Light dazzles from the black box: Whole-cell biosensors are ready to inform on fundamental soil biological processes

Whole-cell biosensors are natural or engineered microorganisms producing signals in response to specific stimuli. This review introduces the use of whole-cell biosensors for the study of the soil system, discuss the recent developments and some current limitations and draws future prospects of the whole-cell biosensors for application to the study of the agro-ecosystems. The review focuses mainly on the lux- and gfp-inserted whole-cell biosensors producing bioluminescence and multicoloured fluorescent proteins, which allow an easy and reproducible detection of the signals from a large number of prokaryotic and eukaryotic soil-borne microorganisms. This review also points out how the whole-cell biosensors indicate the bioavailability of selected analyte, an information that cannot be straight forwardly extrapolated using the chemical methods of soil analysis. However, regardless of the immense progress in biotechnology and genetics that allows to construct whole-cell biosensors for virtually detecting any chemical at ultra low concentrations, the soil still remains the most extreme natural system to be studied with these biotechnological analytical tools. Although a lack of standardization for most of the constructed whole-cell biosensors along with the scarce knowledge of their performance concur to prevent their use in the official methods of soil and environmental analysis, owing to their stability and selectivity we restate that the whole-cell biosensors are ready to provide information on the main processes occurring in soil, and represent unprecedented sensitive tools for improving agriculture and for soil monitorin

Is soluble protein mineralisation and protease activity in soil regulated by supply or demand?

Protein represents a major input of organic matter to soil and is an important source of carbon (C) and nitrogen (N) for microorganisms. Therefore, determining which soil properties influence protein mineralisation in soil is key to understanding and modelling soil C and N cycling. However, the effect of different soil properties on protein mineralisation, and especially the interactions between soil properties, are poorly understood. We investigated how topsoil and subsoil properties affect protein mineralisation along a grassland altitudinal (catena) sequence that contained a gradient in soil type and primary productivity. We devised a schematic diagram to test the key edaphic factors that may influence protein mineralisation in soil (e.g. pH, microbial biomass, inorganic and organic N availability, enzyme activity and sorption). We then measured the mineralisation rate of 14C-labelled soluble plant-derived protein and amino acids in soil over a two-month period. Correlation analysis was used to determine the associations between rates of protein mineralisation and soil properties. Contrary to expectation, we found that protein mineralisation rate was nearly as fast as for amino acid turnover. We ascribe this rapid protein turnover to the low levels of protein used here, its soluble nature, a high degree of functional redundancy in the microbial community and microbial enzyme adaptation to their ecological niche. Unlike other key soil N processes (e.g. nitrification, denitrification), protease activity was not regulated by a small range of factors, but rather appeared to be affected by a wide range of interacting factors whose importance was dependent on altitude and soil depth [e.g. above-ground net primary productivity (NPP), soil pH, nitrate, cation exchange capacity (CEC), C:N ratio]. Based on our results, we hypothesise that differences in soil N cycling and the generation of ammonium are more related to the rate of protein supply rather than limitations in protease activity and protein turnover per se

Non-invasive cardiac imaging techniques and vascular tools for the assessment of cardiovascular disease in type 2 diabetes mellitus

Cardiovascular disease is the major cause of mortality in type 2 diabetes mellitus. The criteria for the selection of those asymptomatic patients with type 2 diabetes who should undergo cardiac screening and the therapeutic consequences of screening remain controversial. Non-invasive techniques as markers of atherosclerosis and myocardial ischaemia may aid risk stratification and the implementation of tailored therapy for the patient with type 2 diabetes. In the present article we review the literature on the implementation of non-invasive vascular tools and cardiac imaging techniques in this patient group. The value of these techniques as endpoints in clinical trials and as risk estimators in asymptomatic diabetic patients is discussed. Carotid intima–media thickness, arterial stiffness and flow-mediated dilation are abnormal long before the onset of type 2 diabetes. These vascular tools are therefore most likely to be useful for the identification of ‘at risk’ patients during the early stages of atherosclerotic disease. The additional value of these tools in risk stratification and tailored therapy in type 2 diabetes remains to be proven. Cardiac imaging techniques are more justified in individuals with a strong clinical suspicion of advanced coronary heart disease (CHD). Asymptomatic myocardial ischaemia can be detected by stress echocardiography and myocardial perfusion imaging. The more recently developed non-invasive multi-slice computed tomography angiography is recommended for exclusion of CHD, and can therefore be used to screen asymptomatic patients with type 2 diabetes, but has the associated disadvantages of high radiation exposure and costs. Therefore, we propose an algorithm for the screening of asymptomatic diabetic patients, the first step of which consists of coronary artery calcium score assessment and exercise ECG

