Application of next-generation sequencing methods for microbial monitoring of anaerobic digestion of lignocellulosic biomass

The anaerobic digestion of lignocellulosic wastes is considered an efficient method for managing the world’s energy shortages and resolving contemporary environmental problems. However, the recalcitrance of lignocellulosic biomass represents a barrier to maximizing biogas production. The purpose of this review is to examine the extent to which sequencing methods can be employed to monitor such biofuel conversion processes. From a microbial perspective, we present a detailed insight into anaerobic digesters that utilize lignocellulosic biomass and discuss some benefits and disadvantages associated with the microbial sequencing techniques that are typically applied. We further evaluate the extent to which a hybrid approach incorporating a variation of existing methods can be utilized to develop a more in-depth understanding of microbial communities. It is hoped that this deeper knowledge will enhance the reliability and extent of research findings with the end objective of improving the stability of anaerobic digesters that manage lignocellulosic biomass.     

Characterization of Molecular Determinants of the Conformational Stability of Macrophage Migration Inhibitory Factor: Leucine 46 Hydrophobic Pocket

Macrophage Migration Inhibitory Factor (MIF) is a key mediator of inflammatory responses and innate immunity and has been implicated in the pathogenesis of several inflammatory and autoimmune diseases. The oligomerization of MIF, more specifically trimer formation, is essential for its keto-enol tautomerase activity and probably mediates several of its interactions and biological activities, including its binding to its receptor CD74 and activation of certain signaling pathways. Therefore, understanding the molecular factors governing the oligomerization of MIF and the role of quaternary structure in modulating its structural stability and multifunctional properties is crucial for understanding the function of MIF in health and disease. Herein, we describe highly conserved intersubunit interactions involving the hydrophobic packing of the side chain of Leu46 onto the β-strand β3 of one monomer within a hydrophobic pocket from the adjacent monomer constituted by residues Arg11, Val14, Phe18, Leu19, Val39, His40, Val41, Val42, and Pro43. To elucidate the structural significance of these intersubunit interactions and their relative contribution to MIF’s trimerization, structural stability and catalytic activity, we generated three point mutations where Leu46 was replaced by glycine (L46G), alanine (L46A) and phenylalanine (L46F), and their structural properties, stability, oligomerization state, and catalytic activity were characterized using a battery of biophysical methods and X-ray crystallography. Our findings provide new insights into the role of the Leu46 hydrophobic pocket in stabilizing the conformational state of MIF in solution. Disrupting the Leu46 hydrophobic interaction perturbs the secondary and tertiary structure of the protein but has no effect on its oligomerization state.     

The effects of sex and season on the metal levels and proximate composition of red mullet (Mullus barbatus Linnaeus 1758) caught from the Middle Black Sea

The effects of seasonal and sexual differences on the levels of macro, trace element, and toxic metal levels in Mullus barbatus were investigated using the ICP-MS. The quantity relationships of macro elements were found as K>P>Na>Ca>Mg for male, P>K>Na>Ca>Mg in female; the quantity relationships of trace elements were determined as Fe>Zn>Mn>Mo>Se>Cu>Ni>Co in male, Fe>Zn>Mn>Se>Cu>Ni>Mo>Co in female; and the quantity relationship of toxic elements were found as As>Pb>Cd in both male and female. Macro, trace, and toxic metal levels showed no significant differences (p > .05) in terms of sexuality, but demonstrated seasonal differences (p > .05). Overall, the highest level of macro elements was determined in the autumn, while the high level of trace elements was measured in the winter. The As, Cd, and Pb levels in the muscle tissue of red mullet fish were higher than acceptable limits. However, when the Estimated Daily Intake (EDI) and Estimated Weekly Intake (EWI) of both toxic metals are considered, they appear to be below acceptable limits. As a result, it was determined that the red mullet had been contaminated with toxic metals (Cd and Pb) but their levels according to EDI and EWI were below the limit.     

Stereoselective synthesis of new rac-quercitols containing hydroxymethyl groups as glucosidase inhibitors

Stereoselective and efficient synthesis of hydroxymethyl-substituted rac-quercitols (13–15) was achieved, starting from cis-furan (Kobayashi, 2008) with photooxygenation reaction, which is readily available by the reduction of cis-phtalic anhydride. α- and β-Glucosidase enzyme activity of the target molecules was evaluated and good inhibitor activity was seen. One- and two-dimensional NMR spectroscopy, IR spectroscopy and X-ray crystallography were utilized in the structure characterization of products.     

