The use of soybean peroxidase in amperometric biosensors

Traditionally Horseradish peroxidase (HRP) the archetypal class III plant peroxidase has been investigated with respect to chemical modifications, organic solvent tolerance studies, reaction kinetics, kinetics in organic solvents, immunohistochemical and in biosensor fabrication. However HRP has many factors that prevent its use in industrial, and research areas. This is because of its inherent structure, and activity restrictions in regard to temperature, its pH range of activity, and solvents it is catalytically active in. There is another class III plant peroxidase, which has greater stability with regards to the above factors, this peroxidase is Soybean peroxidase (SBP). This is the peroxidase that is investigated in this work. Chemical modifications historically have targeted amino acid residues in the tertiary folded active form of an enzyme. HRP and many other enzymes have been chemically modified by modifying the E - N H 3 group of lysine (HRP has 3 lysines available for modification out of 5 lysines). SBP cannot be chemically modified in this fashion, because it has no lysines available for modification, out of 3 lysines). Therefore in order to attempt to successfully chemically modify SBP another approach needs to be studied. SBP is a glycosylated enzyme. By chemically modifying this catalytically inactive region of SBP, an increase in stability and activity can be achieved. Three methods of chemically modifying the carbohydrate shell, these were modifying by dextran dialdehyde, adipic acid dihydrazide (AADH), and ferrocene carboxcylic acid (FeCOOH). The dextran dialdehyde modification yielded a 2-fold modification. The AADH modification yielded a three-fold modification, but problems arose with the inhibition of the enzymes active site and the cross-linking agent. The FeCOOH modification was successful adding at least 2 extra irons to the enzyme. The modified was used in conjunction with a sensor. This modification also reduces the electron transfer distance between the enzyme and the transducer. This sensor was characterised and compared to a native SBP biosensor. Also investigated in this work was the non-heme peroxidase vanadium bromoperoxidase (VBrPO). VBrPO is a sourced enzyme, utilising vanadium at its active site. The native enzyme was characterised and the dextran dialdehyde modification was carried out on it. The modification was then characterised and compared to the native form of the enzyme

Horseradish and Soybean Peroxidases: Comparable Tools for Alternative Niches?

Horseradish and soybean peroxidases (HRP and SBP, respectively) are useful biotechnological tools. HRP is often termed the classical plant heme peroxidase, and although it has been studied for decades our understanding has deepened since its cloning and subsequent expression, which has enabled numerous mutational and protein engineering studies. SBP, however, has been neglected until recently; despite offering a real alternative to HRP that actually outperforms it in terms of stability. SBP is now used in numerous biotechnological applications, including biosensors. Review of both is timely. This article summarises and discusses the main insights into the structure and mechanism of HRP, with special emphasis on HRP mutagenesis, and outlines its use in a variety of applications. It also reviews current knowledge and applications to date of SBP, particularly biosensors. The final paragraphs speculate on the future of plant heme-based peroxidases, with probable trends outlined and explored

The TURis system for transurethral resection of the prostate: a NICE medical technology guidance

The transurethral resection in saline (TURis) system was notified by the company Olympus Medical to the National Institute of Health and Care Excellence’s (NICE’s) Medical Technologies Evaluation Programme. Following selection for medical technologies guidance, the company developed a submission of clinical and economic evidence for evaluation. TURis is a bipolar surgical system for treating men with lower urinary tract symptoms due to benign prostatic enlargement. The comparator is any monopolar transurethral resection of the prostate (mTURP) system. Cedar, a collaboration between Cardiff and Vale University Health Board, Cardiff University and Swansea University in the UK, acted as an External Assessment Centre (EAC) for NICE to independently critique the company’s submission of evidence. Eight randomised trials provided evidence for TURis, demonstrating efficacy equivalent to that of mTURP for improvement of symptoms. The company presented meta-analyses of key outcome measures, and the EAC made methodological modifications in response to the heterogeneity of the trial data. The EAC analysis found that TURis substantially reduced the relative risks of transurethral resection syndrome (relative risk 0.18 [95 % confidence interval 0.05–0.62]) and blood transfusion (relative risk 0.35 [95 % confidence interval 0.19–0.65]). The company provided a de novo economic model comparing TURis with mTURP. The EAC critiqued the model methodology and made modifications. This found TURis to be cost saving at £70.55 per case for existing Olympus customers and cost incurring at £19.80 per case for non-Olympus customers. When an additional scenario based on the only available data on readmission (due to any cause) from a single trial was modelled, the estimated cost saving per case was £375.02 for existing users of Olympus electrosurgery equipment and £284.66 per case when new Olympus equipment would need to be purchased. Meta-analysis of eight randomised trials showed that TURis is associated with a statistically significantly reduced risk of transurethral resection syndrome and a reduced need for blood transfusion—two factors that may drive cost saving for the National Health Service. The clinical data are equivocal as to whether TURis shortens the hospital stay. Limited data from a single study suggest that TURis may reduce the rate of readmission after surgery. The NICE guidance supports adoption of the TURis technology for performing transurethral resection of the prostate in men with lower urinary tract symptoms due to benign prostatic enlargemen

Horseradish and soybean peroxidases: comparable tools for alternative niches?

