Hydroxylation with Unusual Stereoinversion Catalyzed by an FeII /2-OG Dependent Oxidase and 3,6-Diene-2,5-diketopiperazine Formation in the Biosynthesis of Brevianamide†K.

Natural products with the 3,6-diene-2,5-diketopiperazine core are widely distributed in nature; however, the biosynthetic mechanism of 3,6-diene-2,5-diketopiperazine in fungi remains to be further elucidated. Through heterologous expression and biochemical investigation of an FeII /2-oxoglutarate-dependent oxidase (AspE) and a heme-dependent P450 enzyme (AspF), we report that AspE, AspF and subsequent dehydration account for the formation of the 3,6-diene-2,5-diketopiperazine substructure of brevianamide K from Aspergillus sp. SK-28, a symbiotic fungus of mangrove plant Kandelia candel. More interestingly, in-depth investigation of the enzymatic mechanism showed that AspE promotes hydroxylation of brevianamide Q with unprecedented stereoinversion through hydrogen atom abstraction and water nucleophilic attack from the opposite face of the resultant iminium cation intermediate.

Buffer capacity regulates the stratification of anode-respiring biofilm during brewery wastewater treatment.

Improving the buffer capacity of the electrolyte can enhance the anode performance in bioelectrochemical systems (BESs). To elucidate the mechanism underlying the facilitated BESs performance, this study used three different anode biofilms cultured with different concentrations of phosphate buffer (5, 50 and 100 mM) to investigate the biofilm response, in terms of the spatial structure of metabolic activity and microbial community, to different buffer capacities. Results showed that the electrochemical activities of the anode biofilms positively correlated with the buffer concentration. The spatial stratification of metabolic activity and microbial community of the anode biofilms were regulated by the buffer capacity, and the spatial microbial heterogeneity of the anode biofilm decreased as the buffer concentration increased. With increasing buffer capacity, Geobacter spp. were enriched in both the inner and outer layers of the biofilm, and the inhibition of methanogens growth improved the COD removal attributed to anode respiration. Additionally, the stimulation of EPS production in biofilms played a role in increasing the electrochemical performance of biofilms by buffer improvement. This study first revealed the regulation of buffer capacity on the stratification of anode biofilm during brewery wastewater treatment, which provided a deep insight into the relation of biofilm structure to its electrochemical properties.

Safety management for polluted confined space with IT system: a running case.

This study traced a deployed real IT system to enhance occupational safety for a polluted confined space. By incorporating wireless technology, it automatically monitors the status of workers on the site and upon detected anomalous events, managers are notified effectively. The system, with a redefined standard operations process, is running well at one of Formosa Petrochemical Corporation's refineries. Evidence shows that after deployment, the system does enhance the safety level by real-time monitoring the workers and by managing well and controlling the anomalies. Therefore, such technical architecture can be applied to similar scenarios for safety enhancement purposes.

Up-regulation of intrarenal renin-agiotensin system contributes to renal damage in high-salt induced hypertension rats.

BACKGROUND/AIMS: To investigate the change of intrarenal renin-agiotensin system (RAS) and its role in high-salt induced hypertension. METHODS: Wistar rats were divided into normal-salt (NS), high-salt diet (HS) and high-salt diet with Losartan group (HS+L), for 6 weeks. Systolic blood pressure (SBP) was monitored. Blood and urine samples were collected every 2 weeks. Angiotensinogen (AGT) was measured by ELISA. AGT mRNA and protein were measured by real-time PCR and immunohistochemistry. Renin activity and angiotensin II (Ang II) were measured by radioimmunoassay. RESULTS: HS versus NS group, SBP increased from 2(nd) week (P<0.05), urinary protein increased at 6(th) week (P<0.05). Although plasma renin, AGT and Ang II had no significant changes (P>0.05), renal cortex renin, AGT, and Ang II increased significantly in HS (P<0.05). In HS+L, Losartan failed to reduce SBP (P>0.05) but abolished the increase of proteinuria (P<0.01), renal cortex renin, AGT, Ang II and urinary AGT reduced (P<0.05) while plasma renin, AGT and Ang II enhanced (P<0.05) when compared with HS. Urinary AGT was positively correlated with renal AGT (r=0.592, P <0.01) and Ang II (r=0.726, P <0.01). CONCLUSION: Inappropriate response of the renal RAS to a high salt diet may contribute to hypertension and renal damage, and urinary AGT could reflect intrarenal RAS activity.

