Chilling slows anaerobic metabolism to improve anoxia tolerance of insects

Background

Insects are renowned for their ability to survive anoxia. Anoxia tolerance may be enhanced during chilling through metabolic suppression.

Aims

Here, the metabolomic response of insects to anoxia, both with and without chilling, for different durations (12–36 h) was examined to assess the potential cross-tolerance mechanisms.

Results

Chilling during anoxia (cold anoxia) significantly improved survival relative to anoxia at warmer temperatures. Reduced intermediate metabolites and increased lactic acid, indicating a switch to anaerobic metabolism, were characteristic of larvae in anoxia.

Conclusions

Anoxia tolerance was correlated survival improvements after cold anoxia were correlated with a reduction in anaerobic metabolism.

     

Mitochondrial aconitase is a key regulator of energy production for growth and protein expression in Chinese hamster ovary cells

Introduction

Mammalian cells like Chinese hamster ovary (CHO) cells are routinely used for production of recombinant therapeutic proteins. Cells require a continuous supply of energy and nutrients to sustain high cell densities whilst expressing high titres of recombinant proteins. Cultured mammalian cells are primarily dependent on glucose and glutamine metabolism for energy production.

Objectives

The TCA cycle is the main source of energy production and its continuous flow is essential for cell survival. Modulated regulation of TCA cycle can affect ATP production and influence CHO cell productivity.

Methods

To determine the key metabolic reactions of the cycle associated with cell growth in CHO cells, we transiently silenced each gene of the TCA cycle using RNAi.

Results

Silencing of at least four TCA cycle genes was detrimental to CHO cell growth. With an exception of mitochondrial aconitase (or Aco2), all other genes were associated with ATP production reactions of the TCA cycle and their resulting substrates can be supplied by other anaplerotic and cataplerotic reactions. This study is the first of its kind to have established key role of aconitase gene in CHO cells. We further investigated the temporal effects of aconitase silencing on energy production, CHO cell metabolism, oxidative stress and recombinant protein production.

Conclusion

Transient silencing of mitochondrial aconitase inhibited cell growth, reduced ATP production, increased production of reactive oxygen species and reduced cell specific productivity of a recombinant CHO cell line by at least twofold.

     

Untargeted metabolomics suffers from incomplete raw data processing

Introduction

Untargeted metabolomics is a powerful tool for biological discoveries. To analyze the complex raw data, significant advances in computational approaches have been made, yet it is not clear how exhaustive and reliable the data analysis results are.

Objectives

Assessment of the quality of raw data processing in untargeted metabolomics.

Methods

Five published untargeted metabolomics studies, were reanalyzed.

Results

Omissions of at least 50 relevant compounds from the original results as well as examples of representative mistakes were reported for each study.

Conclusion

Incomplete raw data processing shows unexplored potential of current and legacy data.

     

Citrate usage in the leading causes of blindness: new possibilities for the old metabolite

Introduction

Citrate is an old metabolite which is best known for the role in the Krebs cycle. Citrate is widely used in many branches of medicine. In ophthalmology citrate is considered as a therapeutic agent and an useful diagnostic tool—biomarker.

Objectives

To summarize the published literature on citrate usage in the leading causes of blindness and highlight the new possibilities for this old metabolite.

Methods

We conducted a systematic search of the scientific literature about citrate usage in ophthalmology up to January 2018. The reference lists of identified articles were searched for providing in-depth information.

Results

This systematic review included 30 articles. The role of citrate in the leading causes of blindness is presented.

Conclusions

Citrate might help inhibit cataract progression, in case of questions confirm glaucoma diagnosis or improve cornea repair treatment as adjuvant agent (therapy of ulcerating cornea after alkali injury, crosslinking procedure). However, the knowledge about possible citrate usage in ophthalmology is not widely known. Promoting recent scientific knowledge about citrate usage in ophthalmology may not only benefit of medical improvement but may also limit economic costs caused by leading causes of blindness. Further studies on citrate usage in ophthalmology should continuously be the field of scientific interest.

     

Attributional and consequential LCA in the ILCD handbook

Purpose

This discussion article aims to highlight two problematic aspects in the International Reference Life Cycle Data System (ILCD) Handbook: its guidance to the choice between attributional and consequential modeling and to the choice between average and marginal data as input to the life cycle inventory (LCI) analysis.

