Comparing product category rules from different programs: learned outcomes towards global alignment

Purpose

Product category rules (PCRs) provide category-specific guidance for estimating and reporting product life cycle environmental impacts, typically in the form of environmental product declarations and product carbon footprints. Lack of global harmonization between PCRs or sector guidance documents has led to the development of duplicate PCRs for the same products. Differences in the general requirements (e.g., product category definition, reporting format) and LCA methodology (e.g., system boundaries, inventory analysis, allocation rules, etc.) diminish the comparability of product claims.

Methods

A comparison template was developed to compare PCRs from different global program operators. The goal was to identify the differences between duplicate PCRs from a broad selection of product categories and propose a path toward alignment. We looked at five different product categories: milk/dairy (two PCRs), horticultural products (three PCRs), wood?Cparticleboard (two PCRs), and laundry detergents (four PCRs).

Results and discussion

Disparity between PCRs ranged from broad differences in scope, system boundaries, and impacts addressed (e.g., multi-impact vs. carbon footprint only) to specific differences of technical elements. The differences primarily reflected the different purposes of the PCR (e.g., label/report), the different standards they were based on (e.g., ISO 14025/PAS 2050), the use of different product categorization systems, or simply the result of being developed independently. Differing degrees of specificity and terminology between PCRs allowed for varied interpretation??at times making direct comparison difficult. For many of the differences between PCRs, however, there was no clear rationale why they could not be consistent in the future.

Conclusions

These results were used to outline a general guidance document for global alignment of PCRs which recommends (1) alignment of PCRs for different purposes, (2) provision of guidance for the adoption of aspects of other PCRs, and (3) provision of greater specificity on content. The overall recommendations also suggest collaboration among program operators to facilitate alignment on issues that evolve from independent development.     

An institutional analysis of EPD programs and a global PCR registry

Purpose

A Product Category Rules (PCR) document specifies the quantification method and communication format of environmental impacts of a product category. To ensure neutrality and credibility of quantitative environmental information, the development of PCR documents is defined in ISO 14025. Hence, the rules are preconditions for comparative considerations and modular application of information entities and Environmental Product Declarations (EPD). However, with the growing number of EPD programs, the producers, purchasers, and consumers feel increasingly alienated in relation to the validity and legitimacy of the environmental information presented by EPDs. This results in a need for enhanced transparency of PCR development and EPD program compatibility. This article offers navigational assistance in this respect.

Methods

To identify harmonization potential, we compare PCR development regarding two aspects: quantitatively by mapping existing PCR and EPD documents, and qualitatively by comparing existing institutional structures with normative guidance. Information was gathered through an internet search and through direct correspondence with program operators.

Results and discussion

We identified 27 programs, 556 PCR documents, and 3614 EPD declarations (May 2013). There were significant differences in activity level between programs and sectors. Furthermore, the institutional structures differ widely from each other and from normative guidance on PCR development.

Conclusions

This first global PCR register guides practitioners in the search for PCR documents. The analysis of program institutional structures for PCR development and EPD verification indicates the involvement of different stakeholders on Type III environmental declarations. Regarding PCR compatibility and newly released guidance documents from the European Commission and the PCR Guidance Development Initiative, we recommend that operators (1) settle on a common (sector) categorization system, (2) implement a Stakeholder Identification Worksheet, (3) consider the mandatory involvement of consumer and environmental interests in the PCR review panel, (4) require PCR reviewers to declare potential conflicts of interest, and (5) consider installing mandatory third party verification of declarations for any external use.     

Global guidance principles for life cycle assessment databases: development of training material and other implementation activities on the publication

Purpose

The paper introduces the publication on “Global Guidance Principles for Life Cycle Assessment Databases”; it focuses on the development of training material and other implementation activities on the publication.