Study of the molecular variation in pre-eclampsia placenta based on micro-Raman spectroscopy.

PURPOSE: Study of the molecular variation in pre-eclampsia placenta based on micro-Raman spectroscopy. METHODS: Five pregnant women with pre-eclampsia from Nanfang hospital were selected as study group whose average age is 28.5 years and 38 ± 2 weeks gestation. The same period of healthy pregnant women, whose average age is 27.6 years and pregnant 39 ± 1 weeks, as control group (n = 5). The normal and pre-eclamptic placental tissues are detected by micro-Raman spectroscopy with the spectrum resolution of 1 cm(-1). RESULTS: We find that the protein structure of α-helix, ß-pleated sheet and ß-turn is overlying in pre-eclamptic placenta, which lead to a disorder of protein structure. The Raman peaks assigned to tryptophan indole ring and phenylalanine in pre-eclamptic placental tissue are more higher than that in normal tissue. CONCLUSIONS: Results suggest that the ordered structures of the main chain in protein molecules are reduced significantly, and the amino acid of side chains is damaged obviously. And a principal component analysis is used to classify the Raman spectra between normal and pre-eclamptic placental tissues. This study presents that Raman spectroscopy has a great potential on the mechanism research and diagnosis of placental lesions.

Observation of Type-II Topological Nodal-Line Fermions in ZrSiSe.

Recently, there has been significant interest in topological nodal-line semimetals due to their linear energy dispersion with one-dimensional nodal lines or loops. These materials exhibit fascinating physical properties, such as drumhead surface states and 3D anisotropic nodal-line structures. Similar to Weyl semimetals, type-II nodal-line semimetals have two crossing bands that are both electron-like or hole-like along a certain direction. However, the direct observation of type-II nodal-line Fermions has been challenging due to the lack of suitable material platforms and the low density of states. Here we present experimental evidence for the coexistence of both type-I and type-II nodal-line Fermions in ZrSiSe, which was obtained through magneto-optical and angle-resolved photoemission spectroscopy (ARPES) measurements. Our density functional theory calculations predict that the type-II nodal-line structure can be developed in the Z-R line of the first Brillouin zone based on the lattice constants of the grown single crystal. Indeed, ARPES measurements reveal the type-II nodal-line band structure. The extracted type-II Landau level transitions from magneto-optical measurements exhibit good agreement with the calculated type-II energy dispersion model based on the band structure. Our experimental results demonstrate that ZrSiSe possesses two types of nodal-line Fermions, distinguishing it from other ZrSiX (X = S, Te) materials and positioning it as an ideal platform for investigating type-II nodal-line semimetals.

Production of the antifungal biopesticide physcion through the combination of microbial fermentation and chemical post-treatment.

Physcion is an anthraquinone compound observed dominantly in medicinal herbs. This anthraquinone possesses a variety of pharmaceutically important activities and has been developed to be a widely used antifungal biopesticide. Herein, we report on the effective preparation of 3R-torosachrysone (4), a tetrahydroanthracene precursor of physcion, in Aspergillus oryzae NSAR1 by heterologous expression of related genes mined from the phlegmacins-producing ascomycete Talaromyces sp. F08Z-0631. Conditions for converting 4 into physcion were studied and optimized, leading to the development of a concise approach for extracting high-purity physcion from the alkali-treated fermentation broth of the 4-producing A. oryzae strain.