Methods

We analyze the ILCD guidance by comparing different statements in the handbook with each other and with previous research in this area.

Results and discussion

We find that the ILCD handbook is internally inconsistent when it comes to recommendations on how to choose between attributional and consequential modeling. We also find that the handbook is inconsistent with much of previous research in this matter, and also in the recommendations on how to choose between average and marginal data in the LCI.

Conclusions

Because of the inconsistencies in the ILCD handbook, we recommend that the handbook be revised.

     

Optimization of fecal sample preparation for untargeted LC-HRMS based metabolomics

Introduction

Collecting feces is easy. It offers direct outcome to endogenous and microbial metabolites.

Objectives

In a context of lack of consensus about fecal sample preparation, especially in animal species, we developed a robust protocol allowing untargeted LC-HRMS fingerprinting.

Methods

The conditions of extraction (quantity, preparation, solvents, dilutions) were investigated in bovine feces.

Results

A rapid and simple protocol involving feces extraction with methanol (1/3, M/V) followed by centrifugation and a step filtration (10 kDa) was developed.

Conclusion

The workflow generated repeatable and informative fingerprints for robust metabolome characterization.

     

A life cycle comparison of disposal and beneficial use of coal combustion products in Florida

Background, Goal, and Scope

Currently, only 40%, or 44.5 million metric tons, of coal combustion products (CCPs) generated in the United States each year by electric utilities are diverted from disposal in landfills or surface impoundments and recycled. Despite promising economic and environmental savings, there has been scant attention devoted to assessing life cycle impacts of CCP disposal and beneficial use. The objective of this paper is to present a life cycle inventory considering two cases of CCP management, including the stages of coal mining and preparation, coal combustion, CCP disposal, and CCP beneficial use. Six beneficial uses were considered: concrete production, structural fills, soil amendments, road construction, blasting grit and roofing granules, and wallboard.

Methods

Primary data for raw material inputs and emissions of all stages considered were obtained from surveys and site visits of coal-burning utilities in Florida conducted in 2002, and secondary data were obtained from various published sources and from databases available in SimaPro 5.1 (PRé Consultants, Amersfoort, The Netherlands).

Results

Results revealed that 50 percent of all CCPs produced, or 108 kg per 1,000 kg of coal combusted, are diverted for application in a beneficial use; however, the relative amounts sold by each utility is dependent on the process operating parameters, air emission control devices, and resulting quality of CCP. Diversion of 50% of all CCPs to beneficial use applications yields a decrease in the total raw materials requirements (with the exception of gravel and iron) and most emissions to air, water, and land, as compared to 100% disposal.

Discussion

The greatest reduction of raw materials was attributed to replacing Portland cement with fly ash, using bottom ash as an aggregate in concrete production and road construction in place of natural materials, and substituting FGD gypsum for natural gypsum in wallboard. The use of fly ash as cementitious material in concrete also promised significant reductions in emissions, particularly the carbon dioxide that would be generated from Portland cement production. Beneficial uses of fly ash and gypsum showed reductions of emissions to water (particularly total dissolved solids) and emissions of metals to land, although these reductions were small compared to simply diverting 50% of all CCPs from landfills or surface impoundments.

Conclusions

This life cycle inventory (LCI) provides the foundation for assessing the impacts of CCP disposal and beneficial use. Beneficial use of CCPs is shown here to yield reductions in raw material requirements and various emissions to all environmental compartments, with potential tangible savings to human health and the environment.

Recommendations and Perspectives

Extension of this life cycle inventory to include impact assessment and sensitivity analysis will enable a determination of whether the savings in emissions reported here actually result in significant improvements in environmental and human health impacts.

     

Acute effect of cycling intervention on carotid arterial hemodynamics: basketball athletes versus sedentary controls

Objective

To compare the acute effects of a cycling intervention on carotid arterial hemodynamics between basketball athletes and sedentary controls.