Implementation of fed-batch strategies for vitamin K (menaquinone-7) production by Bacillus subtilis natto in biofilm reactors

Recent studies show the essential health benefits associated with vitamin K, especially menaquinone-7 (MK-7). These benefits include reducing risks of cardiovascular diseases, osteoporosis, and even cancer. However, MK-7 production on an industrial level is only possible through bacterial fermentation and also current static fermentation strategies are not potent enough with difficulties to scale up. Biofilm reactors, however, may be a practical alternative. Biofilm reactors provide a controlled environment for the microorganisms to form mature and robust biofilms that enable them to produce value-added products with enhanced efficiencies. In this study, fed-batch addition of glucose and glycerol were investigated to the base media in biofilm reactors, as carbon source addition seemed crucial in batch fermentations. Results indicated that fed-batch strategies can be significantly effective in glucose-based medium, increasing the end-product concentrations to 28.7 ± 0.3 mg/L of MK-7 which was 2.3 fold higher than the level produced in suspended-cell bioreactors and renders the biofilm reactors as a potential replacement for static fermentation strategies. Moreover, morphological changes of B. subtilis were tracked during the 12-day long fermentation runs and finally, SEM investigations confirmed significant biofilm and extracellular matrices formed on the plastic composite support (PCS) in the biofilm reactors. In conclusion, biofilm reactors especially with fed-batch fermentation regimes seem to be an effective tool for MK-7 production at industrial scales.

     

Specific localization of the respiratory alternative oxidase in meristematic and xylematic tissues from developing soybean roots and hypocotyls.

We used tissue printing and specific immunostaining to examine the localization of the alternative oxidase (AOX) protein in correlation with measurements of AOX capacity. Selected root and hypocotyl regions were analyzed during the first 14 d of growth. It is shown that AOX protein is localized in the apical meristem and in developing xylem. The temporal pattern of expression is coincident with the evolution of AOX capacity. Data suggest that AOX expression is linked to xylem differentiation. Since heat is a major product of the alternative pathway, we speculate that thermogenesis is implicated in morphogenesis.     

Plasma glutathione peroxidase activity and selenium levels of newborns with jaundice

The plasma glutathione peroxidase (GSH-Px) and selenium (Se) levels were determined in 31 newborns affected by jaundice (NWJ). The GSH-Px levels of both full-term and premature newborns exhibiting jaundice and having a birthweight lower than 2000 g were significantly low (p<0.05) when compared to controls. No significant differences were found in the corresponding Se levels, which were similar in all groups and independent of the pregnancy period and birthweight.     

A chemical and biological approach towards the definition of calcareous soils

Summary Several agricultural problems are associated with the presence of certain levels of CaCO₃ in soils. The level of CaCO₃ at which the phosphate fixation becomes an apparent agricultural problem, is thought to be an appropriate margine at which the soil can be considered calcareous. Thus, a set of soil mixtures, ranging in CaCO₃ content from 1 to 96% was prepared and used in a column study to determine the level at which the CaCO₃ fraction becomes a dominant factor controlling. P³² movement and distribution.Increasing the percentage of oolitic sand, in the soil mixture, from 1 to 10% caused a sharp drop in P³² movement with soil solution and any increase in CaCO₃ content above 10% did not show any further drop in P³² movement. The amount of P³² removed with the soil solution was generally low compared to that retained in soil columns. Studying the distribution of P³² in soil columns, after five displacements, has indicated that the migration of P³² from the top soil increased by increasing CaCO₃ from 1 and 2 to 6%. The amount of P³² removed was however retained in lower sections. A very sharp decrease in P³² migration from the top soil was observed when CaCO₃ content was raised from 8 to 10%.A similar picture was shown when the CaCO₃ material used was in clay size fraction. However the sharp increase in phosphate retention in top soil sections took place at CaCO₃ content of 8% rather than at 10%. A limit of 8 to 10% CaCO₃ was proposed as an appropriate margine for defining calcareous soils.