Next Generation Sequencing in Newborn Screening in the United Kingdom National Health Service.

Next generation DNA sequencing (NGS) has the potential to improve the diagnostic and prognostic utility of newborn screening programmes. This study assesses the feasibility of automating NGS on dried blood spot (DBS) DNA in a United Kingdom National Health Service (UK NHS) laboratory. An NGS panel targeting the entire coding sequence of five genes relevant to disorders currently screened for in newborns in the UK was validated on DBS DNA. An automated process for DNA extraction, NGS and bioinformatics analysis was developed. The process was tested on DBS to determine feasibility, turnaround time and cost. The analytical sensitivity of the assay was 100% and analytical specificity was 99.96%, with a mean 99.5% concordance of variant calls between DBS and venous blood samples in regions with ≥30× coverage (96.8% across all regions; all variant calls were single nucleotide variants (SNVs), with indel performance not assessed). The pipeline enabled processing of up to 1000 samples a week with a turnaround time of four days from receipt of sample to reporting. This study concluded that it is feasible to automate targeted NGS on routine DBS samples in a UK NHS laboratory setting, but it may not currently be cost effective as a first line test.

Soy protein recovery in a solvent-free process using continuous liquid-solid circulating fluidized bed ion exchanger.

Soy protein concentrates and soy protein isolates act as ingredients in bakery, meat and dairy products, baby formulas, starting materials for spun textured vegetable products, and other nutritional supplements. In this study, the effectiveness of a liquid-solid circulating fluidized bed (LSCFB) ion exchanger is demonstrated for the recovery of soluble soy proteins from full fat and defatted soy flour. Under steady-state operating conditions, about 50% of the proteins could be recovered from the feed streams entering the ion exchanger. The LSCFB was shown to be a promising system for the recovery of soy protein from both defatted and full fat soy flour solutions. As the ion exchange process captures dissolved proteins, the system may offer a less damaging form of processing compared with the acid precipitation process where soy protein aggregates form and functionality is affected. In addition, the LSCFB allows simultaneous adsorption and desorption of the proteins allowing for a continuous operation. No prefiltration of feed containing suspended particles is required as well, because fluidization is used in place of packed bed technology to improve on current ion exchange processes.

A template search reveals mechanistic similarities and differences in beta-ketoacyl synthases (KAS) and related enzymes.

A detailed comparison of the active sites in beta-ketoacyl synthases (KAS) and related enzymes has been made. Using three-dimensional templates of the three catalytic residues to scan the protein structural database reveals differences in both the geometry and the catalytic role of equivalent residues in different members of the family. The template based on the catalytic cysteine and two histidines in the KAS I and II is totally specific for this family, with no false hits. However, the role of the histidines in catalysis is different between KAS I/II and thiolase on the one hand and KAS III/chalcone synthase on the other. In contrast, a template comprising only cysteine and one histidine is not specific with many hits including members of the KAS family, metal binding sites, other active sites in nonhomologous proteins, and some "random" nonactive sites.