Methods

Ten young long-term trained male basketball athletes (BA) and nine age-matched male sedentary controls (SC) successively underwent four bouts of exercise on a bicycle ergometer at the same workload. Hemodynamic variables at right common carotid artery were determined at rest and immediately following each bout of exercise. An ANCOVA was used to compare differences between the BA and SC groups at rest and immediately following the cycling intervention. The repeated ANOVA was used to assess differences between baseline and each bout of exercise within the BA or SC group.

Results

In both groups, carotid hemodynamic variables showed significant differences at rest and immediately after the cycling intervention. At rest, carotid arterial stiffness was significantly decreased and carotid arterial diameter was significantly increased in the BA group as compared to the SC group. Immediately following the cycling intervention, carotid arterial stiffness showed no obvious changes in the BA group but significantly increased in the SC group. It is worth noting that while arterial stiffness was lower in the BA group than in the SC group, the oscillatory shear index (OSI) was significantly higher in the BA group than in the SC group both at rest and immediately following the cycling intervention.

Conclusion

Long-term basketball exercise had a significant impact on common carotid arterial hemodynamic variables not only at rest but also after a cycling intervention. The role of OSI in the remodeling of arterial structure and function in the BA group at rest and after cycling requires clarification.

     

WRN-targeted therapy using inhibitors NSC 19630 and NSC 617145 induce apoptosis in HTLV-1-transformed adult T-cell leukemia cells

Background

Human T-cell leukemia virus type 1 (HTLV-1) infection is associated with adult T-cell leukemia/lymphoma (ATLL), a lymphoproliferative malignancy with a dismal prognosis and limited therapeutic options. Recent evidence shows that HTLV-1-transformed cells present defects in both DNA replication and DNA repair, suggesting that these cells might be particularly sensitive to treatment with a small helicase inhibitor. Because the “Werner syndrome ATP-dependent helicase” encoded by the WRN gene plays important roles in both cellular proliferation and DNA repair, we hypothesized that inhibition of WRN activity could be used as a new strategy to target ATLL cells.

Methods

Our analysis demonstrates an apoptotic effect induced by the WRN helicase inhibitor in HTLV-1-transformed cells in vitro and ATL-derived cell lines. Inhibition of cellular proliferation and induction of apoptosis were demonstrated with cell cycle analysis, XTT proliferation assay, clonogenic assay, annexin V staining, and measurement of mitochondrial transmembrane potential.

Results

Targeted inhibition of the WRN helicase induced cell cycle arrest and apoptosis in HTLV-1-transformed leukemia cells. Treatment with NSC 19630 (WRN inhibitor) induces S-phase cell cycle arrest, disruption of the mitochondrial membrane potential, and decreased expression of anti-apoptotic factor Bcl-2. These events were associated with activation of caspase-3-dependent apoptosis in ATL cells. We identified some ATL cells, ATL-55T and LMY1, less sensitive to NSC 19630 but sensitive to another WRN inhibitor, NSC 617145.

Conclusions

WRN is essential for survival of ATL cells. Our studies suggest that targeting the WRN helicase with small inhibitors is a novel promising strategy to target HTLV-1-transformed ATL cells.

     

Investigation on left ventricular multi-directional deformation in patients of hypertension with different LVEF

Background

This study is aimed at investigating myocardial multi-directional systolic deformation in hypertensive with different left ventricular ejection fraction (LVEF), and exploring its contribution to LVEF.

Methods

One hundred and twenty-three patients with primary hypertension (HT) were divided into group A (LVEF ≥ 55%), group B (45% ≤ LVEF < 50%, or 50% ≤ LVEF < 55% + LVEDVI ≥ 97 ml/m²), and group C (LVEF < 45%). Two-dimensional strain echocardiography (2DSE) including LV longitudinal strain (SL), radial strain (SR) and circumferential strain (SC) were measured.

Results

SL decreased gradually from group A, B to C (all p < 0.05) while SR and SC were reduced only in group B and C (all p < 0.05). All strain measurements correlated to LVEF, with the strongest correlation in SC (r = ?0.82, p < 0.01) and the second in SL (r = ?0.76). The diastolic E/e increased from group A, B to C.

Conclusions

Left ventricular multi-directional deformation correlated well to LVEF in hypertension and particularly SC, indicating that it was SC, not SL or SR, that makes the prominent contribution to left ventricular pump function.

