Structure of YdjH from Acinetobacter baumannii revealed an active site of YdjH family sugar kinase.

Bacterial sugar kinase is a central enzyme for proper sugar degradation in bacteria, essential for survival and growth. Therefore, this enzyme family is a primary target for antibacterial drug development, with YdjH most preferring to phosphorylate higher-order monosaccharides with a carboxylate terminus. Sugar kinases express diverse specificity and functions, making specificity determination of this family a prominent issue. This study examines the YdjH crystal structure from Acinetobacter baumannii (abYdjH), which has an exceptionally high antibiotic resistance and is considered a superbug. Our structural and biochemical study revealed that abYdjH has a widely open lid domain and is a solution dimer. In addition, the putative active site of abYdjH was determined based on structural analysis, sequence comparison, and in silico docking. Finally, we proposed the active site-forming residues that determine various sugar specificities from abYdjH. This study contributes towards a deeper understanding of the phosphorylation process and bacterial sugar metabolism of YdjH family to design the next-generation antibiotics for targeting A. baumannii.

The structure of MucD from Pseudomonas syringae revealed N-terminal loop-mediated trimerization of HtrA-like serine protease.

Protein quality control mechanisms are essential for maintaining cellular integrity, and the HtrA family of serine proteases plays a crucial role in handling folding stress in prokaryotic periplasm. Escherichia coli harbors three HtrA members, namely, DegS, DegP, and DegQ, which share a common domain structure. MucD, a putative HtrA family member that resembles DegP, is involved in alginate biosynthesis regulation and the stress response. Pseudomonas syringae causes plant diseases and opportunistic infections in humans. This study presents the high-resolution structure of MucD from Pseudomonas syringae (psMucD), revealing its composition as a typical HtrA family serine protease with protease and PDZ domains. Its findings suggest that psMucD containing one PDZ domain is a trimer in solution, and psMucD trimerization is mediated by its N-terminal loop. Sequence and structural analyses revealed similarities and differences with other HtrA family members. Additionally, this study provides a model of psMucD's catalytic process, comparing it with other members of the HtrA family of serine proteases.

Effects of Melting Conditions on Cerium Oxidation State and Catalytic Properties of CeO2-P2O5 Glass Systems.

As with any solvent, stabilizing a multivalent element at a given oxidation state in glass depends on the thermodynamic conditions. The effects of temperature on the oxidation-reduction equilibrium have been previously noted with higher temperatures being more conducive to reduced states. Herein, 30CeO2-70P2O5 binary system glasses were prepared. The melting temperature and time dependency on Ce4+ and Ce3+ ion concentrations were studied using X-ray photoelectron spectroscopy. Different melting conditions were investigated at temperatures ranging from 1300 °C to 1500 °C for 60 min, and at 1400 °C for durations ranging from 30 min to 90 min. The changes in the catalytic properties of the glasses as a function of Ce4+ and Ce3+ ion concentrations were confirmed based on the changes in the decomposition starting temperatures using thermogravimetric analysis. The main changes in the oxidation states according to melting conditions were discussed in terms of the catalytic properties of CeO2-P2O5 glass systems.

Impact of commercial cigarette smoke condensate on brain tissue co-cultured with astrocytes and blood-brain barrier endothelial cells.

