rate of 509 ± 173 gCOD∙m−3∙d−1. Mean removal of chemical oxygen demand (COD) of biochar filters was with 74 ± 18% significantly higher than of sand filters (61 ± 12%). In contrast to sand filters with a mean reduction of 1.18 ± 0.31 log-units, E. coli removal through biochar was with 1.35 ± 0.27 log-units significantly higher and increased with experimental time. Main removal took place within the schmutzdecke, a biologically active dirt layer that develops simultaneously on the surface of filter beds. Since the E. coli contamination of both filter materials was equal, the higher removal efficiency of biochar filters is probably a result of an improved biodegradation within deeper zones of the filter bed. Overall, performance of biochar filters was better or equal compared to sand and have thus demonstrated the suitability of Miscanthus-biochar as filter media for wastewater treatment.

High-Sensitivity Measurement of Cr Isotopes by Double Spike MC-ICP-MS at the 10 ng Level.

High-sensitivity and high-precision (2 SD ≤ 0.06‰) measurement of chromium (Cr) isotopes at the 10 ng level was successfully carried out using double spike multiple-collector inductively couple plasma mass spectrometry (MC-ICP-MS). To enhance the signal sensitivity and stability, the Aridus II desolvating nebulizer system was improved by placing its waste gas trap bottle in an ice chamber (5 °C cold trap). This setup, beyond Cr isotope analysis, can be applied to most heavy metal isotope measurements. The sensitivity of the 52Cr signal is ≥300 V mg-1 L (with a 1011Ω amplifier and a 110 μL min-1 uptake rate), an enhancement of ≥1.5 times compared to the Aridus II without the cold trap. In addition, the relative standard deviation of the 52Cr signal varied ≤4% over 8 h, demonstrating high stability. The δ53Cr values of common geological reference materials determined using 10 ng of Cr are in excellent agreement with results measured at 25 ng and 50 ng and are consistent with previous determinations, validating the accurate and precise Cr isotope ratio measurements. An empirical method is proposed to correct for the residual (after subtraction) effect of Fe interference on δ53Cr determination. This method relies on a linear relationship between the [Fe]/[Cr] and δ53Cr shift within one analytical session. Finally, we report the δ53Cr values of 19 new reference materials, ranging from -0.44‰ to 0.49‰. Among them, GSS-7 (-0.44 ± 0.02‰, 2 SD, n = 5), GSS-4 (0.48 ± 0.02‰, 2 SD, n = 5), and GSD-10 (0.49 ± 0.05‰, 2 SD, n = 5) can be used as candidate reference materials for interlaboratory comparisons to complement existing ones that are mostly isotopically unfractionated from the bulk silicate earth.

Abstract 438: Multicenter evaluation of circulating tumor DNA assays

Abstract Measurements of circulating tumor DNA (ctDNA) hold great potential to detect residual disease, therapeutic response monitoring and tumor evolution. However, there are many technical challenges including trace levels of circulating cell free DNA 1-10 ng/ml in plasma (300-3,000 genome equivalents/ml), DNA fragment size 170 bps found in the circulation, low variant allelic fractions ~ 0.1% and the need to detect all variant types SNVs, insertions, deletions, CNVs and structural variants. CANCER-ID partners comprise a consortium of experts, companies and institutions for blood -based biomarker validation, assay development, clinical sciences and bioinformatics. This CANCER-ID ring trial was established to help standardize complex ctDNA diagnostic assays by testing a highly complex reference material. The trial looks to improve concordance and to evaluate analytical performance by collecting metrics covering the entire workflow from sample preparation through variant reporting. The common reference material, Seraseq ctDNA Complete, used in the trial is human genomic DNA background material from the Genome-in-a-bottle consortium mixed with synthetic DNA sized to ~170 bp and formulated in artificial plasma. The material contains 25 variants per sample including 12 SNVs (8 genes), 5 deletions (3 genes), 2 insertions (2 genes), 3 CNVs (3 genes) and 3 fusion pairs (3 genes). The variant allelic fractions range from 0.1% to 5% for the SNVs, INDELS and ~ 1 -6 additional copy number for ERBB2, MET and MYC genes. Interlab and intralab testing was done on Roche AVENIO targeted Assay, Agilent SureSelect XT custom assay, QiaSeq Targeted DNA panel, Qiagen GeneRead QIAAct Lung Assay, one clinically validated digital droplet assay and one research digital droplet PCR assays and Agena Bioscience’s UltraSEEK Lung Panel. Data analysis, peer comparison and reporting of common and unique QC metrics was accomplished through the cloud based, analytical QC software iQ NGS QC Management. The results demonstrate that a common full process reference material can be used to compare distinctly different assays formats at different levels of input DNA by mutations. For the 1% AF material, the BRAF V600 variant was detected by all assay formats and the cv’s ranged from 12%-52% or on average for all variants measured by the assay ranged from 11% to 46%. The study furthermore shows that peer review analytical QC software is useful in establishing standardization of ctDNA assays. This work is supported by IMI JU &amp; EFPIA (grand no. 115749, CANCER-ID). Citation Format: Dan Brudzewsky, Rita Lampignano, Sabrina Weber, Alexander Sartori, Sumitra Mohan, Yves Konigshofer, Lorn Davis, Dominic Rothwell, Ed Schuuring, Russell K. Garlick, Thomas Schlange, Ellen Heitzer. Multicenter evaluation of circulating tumor DNA assays [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 438.