     

A lost opportunity for science: journals promote data sharing in metabolomics but do not enforce it

Introduction

Data sharing is being increasingly required by journals and has been heralded as a solution to the ‘replication crisis’.

Objectives

(i) Review data sharing policies of journals publishing the most metabolomics papers associated with open data and (ii) compare these journals’ policies to those that publish the most metabolomics papers.

Methods

A PubMed search was used to identify metabolomics papers. Metabolomics data repositories were manually searched for linked publications.

Results

Journals that support data sharing are not necessarily those with the most papers associated to open metabolomics data.

Conclusion

Further efforts are required to improve data sharing in metabolomics.

     

Organizing life cycle management in practice: challenges of a multinational manufacturing corporation

Purpose

The environmental life cycle management (LCM) literature proposes many factors considered critical in order to successfully conduct LCM. This study contrasts these vague and general factors proposed as critical to LCM in existing literature, with detailed accounts of LCM in practice.

Methods

A literature review of three related research fields, i.e., LCM, life cycle thinking, and sustainable supply chain management, is contrasted with a study of how LCM is enacted in practice in a large multinational manufacturing company recognized for its LCM work. A qualitative study, with mainly a managerial focus, is conducted based on interviews, workshops, part-time observations, and document studies.

Results and discussion

The literature review demonstrates that the three related research fields provide different accounts of LCM: all apply a holistic environmental perspective, but with different emphases and using largely different research methods. The empirical study shows that integration was a common topic at the studied company and that solutions were often sought in tools and processes. Middle management support proved important, and challenging, in these integration efforts. Challenges identified also included further integrating LCM into departments such as purchasing and sales.

Conclusions

The constant focus on integration at the studied company implies that LCM work is an ongoing effort. Several integration paths are identified: (1) inclusion of sustainability aspects in tools and processes, (2) finding ways to work around certain organizational levels, and (3) using networks and social interaction to create commitment and integration. Although the concept of LCM implies a holistic approach, LCM in practice reveals a lack of a comprehensive overview of LCM-related initiatives and of involved sustainability practitioners within the studied organization.

     

Urine metabolic fingerprinting can be used to predict the risk of metritis and highlight the pathobiology of the disease in dairy cows

Introduction

Metritis is an uterine pathology that causes economic losses for the dairy industry. It is associated with lower reproductive efficiency, increased culling rates, decreased milk production and increased veterinary costs.

Objectives

To gain a more detailed view of the urine metabolome and to detect metabolite signature in cows with metritis. In addition, we aimed to identify early metabolites which can help to detect cows at risk to develop metritis in the future.

Methods

We used nuclear magnetic resonance spectroscopy starting at 8 and 4 weeks prior to the expected day of parturition, during the week of diagnosis of metritis, and at 4 and 8 weeks after diagnosis of metritis in Holstein dairy cows.

Results

At 8 weeks before parturition, pre-metritic cows had a total of 30 altered metabolites. Interestingly, 28 of them increased in urine when compared with control cows (P?<?0.05). At 4 weeks before parturition, 34 metabolites were altered. At the week of diagnosis of metritis a total of 20 metabolites were altered (P?<?0.05). The alteration continued at 4 and 8 weeks after diagnosis.

Conclusions

The metabolic fingerprints in the urine of pre-metritic and metritic cows point toward excretion of multiple amino acids, tricarboxylic acid cycle metabolites and monosaccharides. Combination of galactose, leucine, lysine and panthotenate at 8 weeks before parturition might serve as predictive biomarkers for metritis.

     

Application of residual polysaccharide-degrading enzymes in dried shiitake mushrooms as an enzyme preparation in food processing

Objective

To examine the activities of residual enzymes in dried shiitake mushrooms, which are a traditional foodstuff in Japanese cuisine, for possible applications in food processing.

Results

Polysaccharide-degrading enzymes remained intact in dried shiitake mushrooms and the activities of amylase, β-glucosidase and pectinase were high. A potato digestion was tested using dried shiitake powder. The enzymes reacted with potato tuber specimens to solubilize sugars even under a heterogeneous solid-state condition and that their reaction modes were different at 38 and 50 °C.

Conclusion

Dried shiitake mushrooms have a potential use in food processing as an enzyme preparation.