[Nonlinear spectral imaging of DNA specimens derived from tumor cells based on second harmonic generation].

Second harmonic generation (SHG) is a second-order nonlinear optical process that has symmetry constraints confining signal to regions lacking a center of symmetry. Using SHG microscopy, a variety of tissue structures have noninvasively been imaged by virtue of intrinsic signal generated by structured proteins such as collagen fibrils in connective tissues or the actomyosin lattice of muscle cells. In biochemistry and structure biology, the high-level structures of DNA and protein macro-molecules are similar in constructing mechanism, although DNAs consist of deoxynucleotides and proteins of amino acid residues. The principal purpose of present work is to detect the SHG signal from different DNA samples by spectral imaging technology based on two-photon excited fluorescence (TPEF) and SHG. These DNA samples include the solution of genomic DNA and extracted nuclei, and cultured living cells. Results show that we can obtain the SHG signal from solution of genomic DNA and extracted nuclei in routine condition, but nothing from cultured cell nuclei. After adding a little of absolute ethanol (less than 5% by volume) in culture medium, the SHG signal is detectable in the interested region of nuclei. The findings suggest that the interaction between ethanol and DNA in living cell gives rise to the shift of molecular conformation, and this shift changes some nonlinear optical properties of DNA molecules.

[Raman spectroscopic study on the influence of ultraviolet-A radiation on collagen I].

Raman spectroscopy was used to study the influence of ultraviolet-A(UV-A) radiation on collagen I. The Raman spectra of collagen I and that after 90 min UV-A radiation were reported. The results proved that irradiation with 90 min UV-A caused the change in the structures of collagen I. Intramolecular hydrogen bonds were broken, and the hydrogen bonding system was changed. The intensity of helix was decreased, while the intensity of the disordered conformation in proteins such as random coil was increased. Otherwise, the UV-A radiation influenced the hydroxylation of proline and the content of hydroxyproline was reduced. The changes caused by UV-A radiation could damage the triple helical structure of collagen I. It would lead to a series of changes, such as the destruction of collagen fibers during the photoaging of skin.

The Role of Exopolysaccharides in Direct Interspecies Electron Transfer.

Direct interspecies electron transfer (DIET) is an effective mechanism for microbial species to exchange electrons cooperatively during syntrophic metabolism. It is generally accepted that DIET is mainly mediated by electrically conductive pili and outer surface c-type cytochromes (c-Cyts). However, as an extracellular matrix is ubiquitous and abundant on the surface of microorganisms, the effect and mechanism of exopolysaccharides on DIET are still unclear. This study constructed a co-culture of exopolysaccharides-deficient Geobacter sulfurreducens with Geobacter metallireducens to explore the role of exopolysaccharides in DIET. Results revealed that the deficiency of exopolysaccharides extended the metabolic period of the co-culture by 44.4% and changed the proportions of each species in the co-culture. The exopolysaccharides-deficient co-culture failed to form large, tight spherical aggregates and the expression of c-Cyts and pili was decreased. The addition of magnetite and granular activated carbon (GAC), respectively, might compensate for the functions of c-Cyts and pili in the first generation of co-culture, but the stimulatory effect on the metabolic stable period co-culture was fairly limited. These findings demonstrate that non-conductive exopolysaccharides are an important component of DIET aggregates and an extracellular matrix for DIET-required c-Cyts.

Exopolysaccharides matrix affects the process of extracellular electron transfer in electroactive biofilm.