Methods

The document is the output of the “Shonan Guidance Principles” workshop. The publication provides guidance principles for life cycle assessment (LCA) databases; this includes how to collect raw data, how to develop datasets, and how to manage databases. The publication also addresses questions concerning data documentation and review, coordination among databases, capacity building, and future scenarios. As a next step, the publication is used to prepare training material and other implementation activities.

Results

The publication was launched at the LCM 2011 Conference. Since then outreach activities have been organized in particular in emerging economies. Further developments with regard to the guidance principles are foreseen as part of a flagship project within phase 3 of the Life Cycle Initiative. Training material is being developed that will include how to set up databases and develop datasets. The topic has been taken up by United Nations Environment Programme (UNEP) in its Rio?+?20 Voluntary Commitments: UNEP and Society of Environmental Toxicology and Chemistry (SETAC) through the UNEP/SETAC Life Cycle Initiative commit to facilitate improved access to good quality life cycle data and databases as well as expanded use of key environmental indicators that allows the measurement and monitoring of progress towards the environmental sustainability of selected product chains.

Conclusions

The adoption of the “Global Guidance Principles” publication as a de facto global standard is expected to facilitate the work of database teams, especially, in developing countries, and the collaboration in regional networks. These efforts are supported by the development of training material and other implementation activities.     

Pellicle associated adherence film above incubation broth surface - an inexpensive adjunct to recognizing Candida krusei in the laboratory

Background

Targeting Induced Local Lesions in Genomes (TILLING) is a high throughput reverse genetics tool which detects mismatches (single point mutations or small indels) in large number of individuals of mutagenized populations. Currently, TILLING is intensively used for genomics assisted molecular breeding of several crop plants for desired traits. Most commonly used platform for mutation detection is Li-COR DNA Analyzer, where PCR amplified products treated with single strand mismatch specific nuclease are resolved on denaturing gels. The molecular size of any cut product can be easily estimated by comparing with IR dye labeled markers of known sizes. Similar fluorescent dye labeled size markers are also used for several genotyping experiments. Currently, commercially available size standards are expensive and are restricted up to only 700 bp which renders estimation of products of sizes greater than 700 bases inaccurate.

Findings

A simple protocol was developed for labeling 5' end of multiple DNA size markers with fluorescent dyes. This method involves cloning a pool of different size markers of DNA in a plasmid vector. PCR amplification of plasmid using IR dye labeled universal primers generates 5' fluorescent labeled products of various sizes. The size of products constituting the ladder can be customized as per the need. The generated size markers can be used without any further purification and were found to be stable up to one year at -20°C.

Conclusions

A simple method was developed for generating fluorescent dye labeled size standards. This method can be customized to generate different size standards as per experimental needs. The protocol described can also be adapted for developing labeled size standards for detection on platforms other than Li-COR i.e. other than infra red range of the spectrum.     

Comparison of product carbon footprint standards with a case study on poinsettia (Euphorbia pulcherrima)

Purpose

A method to quantify the climate impact of products called product carbon footprint (PCF) has been gaining popularity in recent years. However, variations of this method have resulted in several competing standards to guide the carbon calculation process. The aim of the current paper was to compare PCF results when calculated according to the different standards.

Methods

The three leading PCF standards are Publicly Available Specification (PAS) 2050:2011, ISO.DIN 2 14067 and Product Life Cycle Accounting and Reporting Standard (PARS) 2011. These standards were compared conceptually, and a case study was performed in which the PCF of a poinsettia plant produced in Germany was calculated according to all three standards.

Results and discussion

The PCF results were 0.45–0.50, 0.53–0.58 and 0.53–0.59 kg carbon dioxide equivalent according to PAS 2050:2011, ISO.DIN 2 14067 and PARS 2011, respectively. According to all standards, the life cycle stage contributing the most greenhouse gases (GHGs) was the production of the poinsettia plant, and the single process with the highest emissions was the electricity use in the production. It was found that if nonrenewable fuels were used for heating instead of wood chips, then heating would be the highest GHG contributor—accounting for over 80 % of emissions of the total PCF.