The purpose of the current study was to investigate the effect of two commercial cigarette smoke condensates (CCSC) on oxidative stress and cell cytotoxicity in human brain (T98G) or astrocytes (U-373 MG) in the presence of human brain microvascular endothelial cells (HBMEC). Cell viability of mono-culture of T98G or U-373 MG was markedly decreased in a concentration-dependent manner, and T98G was more susceptible than U-373 MG to CCSC exposure. Cytotoxicity was less prominent when T98G was co-cultured with HBMEC than when T98G was co-cultured with U-373 MG. Significant reduction in trans-epithelial electric resistance (TEER), a biomarker of cellular integrity was noted in HBMEC co-cultured with T98G (HBMEC-T98G co-culture) and U-373 MG co-cultured with T98G (U-373 MG-T98G co-culture) after 24 or 48 hr CCSC exposure, respectively. TEER value of U-373 MG co-cultured with T98G (79-84%) was higher than HBMEC co-cultured with T98G (62-63%) within 120-hr incubation with CCSC. Reactive oxygen species (ROS) generated by CCSC in mono-culture of T98G and U-373 MG reached highest levels at 4 and 16 mg/ml, respectively. ROS production by T98G fell when co-cultured with HBMEC or U-373MG. These findings suggest that adverse consequences of CCSC treatment on brain cells may be protected by blood-brain barrier or astrocytes, but with chronic exposure toxicity may be worsened due to destruction of cellular integrity.

Microbial bioconversion and processing methods enhance the phenolic acid and flavonoids and the radical scavenging capacity of Smilax china L. leaf.

BACKGROUND: It has been reported that Smilax china L. leaf (SCL) provided various biological functions owing to polyphenols. The objective of the current study was to assess the enhancing effect of processing methods and microbial conversions on phenolic acid and flavonoid content and radical scavenging capacity of SCL for potential applications of diverse food products. RESULTS: Targeted phenolic acid (chlorogenic acid) and flavonoids (piceid and quercetin) were identified in fresh SCL using liquid chromatography-mass spectrometry. The total amount of identified phenolic acid and flavonoids was highest in steamed SCL (12.70 ± 0.12 mg g(-1) on a dry matter basis, dmb). A substantial amount of chlorogenic acid (5.81 ± 0.16 mg g(-1) dmb), piceid (3.96 ± 0.04 mg g(-1) dmb) and quercetin (6.06 ± 0.12 mg g(-1) dmb) were quantified in SCL fermented by Bacillus species, roasted and steamed, respectively (P < 0.05). The oxygen radical absorbance capacity (ORAC) value was greater in microbial fermented SCL than in others, with the exception of Saccharomyces cerevisiae and Aspergillus oryzae. However, vitamin C equivalent antioxidant capacity (VCEAC) was highest in SCL fermented by Aspergillus oryzae. CONCLUSION: Results from our study suggest that the microbial fermentation processing method could improve accessibility to extraction of phenolic acids and flavonoid content and radical scavenging capacity.

Oxidative Stress Induced by Cigarette Smoke Extracts in Human Brain Cells (T98G) and Human Brain Microvascular Endothelial Cells (HBMEC) in Mono- and Co-Culture.

The objective of the current study was to examine oxidative stress induced by cigarette smoke extract (CSE) or cigarette smoke condensate (CSC) in human brain cells (T98G) and human brain microvascular endothelial cells (HBMEC) in mono- and co-culture systems. Cell viability of T98G cells exposed to CSC (0.05-4 mg/ml) was significantly decreased compared to CSE (0.025-20%). There were no marked differences between quantities of reactive oxygen species (ROS) generation by either CSE (2, 4, and 10%) or CSC (0.2, 0.4, and 0.8 mg/ml) treatment compared to control. However, a significant effect was noted in ROS generation following CSC incubation at 4mg/ml. Cellular integrity of HBMEC decreased to 74 and 64% within 120 h of exposure at the IC50 value of CSE and CSC, respectively. This study suggests that chronic exposure to cigarette smoking might initiate damage to the blood-brain barrier.

Paradigm shift in Nutrient-Energy-Water centered sustainable wastewater treatment system through synergy of bioelectrochemical system and anaerobic digestion.