Horseradish and soybean peroxidases (HRP and SBP, respectively) are useful biotechnological tools. HRP is often termed the classical plant heme peroxidase and although it has been studied for decades, our understanding has deepened since its cloning and subsequent expression, enabling numerous mutational and protein engineering studies. SBP, however, has been neglected until recently, despite offering a real alternative to HRP: SBP actually outperforms HRP in terms of stability and is now used in numerous biotechnological applications, including biosensors. Review of both is timely. This article summarizes and discusses the main insights into the structure and mechanism of HRP, with special emphasis on HRP mutagenesis, and outlines its use in a variety of applications. It also reviews the current knowledge and applications to date of SBP, particularly biosensors. The final paragraphs speculate on the future of plant heme-based peroxidases, with probable trends outlined and explored

The TURis System for Transurethral Resection of the Prostate: A NICE Medical Technology Guidance

The transurethral resection in saline (TURis) system was notified by the company Olympus Medical to the National Institute of Health and Care Excellence’s (NICE’s) Medical Technologies Evaluation Programme. Following selection for medical technologies guidance, the company developed a submission of clinical and economic evidence for evaluation. TURis is a bipolar surgical system for treating men with lower urinary tract symptoms due to benign prostatic enlargement. The comparator is any monopolar transurethral resection of the prostate (mTURP) system. Cedar, a collaboration between Cardiff and Vale University Health Board, Cardiff University and Swansea University in the UK, acted as an External Assessment Centre (EAC) for NICE to independently critique the company’s submission of evidence. Eight randomised trials provided evidence for TURis, demonstrating efficacy equivalent to that of mTURP for improvement of symptoms. The company presented meta-analyses of key outcome measures, and the EAC made methodological modifications in response to the heterogeneity of the trial data. The EAC analysis found that TURis substantially reduced the relative risks of transurethral resection syndrome (relative risk 0.18 [95 % confidence interval 0.05–0.62]) and blood transfusion (relative risk 0.35 [95 % confidence interval 0.19–0.65]). The company provided a de novo economic model comparing TURis with mTURP. The EAC critiqued the model methodology and made modifications. This found TURis to be cost saving at £70.55 per case for existing Olympus customers and cost incurring at £19.80 per case for non-Olympus customers. When an additional scenario based on the only available data on readmission (due to any cause) from a single trial was modelled, the estimated cost saving per case was £375.02 for existing users of Olympus electrosurgery equipment and £284.66 per case when new Olympus equipment would need to be purchased. Meta-analysis of eight randomised trials showed that TURis is associated with a statistically significantly reduced risk of transurethral resection syndrome and a reduced need for blood transfusion—two factors that may drive cost saving for the National Health Service. The clinical data are equivocal as to whether TURis shortens the hospital stay. Limited data from a single study suggest that TURis may reduce the rate of readmission after surgery. The NICE guidance supports adoption of the TURis technology for performing transurethral resection of the prostate in men with lower urinary tract symptoms due to benign prostatic enlargemen

Biologic Phenotyping of the Human Small Airway Epithelial Response to Cigarette Smoking

BACKGROUND: The first changes associated with smoking are in the small airway epithelium (SAE). Given that smoking alters SAE gene expression, but only a fraction of smokers develop chronic obstructive pulmonary disease (COPD), we hypothesized that assessment of SAE genome-wide gene expression would permit biologic phenotyping of the smoking response, and that a subset of healthy smokers would have a "COPD-like" SAE transcriptome. METHODOLOGY/PRINCIPAL FINDINGS: SAE (10th-12th generation) was obtained via bronchoscopy of healthy nonsmokers, healthy smokers and COPD smokers and microarray analysis was used to identify differentially expressed genes. Individual responsiveness to smoking was quantified with an index representing the % of smoking-responsive genes abnormally expressed (I(SAE)), with healthy smokers grouped into "high" and "low" responders based on the proportion of smoking-responsive genes up- or down-regulated in each smoker. Smokers demonstrated significant variability in SAE transcriptome with I(SAE) ranging from 2.9 to 51.5%. While the SAE transcriptome of "low" responder healthy smokers differed from both "high" responders and smokers with COPD, the transcriptome of the "high" responder healthy smokers was indistinguishable from COPD smokers. CONCLUSION/SIGNIFICANCE: The SAE transcriptome can be used to classify clinically healthy smokers into subgroups with lesser and greater responses to cigarette smoking, even though these subgroups are indistinguishable by clinical criteria. This identifies a group of smokers with a "COPD-like" SAE transcriptome

CCDC 928436: Experimental Crystal Structure Determination

Related Article: A.N.Carolan,G.M.Cockrell,N.J.Williams,Gang Zhang,D.G.VanDerveer,Hee-Seung Lee,R.P.Thummel,R.D.Hancock|2013|Inorg.Chem.|52|15|doi:10.1021/ic3002509,An entry from the Cambridge Structural Database, the world’s repository for small molecule crystal structures. The entry contains experimental data from a crystal diffraction study. The deposited dataset for this entry is freely available from the CCDC and typically includes 3D coordinates, cell parameters, space group, experimental conditions and quality measures

CCDC 928437: Experimental Crystal Structure Determination

Related Article: A.N.Carolan,G.M.Cockrell,N.J.Williams,Gang Zhang,D.G.VanDerveer,Hee-Seung Lee,R.P.Thummel,R.D.Hancock|2013|Inorg.Chem.|52|15|doi:10.1021/ic3002509,An entry from the Cambridge Structural Database, the world’s repository for small molecule crystal structures. The entry contains experimental data from a crystal diffraction study. The deposited dataset for this entry is freely available from the CCDC and typically includes 3D coordinates, cell parameters, space group, experimental conditions and quality measures

Electronic Appendix 1 from Extending the durability of cultivar resistance by limiting epidemic growth rates

Detailed model structure and parameterization