Effects of Anacetrapib in Patients with Atherosclerotic Vascular Disease

BACKGROUND: Patients with atherosclerotic vascular disease remain at high risk for cardiovascular events despite effective statin-based treatment of low-density lipoprotein (LDL) cholesterol levels. The inhibition of cholesteryl ester transfer protein (CETP) by anacetrapib reduces LDL cholesterol levels and increases high-density lipoprotein (HDL) cholesterol levels. However, trials of other CETP inhibitors have shown neutral or adverse effects on cardiovascular outcomes. METHODS: We conducted a randomized, double-blind, placebo-controlled trial involving 30,449 adults with atherosclerotic vascular disease who were receiving intensive atorvastatin therapy and who had a mean LDL cholesterol level of 61 mg per deciliter (1.58 mmol per liter), a mean non-HDL cholesterol level of 92 mg per deciliter (2.38 mmol per liter), and a mean HDL cholesterol level of 40 mg per deciliter (1.03 mmol per liter). The patients were assigned to receive either 100 mg of anacetrapib once daily (15,225 patients) or matching placebo (15,224 patients). The primary outcome was the first major coronary event, a composite of coronary death, myocardial infarction, or coronary revascularization. RESULTS: During the median follow-up period of 4.1 years, the primary outcome occurred in significantly fewer patients in the anacetrapib group than in the placebo group (1640 of 15,225 patients [10.8%] vs. 1803 of 15,224 patients [11.8%]; rate ratio, 0.91; 95% confidence interval, 0.85 to 0.97; P=0.004). The relative difference in risk was similar across multiple prespecified subgroups. At the trial midpoint, the mean level of HDL cholesterol was higher by 43 mg per deciliter (1.12 mmol per liter) in the anacetrapib group than in the placebo group (a relative difference of 104%), and the mean level of non-HDL cholesterol was lower by 17 mg per deciliter (0.44 mmol per liter), a relative difference of -18%. There were no significant between-group differences in the risk of death, cancer, or other serious adverse events. CONCLUSIONS: Among patients with atherosclerotic vascular disease who were receiving intensive statin therapy, the use of anacetrapib resulted in a lower incidence of major coronary events than the use of placebo. (Funded by Merck and others; Current Controlled Trials number, ISRCTN48678192 ; ClinicalTrials.gov number, NCT01252953 ; and EudraCT number, 2010-023467-18 .)

Effects of Biochar on the C Use Efficiency of Soil Microbial Communities: Components and Mechanisms

Biochar production and incorporation into soil is gaining momentum as a sustainable strategy for climate change mitigation, supported by ever increasing reports of significant carbon (C) sequestration in soil and reduction in greenhouse gas (GHG) emissions from the amended soils. With the progression in biochar testing and use, there is also emerging evidence that biochar induces C sequestration in soil, and that it may not be solely caused by its inherent chemical stability, but also by the complex microbially driven processes and an increase in C use efficiency (CUE) through soil microbial metabolism. This evidence contradicts the current paradigm that sees the microbial CUE decrease during the degradation of recalcitrant material due to thermodynamic constraints, as observed only in several short-term and pilot-scale trials. As the CUE in soil results from interactions between several abiotic and biotic factors, in this paper we examine the link between the biochar properties, soil physico-chemical properties and microbial physiology to explain the CUE increase reported for biochar-amended soils. Based on the large body of physico-chemical literature, and on the high functional diversity and metabolic flexibility of soil microbial communities, we hypothesize that the long-term stabilization of biochar-borne C in the soil systems is not only controlled by its inherent recalcitrance, but also by the cooperative actions of improved soil status and increased microbial CUE. Given that the current knowledge on this specific aspect is still poor, in this feature paper we summarize the state of knowledge and examine the potential impact of biochar on some factors contributing to the whole-soil CUE. We conclude that, beside its inherent recalcitrance, biochar weathering and oxidation in soil create physical and chemical conditions that can potentially increase the microbial CUE. While these processes stabilize the microbial processed C in soil and increase soil fertility, more data from long-term field trials are needed to model the relationship between the CUE and the MRT of biochar-borne C. Based on our hypotheses and relying upon analysis of the available literature, we also suggest possible research approaches that may contribute to filling the gaps in the current knowledge on the topic

Activities of proteolytic enzymes

Proteases, also known as proteinases or proteolytic enzymes, are a large group of hydrolases that catalyze the cleavage of peptide bonds in proteins to produce peptides and/or amino acids. Classification of proteolytic enzymes is based on three major criteria: type of reaction catalyzed, functional group of the active site, and type of molecular structure and evolutionary relationship among the various enzymes. According to the Nomenclature Committee of the International Union of Biochemistry and Molecular Biology, the proteolytic enzymes can be grouped into proteases and peptidases on the basis of their nature of attack. This chapter describes methods to estimate soil protease activity, utilizing two substrates: casein, essentially a nonspecific substrate, and N-benzoyl-L-argininamide (BAA), a typical substrate for trypsin-like enzyme. The assay is based on colorimetric estimation of products released by the protein and amide-hydrolyzing enzymes when soil is incubated with buffered solutions of casein and BAA, respectively

Enzymes activities, detection and expression of genes codyfing enzymes

Enzymes activities, detection and expression of genes codyfing enzymes

Nutrizione parenterale in rianimazione. Studio di 190 casi

Results observed in 190 patients receiving parenteral nutrition (including 104 thus treated for periods of 3 to 28 days) are presented. The data point to the importance of both the quantity and quality of the calorie and nitrogen intake. The energy and plastic sources used in the treatment are discussed. The modalities most suited for employment in patients with unimpaired renal functional and in those with serious renal insufficiency are described in the light of personal experience