     

Effect of everolimus on the glucose metabolic pathway in mouse skeletal muscle cells (C2C12)

Introduction

Everolimus selectively inhibits mammalian target of rapamycin complex 1 (mTORC1) and exerts an antineoplastic effect. Metabolic disturbance has emerged as a common and unique side effect of everolimus.

Objectives

We used targeted metabolomic analysis to investigate the effects of everolimus on the intracellular glycometabolic pathway.

Methods

Mouse skeletal muscle cells (C2C12) were exposed to everolimus for 48 h, and changes in intracellular metabolites were determined by capillary electrophoresis time-of-flight mass spectrometry. mRNA abundance, protein expression and activity were measured for enzymes involved in glycometabolism and related pathways.

Results

Both extracellular and intracellular glucose levels increased with exposure to everolimus. Most intracellular glycometabolites were decreased by everolimus, including those involved in glycolysis and the pentose phosphate pathway, whereas no changes were observed in the tricarboxylic acid cycle. Everolimus suppressed mRNA expression of enzymes related to glycolysis, downstream of mTOR signaling enzymes and adenosine 5′-monophosphate protein kinases. The activity of key enzymes involved in glycolysis and the pentose phosphate pathway were decreased by everolimus. These results show that everolimus impairs glucose utilization in intracellular metabolism.

Conclusions

The present metabolomic analysis indicates that everolimus impairs glucose metabolism in muscle cells by lowering the activities of glycolysis and the pentose phosphate pathway.

     

Insulin inhibits the JNK mediated cell death via upregulation of AKT expression in Schwann cells grown in hyperglycemia

Background

Schwann cells (SCs) are the glial cells of the peripheral nervous system, which forms a thick insulating structure around the axons. Hyperglycemia is known physiologic conditions in both type I and type II diabetes which causes diabetic neuropathy. But the SC possesses insulin receptors even though glucose uptake is independent of insulin. Since the insulin level is highly altered in diabetes, it is of greater importance to evaluate their role in the Schwann cell survival and death.

Methods

Schwann cells were isolated from neonatal pups and grown with and without insulin in hyperglycemic medium to mimic diabetic condition for 24 and 48 h. We studied the cell viability using 3 (4,5-dimethylthiazol-2-yl) 2,5- diphenyltetrazolium bromide (MTT) and mitochondrial membrane potential (MMP) assay at different time interval on SCs. We also studied the protein and gene expression of Protein Kinase B (AKT) and Jun N-terminal kinase (JNK), which are greatly involved in cell survival and cell death respectively.

Results

The result shows that, high glucose levels for 48 h decrease the SC viability. Hyperglycemic condition induces the SC death by increasing the JNK expression which in turn reduces the MMP of glial cells. However, insulin administration for SCs grown in high glucose condition can reduce the JNK expression by activating AKT signaling pathway.

Conclusion

These observations demonstrate that the proper insulin balance is required for Schwann cells survival in hyperglycemic condition. Therefore, altered insulin signaling can be one of the reasons for demyelination of peripheral neurons in diabetic neuropathy.

     

The EcoSpold 2 format—why a new format?

Purpose

This paper introduces the new EcoSpold data format for life cycle inventory (LCI).

Methods

A short historical retrospect on data formats in the life cycle assessment (LCA) field is given. The guiding principles for the revision and implementation are explained. Some technical basics of the data format are described, and changes to the previous data format are explained.

Results

The EcoSpold 2 data format caters for new requirements that have arisen in the LCA field in recent years.

Conclusions

The new data format is the basis for the Ecoinvent v3 database, but since it is an open data format, it is expected to be adopted by other LCI databases. Several new concepts used in the new EcoSpold 2 data format open the way for new possibilities for the LCA practitioners and to expand the application of the datasets in other fields beyond LCA (e.g., Material Flow Analysis, Energy Balancing).

     

A methodology for elucidating regulatory mechanisms leading to changes in lipid profiles

Introduction

It is difficult to elucidate the metabolic and regulatory factors causing lipidome perturbations.

Objectives

This work simplifies this process.

Methods

A method has been developed to query an online holistic lipid metabolic network (of 7923 metabolites) to extract the pathways that connect the input list of lipids.