Conclusions

A key finding was that both the production system used and the decisions taken by the person carrying out the PCF calculation result in greater differences in the PCF result than the use of different standards. Differences among the three standards could be harmonised by more specific cut-off rules and exclusion criteria with the publication of ISO.DIN 2 14067, as well as the development and use of product category rules.     

Life Cycle Impact Assessment—where we are, trends, and next steps: a late report from a UNEP/SETAC Life Cycle Initiative workshop and a few updates from recent developments

Purpose

The paper provides a late report from the United Nations Environment Program (UNEP)/Society of Environmental Toxicology and Chemistry (SETAC) Life Cycle Initiative workshop “Life Cycle Impact Assessment (LCIA)—where we are, trends, and next steps;” it embeds this report into recent development with regard to the envisaged development of global guidance on environmental life cycle impact assessment indicators and related methodologies.

Methods

The document is the output of the UNEP/SETAC Life Cycle Initiative’s workshop on “Life Cycle Impact Assessment—where we are, trends, and next steps.” The presentations and discussions held during the workshop reviewed the first two phases of the Life Cycle Initiative and provided an overview of current LCIA activities being conducted by the Initiative, governments and academia, as well as corporate approaches. The outcomes of the workshop are reflected in light of the implementation of the strategy for Phase 3 of the Life Cycle Initiative.

Results

The range of views provided during the workshop indicated different user needs, with regards to, amongst other things, the required complexity of the LCIA methodology, associated costs, and the selection of LCIA categories depending on environmental priorities. The workshop’s results signified a number of potential focus areas for Phase 3 of the Initiative, including capacity building efforts concerning LCIA in developing countries and emerging economies, the preparation of training materials on LCIA, the production of global guidance on LCIA, and the potential development of a broader sustainability indicators framework.

Conclusions

These suggestions have been taken into account in the strategy for Phase 3 of the Life Cycle Initiative in two flagship projects, one on global capability development on life cycle approaches and the other on global guidance on environmental life cycle impact assessment indicators. In the context of the latter project, first activities are being organized and planned. Moreover, UNEP has included the recommendations in its Rio + 20 Voluntary Commitments: UNEP and SETAC through the UNEP/SETAC Life Cycle Initiative commit to facilitate improved access to good quality life cycle data and databases as well as expanded use of key environmental indicators that allows the measurement and monitoring of progress towards the environmental sustainability of selected product chains.     

Mesoporous silicon nanotechnology for cancer applications

Abstract

The commonest genetic defects leading to α-thalassemia are genomic deletions encompassing one or both α-globin loci. The aim of this work was to evaluate the interest of melting curves characteristics obtained in gap real-time polymerase chain reaction for the diagnosis of α-thalassemia deletions. Five patients with known deletional α-thalassemia, and five control subjects were tested. The melting temperature and melting curve shapes of the PCR products allowed us to discriminate controls from α-thalassemic patients. This promising strategy should be further tested with samples carrying other α-thalassemic deletions.

Background

The commonest genetic defects leading to α-thalassemia are genomic deletions encompassing one or both α-globin loci. The aim of this work was to evaluate the interest of melting curves characteristics obtained in gap real-time polymerase chain reaction for the diagnosis of α-thalassemia deletions.

Methods

Five patients with known deletional α thalassemia, —(MED)(2), —(SEA)(2), —(FIL)(1), and five control subjects were tested. Specific primers were used for each deletion. Real-time-gap PCRs, using SYBER-Green mix, were performed on a « light-cycler 480 from Roche® ». At the melting temperature, the released of the SYBER-Green dye bonded to double strained DNA is associated with a dramatic decrease in fluorescence intensity. The rate of this variation was determined by plotting the negative derivative of fluorescence vs temperature (lightcycler 480 software, Roche®).