With advancements in research and the necessity of improving the performance of bioelectrochemical system (BES), coupling anaerobic digestion (AD) with BES is crucial for energy gain from wastewater and bioremediation. Hybridization of BES-AD concept opens new avenues for pollutant degradation, carbon capture and nutrient-resource recovery from wastewater. The strength of merging BES-AD lies in synergy, and this approach was employed to differentiate fads from strategies with the potential for full-scale implementation and making it an energy-positive system. The integration of BES and AD system increases the overall performance and complexity of combined system and the cost of operation. From a technical standpoint, the primary determinants of BES-AD feasibility for field applications are the scalability and economic viability. High potential market for such integrated system attract industrial partners for more industrial trials and investment before commercialization. However, BES-AD with high energy efficacy and negative economics demands performance boost.

Structural basis of IRGB10 oligomerization by GTP hydrolysis.

Immunity-related GTPase B10 (IRGB10) is a crucial member of the interferon (IFN)-inducible GTPases and plays a vital role in host defense mechanisms. Following infection, IRGB10 is induced by IFNs and functions by liberating pathogenic ligands to activate the inflammasome through direct disruption of the pathogen membrane. Despite extensive investigation into the significance of the cell-autonomous immune response, the precise molecular mechanism underlying IRGB10-mediated microbial membrane disruption remains elusive. Herein, we present two structures of different forms of IRGB10, the nucleotide-free and GppNHp-bound forms. Based on these structures, we identified that IRGB10 exists as a monomer in nucleotide-free and GTP binding states. Additionally, we identified that GTP hydrolysis is critical for dimer formation and further oligomerization of IRGB10. Building upon these observations, we propose a mechanistic model to elucidate the working mechanism of IRGB10 during pathogen membrane disruption.

The structure of AcrIC9 revealing the putative inhibitory mechanism of AcrIC9 against the type IC CRISPR-Cas system.

CRISPR-Cas systems are known to be part of the bacterial adaptive immune system that provides resistance against intruders such as viruses, phages and other mobile genetic elements. To combat this bacterial defense mechanism, phages encode inhibitors called Acrs (anti-CRISPR proteins) that can suppress them. AcrIC9 is the most recently identified member of the AcrIC family that inhibits the type IC CRISPR-Cas system. Here, the crystal structure of AcrIC9 from Rhodobacter capsulatus is reported, which comprises a novel fold made of three central antiparallel ß-strands surrounded by three α-helixes, a structure that has not been detected before. It is also shown that AcrIC9 can form a dimer via disulfide bonds generated by the Cys69 residue. Finally, it is revealed that AcrIC9 directly binds to the type IC cascade. Analysis and comparison of its structure with structural homologs indicate that AcrIC9 belongs to DNA-mimic Acrs that directly bind to the cascade complex and hinder the target DNA from binding to the cascade.

Amorphous iron oxide-selenite composite microspheres with a yolk-shell structure as highly efficient anode materials for lithium-ion batteries.

Yolk-shell structured transition metal compounds have intrinsic structural advantages as anode materials and have been synthesized in a highly crystalline form. Thus, development of a synthesis process for a yolk-shell structure with an amorphous state, that displays high structural stability and fast ionic diffusion, is a notable research subject, with wide applications in fields such as energy storage. Herein, a novel approach for synthesizing amorphous materials with a yolk-shell structure using several facile phase transformation processes is presented. Crystalline iron oxide microspheres with a yolk-shell structure were formed by oxidation of the spray-dried product at 400 °C. Using the pitch/tetrahydrofuran solution infiltration method, pitch-infiltrated iron oxide was selenized at 350 °C to form a crystalline iron selenide-C composite. The following partial oxidation process at 375 °C produced a yolk-shell structured amorphous iron oxide-selenite composite. The amorphous characteristics, the yolk-shell structure, and the formation of a heterostructured interface from iron selenite during the initial cycle contributed to high electrochemical kinetic properties and excellent cycling performance of the iron oxide-selenite composite. The amorphous iron oxide-iron selenite yolk-shell microspheres exhibited enhanced reversible capacities, cycling stability, and remarkable electrochemical kinetic properties when compared to crystalline iron oxide.