Results

The output enables pathway visualisation and the querying of other databases to identify potential regulators. When used to a study a plasma lipidome dataset of polycystic ovary syndrome, 14 enzymes were identified, of which 3 are linked to ELAVL1—an mRNA stabiliser.

Conclusion

This method provides a simplified approach to identifying potential regulators causing lipid-profile perturbations.

     

Untargeted and stable isotope-assisted metabolomic analysis of MDA-MB-231 cells under hypoxia

Introduction

Hypoxia commonly occurs in cancers and is highly related with the occurrence, development and metastasis of cancer. Treatment of triple negative breast cancer remains challenge. Knowledge about the metabolic status of triple negative breast cancer cell lines in hypoxia is valuable for the understanding of molecular mechanisms of this tumor subtype to develop effective therapeutics.

Objectives

Comprehensively characterize the metabolic profiles of triple negative breast cancer cell line MDA-MB-231 in normoxia and hypoxia and the pathways involved in metabolic changes in hypoxia.

Methods

Differences in metabolic profiles affected pathways of MDA-MB-231 cells in normoxia and hypoxia were characterized using GC–MS based untargeted and stable isotope assisted metabolomic techniques.

Results

Thirty-three metabolites were significantly changed in hypoxia and nine pathways were involved. Hypoxia increased glycolysis, inhibited TCA cycle, pentose phosphate pathway and pyruvate carboxylation, while increased glutaminolysis in MDA-MB-231 cells.

Conclusion

The current results provide metabolic differences of MDA-MB-231 cells in normoxia and hypoxia conditions as well as the involved metabolic pathways, demonstrating the power of combined use of untargeted and stable isotope-assisted metabolomic methods in comprehensive metabolomic analysis.

     

Life cycle management of energy-consuming products in companies using IO-LCA

Background, aim, and scope

Today, the effective integration of life cycle thinking into existing business routines is argued to be the most critical step for more sustainable business models. The study tests the suitability of an input–output life cycle assessment (IO-LCA) approach in screening life cycle impacts of energy-using products in companies. It estimates the life cycle impacts of three products and assesses the suitability of such approach in a company environment.

Materials and methods

The multiple case studies evaluate the suitability of an IO-LCA method in a company environment. A comprehensive life cycle cost and impact study of three product systems (building ventilation system, information and communication technology (ICT) network product, and welding machine) is conducted and the life cycle phases with highest economical and environmental contribution are determined. Scenario analysis is performed in order to assess the sensitivity of the results to major changes in the studied systems. Finally, the usability of the IO-LCA approach for environmental evaluations in companies is assessed by collecting data on workload and interviewing the participating workers and managers.

Results

The results showed that the use phase with operating energy was environmentally important in all evaluated energy-using products. However, only in one case (ICT network product) the use was the single most significant life cycle phase. In two other cases, the sourcing was equally important. The results also indicated that the IO-LCA approach is much easier to adapt by current management of companies because it automatically links life cycle costs to environmental indicators and, by order of magnitude, reduces the workload in companies.

Discussion

It appears that the IO-LCA approach can be used to screen environmentally significant life cycle phases of energy-using products in companies by utilizing readily available accounting or other documented data. The IO-LCA approach produced comparable results with the ones published in traditional process-based LCA literature. In addition to the main results, some practical benefits of using the IO-LCA could also be suggested: the approach was very fast to use and would thus allow an easier adoption of environmental evaluations in companies as well as wider environmental testing of products in early conceptual design phase.

Conclusions

The results indicated that the IO-LCA approach could clearly offer added value to the environmental management of companies. The IO-LCA was found to provide a very fast access to the key life cycle characteristics of products. Similarly, it offered practical means to integrate life cycle thinking into existing business routines and to activate the decision makers in companies by giving them easily comprehendible results.

Recommendations and perspectives

The results would suggest that similar environmental IO tables, besides the US ones used here, would have value and should be collected for other major geographical and economical regions. The tables would enable a much larger share of companies to manage their environmental issues. It also seems that, because the user profile is so dominant in the case of energy-using products, more studies, both theoretical (How to valuate the future behavior in environmental studies?) and empirical (What really creates value for users?), should focus on the behavior of users.