Sepsis represents a deranged and exaggerated systemic inflammatory response to infection and is associated with vascular and metabolic abnormalities that trigger systemic organic dysfunction. Mitochondrial function has been shown to be severely impaired during the early phase of critical illness, with a reduction in biogenesis, increased generation of reactive oxygen species and a decrease in adenosine triphosphate synthesis of up to 50%. Mitochondrial dysfunction can be assessed using mitochondrial DNA concentration and respirometry assays, particularly in peripheral mononuclear cells. Isolation of monocytes and lymphocytes seems to be the most promising strategy for measuring mitochondrial activity in clinical settings because of the ease of collection, sample processing, and clinical relevance of the association between metabolic alterations and deficient immune responses in mononuclear cells. Studies have reported alterations in these variables in patients with sepsis compared with healthy controls and non-septic patients. However, few studies have explored the association between mitochondrial dysfunction in immune mononuclear cells and unfavorable clinical outcomes. An improvement in mitochondrial parameters in sepsis could theoretically serve as a biomarker of clinical recovery and response to oxygen and vasopressor therapies as well as reveal unexplored pathophysiological mechanistic targets. These features highlight the need for further studies on mitochondrial metabolism in immune cells as a feasible tool to evaluate patients in intensive care settings. The evaluation of mitochondrial metabolism is a promising tool for the evaluation and management of critically ill patients, especially those with sepsis. In this article, we explore the pathophysiological aspects, main methods of measurement, and the main studies in this field.

Respirometry tests are a widely employed method in wastewater treatment field to characterize wastewater streams, assess toxic/inhibitory effects to the biomass, calibrate mathematical models. Respirometry can allow to fractionize the chemical oxygen demand (COD) in biodegradable and inert fractions, but also provide information related to biomass kinetics and stoichiometry through standardized laboratory techniques. Considering the increasing number of emerging contaminants detected in wastewater effluents, such as pharmaceuticals, personal care products and pesticides, respirometry can be a useful tool to promptly assess any toxic or inhibitory effect in wastewater treatment plants (WWTPs) operations. Beside conventional activated sludge (CAS), in recent years respirometric methods have been applied to innovative fields, such as moving-bed bio-reactors (MBBRs), fungi and microalgae, exploiting natural remediation methods. In particular, respirometry application to microalgae, through the so-called photo-respirometry, has been investigated in the latest years in the treatment of high-nutrient loaded streams, allowing resource recovery in biomass form. In this work, respirometric methods are first introduced from a theoretical basis and then critically discussed by considering the experimental apparatus, the available characterization protocols and the fields of application; the most recent literature findings on respirometry are coupled with authors\' experience in the field. A comparison between physicochemical methods and respirometry is made, considering common protocols for WWTP modelling and calibration. The future research needed on the topic is finally outlined, including the coupling of respirometry with microbial community analysis, potentially leading to an enhanced process understanding, an extended respirometry utilization to get specific kinetic and stoichiometric parameters for modelling purposes, and a wider respirometry application as diagnosis tool in WWTP operations.

Oxidation of fatty acids is a major source of energy in the heart, liver, and skeletal muscle. It can be measured accurately using respirometry in isolated mitochondria, intact cells, and permeabilized cells or tissues. This technique directly measures the rate of oxygen consumption or flux at various respiratory states when appropriate substrates, uncouplers, and inhibitors are used. Acylcarnitines such as palmitoylcarnitine or octanoylcarnitine are the commonly used substrates. The β-oxidation pathway is prone to feedforward inhibition resulting from accumulation of short-chain acyl-CoA and depletion of CoA, but inclusion of malate or carnitine prevents accumulation of these intermediaries and CoA depletion.