On-farm wastewater treatment using biochar from local agroresidues reduces pathogens from irrigation water for safer food production in developing countries

In this study, the suitability of an anaerobic biofilter (AnBF) as an efficient and low-cost wastewater treatment for safer irrigation water production for Sub-Saharan Africa was investigated. To determine the influence of different ubiquitous available materials on the treatment efficiency of the AnBF, rice husks and their pyrolysed equivalent, rice husk biochar, were used as filtration media and compared with sand as a common reference material. Raw sewage from a municipal full-scale wastewater treatment plant pretreated with an anaerobic filter (AF) was used in this experiment. The filters were operated at 22 °C room temperature with a hydraulic loading rate of 0.05 m·h−1 for 400 days. The mean organic loading rate (OLR) of the AF was 194 ± 74 and 63 ± 16 gCOD·m−3·d−1 for the AnBF. Fecal indicator bacteria (FIB) (up to 3.9 log10-units), bacteriophages (up to 2.7 log10-units), chemical oxygen demand (COD) (up to 94%) and turbidity (up to 97%) could be significantly reduced. Additionally, the essential plant nutrients nitrogen and phosphorous were not significantly affected by the water treatment. Overall, the performance of the biochar filters was significantly better than or equal to the sand and rice husk filters. By using the treated wastewater for irrigating lettuce plants in a pot experiment, the contamination with FIB was >2.5 log-units lower (for most of the plants below the detection limit of 5.6 MPN per gram fresh weight) than for plants irrigated with raw wastewater. Respective soil samples were minimally contaminated and nearly in the same range as that of tap water.

Shear-induced platelet adherence and activation in an in-vitro dynamic multiwell-plate system.

Circulating blood cells are prone to varying flow conditions when contacting cardiovascular devices. For a profound understanding of the complex interplay between the blood components/cells and cardiovascular implant surfaces, testing under varying shear conditions is required. Here, we study the influence of arterial and venous shear conditions on the in vitro evaluation of the thrombogenicity of polymer-based implant materials.Medical grade poly(dimethyl siloxane) (PDMS), polyethylene terephthalate (PET) and polytetrafluoroethylene (PTFE) films were included as reference materials. The polymers were exposed to whole blood from healthy humans. Blood was agitated orbitally at low (venous shear stress: 2.8 dyne · cm-2) and high (arterial shear stress: 22.2 dyne · cm-2) agitation speeds in a well-plate based test system. Numbers of non-adherent platelets, platelet activation (P-Selectin positive platelets), platelet function (PFA100 closure times) and platelet adhesion (laser scanning microscopy (LSM)) were determined.Microscopic data and counting of the circulating cells revealed increasing numbers of material-surface adherent platelets with increasing agitation speed. Also, activation of the platelets was substantially increased when tested under the high shear conditions (P-Selectin levels, PFA-100 closure times). At low agitation speed, the platelet densities did not differ between the three materials. Tested at the high agitation speed, lowest platelet densities were observed on PDMS, intermediate levels on PET and highest on PTFE. While activation of the circulating platelets was affected by the implant surfaces in a similar manner, PFA closure times did not reflect this trend.Differences in the thrombogenicity of the studied polymers were more pronounced when tested at high agitation speed due to the induced shear stresses. Testing under varying shear stresses, thus, led to a different evaluation of the implant thrombogenicity, which emphasizes the need for testing under various flow conditions. Our data further confirmed earlier findings where the same reference implants were tested under static (and not dynamic) conditions and with fresh human platelet rich plasma instead of whole blood. This supports that the application of common reference materials may improve inter-study comparisons, even under varying test conditions.

Calibration Using a Single-Point External Reference Material Harmonizes Quantitative Mass Spectrometry Proteomics Data between Platforms and Laboratories.

Mass spectrometry (MS) measurements are not inherently calibrated. Researchers use various calibration methods to assign meaning to arbitrary signal intensities and improve precision. Internal calibration (IC) methods use internal standards (IS) such as synthesized or recombinant proteins or peptides to calibrate MS measurements by comparing endogenous analyte signal to the signal from known IS concentrations spiked into the same sample. However, recent work suggests that using IS as IC introduces quantitative biases that affect comparison across studies because of the inability of IS to capture all sources of variation present throughout an MS workflow. Here, we describe a single-point external calibration strategy to calibrate signal intensity measurements to a common reference material, placing MS measurements on the same scale and harmonizing signal intensities between instruments, acquisition methods, and sites. We demonstrate data harmonization between laboratories and methodologies using this generalizable approach.

Efficient Low-Cost Anaerobic Treatment of Wastewater Using Biochar and Woodchip Filters

Access to improved sanitation is often lacking in many low-income countries, and approximately 90% of the sewage is discharged without treatment into receiving water bodies. The aim of this study was the development and evaluation of an efficient low-cost wastewater treatment system for developing countries. Biochar and woodchips, potential locally available and inexpensive materials, were used for anaerobic wastewater filtration and their suitability evaluated in comparison to gravel as a common reference material. Filters were fed with raw sewage from a municipal full-scale wastewater treatment plant in Germany at 22 °C room temperature with a hydraulic loading rate (HLR) of 0.05 m∙h−1. This resulted in a mean organic loading rate (OLR) of 252 gCOD∙m−3∙d−1 and a mean organic surface load of 456 gCOD∙m−2∙d−1. To determine the influence of different filter materials, the removal efficiency of chemical oxygen demand (COD), total organic carbon (TOC), turbidity, and faecal indicator bacteria (FIB) E. coli and enterococci were tested. It was found that COD (up to 90%), TOC (up to 80%), FIB (up to 1.7 log10-units), and turbidity (effluent turbidity below 35 NTU) could be significantly reduced. The findings of this study demonstrate the potential of anaerobic filters (AFs) for wastewater treatment in low-income countries to reduce water pollution and comprehensively improve water quality. The performance of biochar filters was significantly better over the entire experiment compared to woodchip and gravel filters with respect to COD, TOC, turbidity, and FIB removal, indicating the superior properties of biochar for wastewater treatment.