Formation of urea and guanidine by irradiation of ammonium cyanide

Summary Aqueous solutions of ammonium cyanide yield urea, cyanamide and guanidine when exposed to sunlight or an unfiltered 254 muµ ultraviolet source. The prebiotic significance of these results is discussed.     

The development of Luminomaster™, a fully automated chemiluminescent enzyme immunoassay system

We have developed a fully automated discrete chemiluminescent heterogeneous enzyme immunoassay system called Luminomaster?. The characteristics of this analyser are: 120 test/h throughput, 14 test menu, wide dynamic range, automated sample dilution, automatic retest, communication with a host central processing unit (CPU) and connection with sample transfer system.     

Isolation and characterization of circulating tumor cells using a novel workflow combining the CellSearch® system and the CellCelector™

Circulating tumor cells (CTC) are rare cells which have left the primary tumor to enter the blood stream. Although only a small CTC subgroup is capable of extravasating, the presence of CTCs is associated with an increased risk of metastasis and a shorter overall survival. Understanding the heterogeneous CTC biology will optimize treatment decisions and will thereby improve patient outcome. For this, robust workflows for detection and isolation of CTCs are urgently required. Here, we present a workflow to characterize CTCs by combining the advantages of both the CellSearch^® and the CellCelector? micromanipulation system. CTCs were isolated from CellSearch^® cartridges using the CellCelector? system and were deposited into PCR tubes for subsequent molecular analysis (whole genome amplification (WGA) and massive parallel multigene sequencing). By a CellCelector? screen we reidentified 97% of CellSearch^® SKBR‐3 cells. Furthermore, we isolated 97% of CellSearch^®‐proven patient CTCs using the CellCelector? system. Therein, we found an almost perfect correlation of R² = 0.98 (Spearman's rho correlation, n = 20, p < 0.00001) between the CellSearch^® CTC count (n = 271) and the CellCelector? detected CTCs (n = 252). Isolated CTCs were analyzed by WGA and massive parallel multigene sequencing. In total, single nucleotide polymorphisms (SNPs) could be detected in 50 genes in seven CTCs, 12 MCF‐7, and 3 T47D cells, respectively. Taken together, CTC quantification via the CellCelector? system ensures a comprehensive detection of CTCs preidentified by the CellSearch^® system. Moreover, the isolation of CTCs after CellSearch^® using the CellCelector? system guarantees for CTC enrichment without any contaminants enabling subsequent high throughput genomic analyses on single cell level. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 33:125–132, 2017     

Performance of an ecological treatment system at three strengths of dairy wastewater loading

Ecological treatment systems, which rely on renewable resources, have successfully treated municipal and industrial effluents with reduced costs compared to conventional methods, but their capacity to treat dairy wastewater is unknown. In order for ecological treatment systems to be practical for agriculture they must be able to treat a significant portion of a dairy's daily wastewater production. In this study, the impact of three strengths of dairy wastewater on effluent water quality was assessed. Three ratios of wastewater and city water—(1) one part wastewater:three parts city water, (2) one part wastewater:one part city water, and (3) two parts wastewater:one part city water—were each pumped into an ecological treatment system. Influent and effluent water samples were analyzed for PO₄-P, TP, TN, NH₄-N, NO₃-N, total suspended solids (TSS), and carbonaceous biochemical oxygen demand (CBOD₅). Influent dairy wastewater volumetric loading rates were much greater than those of municipal wastewater. Regardless of influent wastewater strength, concentrations of all measured variables were significantly reduced between the influent and effluent of the ecological treatment system. At the lowest wastewater strength, PO₄-P was reduced 39%, TN 83%, and NH₄-N 89%, while at the highest wastewater strength, PO₄-P was reduced 41%, TN 79%, and NH₄-N 70%. Increased wastewater strength required greater aerobic treatment volume to reduce concentrations of NH₄-N and CBOD₅.     

Inhibition of protein carbamylation in urea solution using ammonium-containing buffers

Urea solution is one of the most commonly employed protein denaturants for protease digestion in proteomic studies. However, it has long been recognized that urea solution can cause carbamylation at the N termini of proteins/peptides and at the side chain amino groups of lysine and arginine residues. Protein/peptide carbamylation blocks protease digestion and affects protein identification and quantification in mass spectrometry analysis by blocking peptide amino groups from isotopic/isobaric labeling and changing peptide charge states, retention times, and masses. In addition, protein carbamylation during sample preparation makes it difficult to study in vivo protein carbamylation. In this study, we compared the peptide carbamylation in urea solutions of different buffers and found that ammonium-containing buffers were the most effective buffers to inhibit protein carbamylation in urea solution. The possible mechanism of carbamylation inhibition by ammonium-containing buffers is discussed, and a revised procedure for the protease digestion of proteins in urea and ammonium-containing buffers was developed to facilitate its application in proteomic research.