Dynamics of the KB Proton Pathway in Cytochrome ba3 from Thermus thermophilus

The ba₃ cytochrome c oxidase from Thermus thermophilus is a B-type oxygen-reducing heme-copper oxidase and a proton pump. It uses only one proton pathway for transfer of protons to the catalytic site, the K^B pathway. It was previously shown that the ba₃ oxidase has an overall similar reaction sequence to that in mitochondrial-like A-type oxidases. However, the timing of loading the pump site, and formation and decay of catalytic intermediates is different in the two types of oxidases. In the present study, we have investigated variants in which two amino acids of the K^B proton pathway leading to the catalytic site were exchanged; Tyr-248 (located ∼23 Å below the active site towards the cytoplasm) in subunit I (Y248T) and Glu-15 (∼26 Å below the active site, ∼16 Å from Tyr-248) in subunit II (E15^IIQ). Even though the overall catalytic turnover in these two variants is similar and very low (<1 % of wildtype), the substitutions had distinctly different effects on the kinetics of proton transfer to the catalytic site. The results indicate that the Glu-15^II is the only essentially crucial residue of the K^B pathway, but that the Tyr-248 also plays a distinct role in defining an internal proton donor and controlling the dynamics of proton transfer to the pump site and the catalytic site.     

Docosahexaenoic Acid Supplementation during Pregnancy: A Potential Tool to Prevent Membrane Rupture and Preterm Labor

Polyunsaturated fatty acids (PUFAs) are required to maintain the fluidity, permeability and integrity of cell membranes. Maternal dietary supplementation with ω-3 PUFAs during pregnancy has beneficial effects, including increased gestational length and reduced risk of pregnancy complications. Significant amounts of ω-3 docosahexaenoic acid (DHA) are transferred from maternal to fetal blood, hence ensuring high levels of DHA in the placenta and fetal bloodstream and tissues. Fetal DHA demand increases exponentially with gestational age, especially in the third trimester, due to fetal development. According to the World Health Organization (WHO) and the Food and Agriculture Organization of the United Nations (FAO), a daily intake of DHA is recommended during pregnancy. Omega-3 PUFAs are involved in several anti-inflammatory, pro-resolving and anti-oxidative pathways. Several placental disorders, such as intrauterine growth restriction, premature rupture of membranes (PROM) and preterm-PROM (pPROM), are associated with placental inflammation and oxidative stress. This pilot study reports on a preliminary evaluation of the significance of the daily DHA administration on PROM and pPROM events in healthy pregnant women. Further extensive clinical trials will be necessary to fully elucidate the correlation between DHA administration during pregnancy and PROM/pPROM occurrence, which is related in turn to gestational duration and overall fetal health.     

Quantitative Evaluation of the Pyruvic Acid Content in Onion Samples with a Fully Validated High-Performance Liquid Chromatography Method

Onion cultivars with low pungency have been increasing in popularity because they are more attractive for fresh, uncooked use. The pyruvic acid amount is commonly used as a measure of onion pungency. For the first time the validation of a reversed-phase high-performance liquid chromatography method for the quantitative analysis of pyruvic acid in onion extracts was carried out. The established chromatography method based on the use of a 50 mM phosphate buffer (pH 2.5) eluent system provided good precision (RSD% in the range of 1.25–1.29, in the long-period) and accuracy (recovery% in the range of 97.70–111.62, in the long-period) as well as appreciably low limit of detection and limit of quantitation values (11.03 and 33.41 ng/mL, respectively). The validated method proved successful in the analysis of the onion extract from Dorata di Parma cultivar after a preliminary solid-phase extraction step. A significant adherence with results from previous spectrophotometric determinations turned out.     

Fuzzy PWM-PID control of cocontracting antagonistic shape memory alloy muscle pairs in an artificial finger

This paper presents biomimetic control of an anthropomorphic artificial finger actuated by three antagonistic shape memory alloy (SMA) muscle pairs that are each configured in a dual spring-biased configuration. This actuation system forms the basis for biomimetic tendon-driven flexion/extension of the metacarpophalangeal (MCP) and proximal interphalangeal (PIP) joints of the artificial finger, as well as the abduction/adduction of its MCP joint. This work focuses on the design and experimental verification of a new fuzzy pulse-width-modulated proportional-integral-derivative (i.e. fuzzy PWM-PID) controller that is capable of realizing cocontraction of the SMA muscle pairs, as well as online tuning of the PID gains to deal with system nonlinearities and parameter uncertainties. The main contribution of this paper is the proposed biomimetic cocontraction control strategy, which co-activates the antagonistic muscle pairs as a synergistic functional unit. It emulates a similar strategy in neural control, called “common drive,” employed by the central nervous system (CNS). In order to maintain a desired position of a joint, the corresponding agonistic muscle pairs are cocontracted by the CNS and stiffen the joint. The synthesis and parametric analysis of the proposed controller are carried out via numerical simulations using a dynamic model of the system. The performance advantage of the cocontracting fuzzy PWM-PID controller over the original PWM-PID controller is shown by both numerical and experimental results. A successful application of the new controller to fingertip trajectory tracking tasks using the MCP joint’s flexion/extension and abduction/adduction is also described.     

Enhanced Production of Bioactive Isoprenoid Compounds from Cell Suspension Cultures of Artemisia annua L. Using β-Cyclodextrins

Plant cell cultures as valuable tools for the production of specific metabolites can be greatly improved by the application of elicitors including cyclodextrins (CDs) for enhancing the yields of the desired plant compounds. Here the effects of 2,6-dimethyl-β-cyclodextrins (DIMEB) on the production of carotenoids and quinones from Artemisia annua L. cell suspension cultures were investigated. The addition of 50 mM DIMEB induced an early increase of intracellular carotenoid and quinone contents, which could be observed to a higher extent for lutein (10-fold), Q9 (3-fold) and Q10 (2.5-fold). Real Time PCR analysis revealed that the expression of 1-deoxy-d-xylulose-5-phosphate reductoisomerase (DXR) gene in DIMEB treated cell cultures after three days was 2.5-fold higher than in untreated samples, thus suggesting that the DIMEB induced increase of carotenoids and quinones could be due to the induction of the plastidial isoprenoid biosynthetic route. In addition, the DIMEB treatment induced an enhanced release of carotenoids and quinones into the culture medium of A. annua cell suspension cultures possibly due to the ability of CDs to form inclusion complexes with hydrophobic molecules.     

Meta-Omic Platforms to Assist in the Understanding of NAFLD Gut Microbiota Alterations: Tools and Applications

Non-alcoholic fatty liver disease (NAFLD) is the most common cause of chronic liver disease worldwide as a result of the increasing prevalence of obesity, starting from early life stages. It is characterized by a spectrum of liver diseases ranging from simple fatty liver (NAFL) to steatohepatitis (NASH), with a possible progression to fibrosis, thus increasing liver-related morbidity and mortality. NAFLD development is driven by the co-action of several risk factors, including obesity and metabolic syndrome, which may be both genetically induced and diet-related. Recently, particular attention has been paid to the gut-liver axis, which may play a physio-pathological role in the onset and progression of the disease. The gut microbiota is intended to act as a bioreactor that can guarantee autonomous metabolic and immunological functions and that can drive functional strategies within the environment of the body in response to external stimuli. The complexity of the gut microbiota suggests that it behaves as an organ. Therefore, the concept of the gut-liver axis must be complemented with the gut-microbiota-liver network due to the high intricacy of the microbiota components and metabolic activities; these activities form the active diet-driven power plant of the host. Such complexity can only be revealed using systems biology, which can integrate clinical phenomics and gut microbiota data.