Results

The melting temperature (Tm) and melting curve shapes of the PCR products derived from α-thalassemic patients or control subjects, allowed us to discriminate controls from α-thalassemic patients (eg —Phil Tm = 91°C whereas control Tm = 81 and 85,6°C). Some non specific amplified products are still present, probably because of the high GC content and high homology of sequences in the α-globin cluster.

Conclusion

This promising strategy should be further tested with samples carrying other α-thalassemic deletions. Nevertheless it appears reliable, cost-effective and safe offering additional benefits including minimal labor, rapid turnaround time and decreased risk of PCR carryover contamination. It may be one alternative technology available for routine diagnosis of all types of deletional α-thalassemia.     

Selective measurement of α smooth muscle actin: why β-actin can not be used as a housekeeping gene when tissue fibrosis occurs

Background

Prevalence of fibroproliferative diseases, including chronic kidney disease is rapidly increasing and has become a major public health problem worldwide. Fibroproliferative diseases are characterized by increased expression of α smooth muscle actin (α-SMA) that belongs to the family of the six conserved actin isoforms showing high degree homology. The aim of the present study was to develop real-time PCRs that clearly discriminate α-SMA and ß-actin from other actin isoforms.

Results

Real-time PCRs using self-designed mouse, human and rat specific α-SMA or ß-actin primer pairs resulted in the specific amplification of the artificial DNA templates corresponding to mouse, human or rat α-SMA or ß-actin, however ß-actin showed cross-reaction with the housekeeping γ-cyto-actin. We have shown that the use of improperly designed literary primer pairs significantly affects the results of PCRs measuring mRNA expression of α-SMA or ß-actin in the kidney of mice underwent UUO.

Conclusion

We developed a set of carefully designed primer pairs and PCR conditions to selectively determine the expression of mouse, human or rat α-SMA and ß-actin isoforms. We demonstrated the importance of primer specificity in experiments where the results are normalized to the expression of ß-actin especially when fibrosis and thus increased expression of α-SMA is occur.

     

Large-scale preparation of active caspase-3 in E. coli by designing its thrombin-activatable precursors

Background

Combination of CHD (chromo-helicase-DNA binding protein)-specific polymerase chain reaction (PCR) with electrophoresis (PCR/electrophoresis) is the most common avian molecular sexing technique but it is lab-intensive and gel-required. Gender determination often fails when the difference in length between the PCR products of CHD-Z and CHD-W genes is too short to be resolved.

Results

Here, we are the first to introduce a PCR-melting curve analysis (PCR/MCA) to identify the gender of birds by genomic DNA, which is gel-free, quick, and inexpensive. Spilornis cheela hoya (S. c. hoya) and Pycnonotus sinensis (P. sinensis) were used to illustrate this novel molecular sexing technique. The difference in the length of CHD genes in S. c. hoya and P. sinensis is 13-, and 52-bp, respectively. Using Griffiths' P2/P8 primers, molecular sexing failed both in PCR/electrophoresis of S. c. hoya and in PCR/MCA of S. c. hoya and P. sinensis. In contrast, we redesigned sex-specific primers to yield 185- and 112-bp PCR products for the CHD-Z and CHD-W genes of S. c. hoya, respectively, using PCR/MCA. Using this specific primer set, at least 13 samples of S. c. hoya were examined simultaneously and the Tm peaks of CHD-Z and CHD-W PCR products were distinguished.

Conclusion

In this study, we introduced a high-throughput avian molecular sexing technique and successfully applied it to two species. This new method holds a great potential for use in high throughput sexing of other avian species, as well.     

A proof of the DBRF-MEGN method, an algorithm for deducing minimum equivalent gene networks

Background

We previously developed the DBRF-MEGN (difference-based regulation finding-minimum equivalent gene network) method, which deduces the most parsimonious signed directed graphs (SDGs) consistent with expression profiles of single-gene deletion mutants. However, until the present study, we have not presented the details of the method's algorithm or a proof of the algorithm.

Results

We describe in detail the algorithm of the DBRF-MEGN method and prove that the algorithm deduces all of the exact solutions of the most parsimonious SDGs consistent with expression profiles of gene deletion mutants.

Conclusions

The DBRF-MEGN method provides all of the exact solutions of the most parsimonious SDGs consistent with expression profiles of gene deletion mutants.     

Real-time PCR TaqMan assay for rapid screening of bloodstream infection

Background

Sepsis is one of the main causes of mortality and morbidity. The rapid detection of pathogens in blood of septic patients is essential for adequate antimicrobial therapy and better prognosis. This study aimed to accelerate the detection and discrimination of Gram-positive (GP) and Gram-negative (GN) bacteria and Candida species in blood culture samples by molecular methods.

Methods

The Real-GP®, -GN®, and -CAN® real-time PCR kit (M&D, Wonju, Republic of Korea) assays use the TaqMan probes for detecting pan-GP, pan-GN, and pan-Candida species, respectively. The diagnostic performances of the real-time PCR kits were evaluated with 115 clinical isolates, 256 positive and 200 negative blood culture bottle samples, and the data were compared to results obtained from conventional blood culture.

Results

Eighty-seven reference strains and 115 clinical isolates were correctly identified with specific probes corresponding to GP-bacteria, GN-bacteria and Candida, respectively. The overall sensitivity and specificity of the real-time PCR kit with blood culture samples were 99.6% and 89.5%, respectively.

Conclusions

The Real-GP®, -GN®, and -CAN® real-time PCR kits could be useful tools for the rapid and accurate screening of bloodstream infections (BSIs).     

Primary lung tumour visualised by transthoracic echocardiography

Background

Stroke is the third most common cause of death in the UK and the largest single cause of severe disability. Each year more than 110,000 people in England suffer from a stroke which costs the National Health Service (NHS) over GBP2.8 billion. Thus, it is imperative that patients at risk be screened for underlying carotid artery atherosclerosis.

Aim

To assess the role of carotid ultrasound in different carotid screening programmes.

Methods

A literature overview was carried out by using PubMed search engine, to identify different carotid screening programmes that had used ultrasound scan as a screening tool.

Results

It appears that the carotid ultrasound is an effective method for screening carotid artery disease in community as it effectively predicts the presence of stenosis with high accuracy. There is a need for primary care to recommend high risk patients for regular screening, to reduce stroke and transient ischemic attack (TIA) related morbidity and mortality.

Conclusion

Screening programmes using carotid ultrasonography contribute to public health awareness and promotion which in long term could potentially benefit in disease prevention and essentially promote better standards of healthcare.     

Simulation and analysis for sustainable product development

Purpose

Simulation plays a critical role in the design of products, materials, and manufacturing processes. However, there are gaps in the simulation tools used by industry to provide reliable results from which effective decisions can be made about environmental impacts at different stages of product life cycle. A holistic and systems approach to predicting impacts via sustainable manufacturing planning and simulation (SMPS) is presented in an effort to incorporate sustainability aspects across a product life cycle.

Methods

Increasingly, simulation is replacing physical tests to ensure product reliability and quality, thereby facilitating steady reductions in design and manufacturing cycles. For SMPS, we propose to extend an earlier framework developed in the Systems Integration for Manufacturing Applications (SIMA) program at the National Institute of Standards and Technology. SMPS framework has four phases, viz. design product, engineer manufacturing, engineer production system, and produce products. Each phase has its inputs, outputs, phase level activities, and sustainability-related data, metrics and tools.

Results and discussion

An automotive manufacturing scenario that highlights the potential of utilizing SMPS framework to facilitate decision making across different phases of product life cycle is presented. Various research opportunities are discussed for the SMPS framework and corresponding information models.

Conclusions

The SMPS framework built on the SIMA model has potential in aiding sustainable product development.