The applications of bioelectrochemical systems (BESs) in the field of environment and energy are achieved through the bioelectrocatalytic process of electroactive biofilms. As a primary component of biofilm, the role of exopolysaccharides in electroactive biofilm in BESs is poorly understood. This study constructed an exopolysaccharides-deficient Geobacter sulfurreducens-based BES to explore the role of exopolysaccharides in electroactive biofilm. Compared with the wild type, the mutant biofilm expressing less exopolysaccharides decreased the capacity of current generation. In the mutant biofilm, the content of exopolysaccharides decreased significantly, resulting in a thinner biofilm and lower cell viability compared with the wild-type biofilm. However, the mutant with overexpressed pili developed a mature biofilm with extended time, which indicating the importance of exopolysaccharides for early biofilm formation and the compensatory role of pili in biofilm formation. The mutant biofilm had less content of c-type cytochromes (c-Cyts) and lower electrochemical activity of extracellular polymeric substances than the wild-type biofilm, suggesting a function of exopolysaccharides anchoring extracellular c-Cyts that essential to extracellular electron transfer (EET) in electroactive biofilms. Our findings demonstrated the essential role of exopolysaccharides in the process of EET in electroactive biofilm, which contributed to a better understanding and optimization of the performance of BESs.

Unsymmetrically Regioselective Homodimerization Depends on the Subcellular Colocalization of Laccase/Fasciclin Protein in the Biosynthesis of Phlegmacins.

Phlegmacins are homodimeric dihydroanthracenone natural products featuring two torosachrysone monomers unsymmetrically conjugated by 7,10'-coupling. Herein, we report the identification and characterization of the biosynthetic gene cluster of phlegmacins in ascomycete Talaromyces sp. F08Z-0631. On the basis of the heterologous reconstitution of the phlegmacin pathway in Aspergillus oryzae, we demonstrated an unprecedented laccase-involved unsymmetrically regioselective oxidative coupling reaction. The association of laccase PhlC and the fasciclin partner protein PhlB was verified to be indispensable for the coupling activity. Intriguingly, both proteins can be transferred and located independently at the mitochondrial membrane. Notably, only their subcellular colocalization led to the occurrence of oxidative dimerization. These observations add new insights into the poorly understood catalytic mechanisms of various laccases involved in the biosynthesis of secondary metabolites, particularly those functioning with variable partners.

A Comprehensive Study of the Mechanical and Durability Properties of High-Performance Concrete Materials for Grouting Underwater Foundations of Offshore Wind Turbines.

With the increasing importance of offshore wind turbines, a critical issue in their construction is the high-performance concrete (HPC) used for grouting underwater foundations, as such materials must be better able to withstand the extremes of the surrounding natural environment. This study produced and tested 12 concrete sample types by varying the water/binder ratio (0.28 and 0.30), the replacement ratios for fly ash (0%, 10%, and 20%) and silica fume (0% and 10%), as substitutes for cement, with ground granulated blast-furnace slag at a fixed proportion of 30%. The workability of fresh HPC is discussed with setting time, slump, and V-funnel flow properties. The hardened mechanical properties of the samples were tested at 1, 7, 28, 56, and 91 days, and durability tests were performed at 28, 56, and 91 days. Our results show that both fly ash (at 20%) and silica fume (at 10%) are required for effective filling of interstices and better pozzolanic reactions over time to produce HPC that is durable enough to withstand acid sulfate and chloride ion attacks, and we recommend this admixture for the best proportioning of HPC suitable for constructing offshore wind turbine foundations under the harsh underwater conditions of the Taiwan Bank. We established a model to predict a durability parameter (i.e., chloride permeability) of a sample using another mechanical property (i.e., compressive strength), or vice versa, using the observable relationship between them. This concept can be generalized to other pairs of parameters and across different parametric categories, and the regression model will make future experiments less laborious and time-consuming.

Redox potential-induced regulation of extracellular polymeric substances in an electroactive mixed community biofilm.

Electroactive biofilms are promising in achieving efficient wastewater treatment and energy conversion in bioelectrochemical systems (BESs). Extracellular polymeric substances (EPS) are important for physical contact with electrode surface and extracellular electron transfer (EET) within biofilm. Redox potential is an important trigger for the regulation of EPS in microbial aggregates, but this yet is lacking for electroactive mixed community biofilms. This study first explored how redox potential affected EPS of electroactive mixed community biofilms, which were cultured in BES reactors with different anode potentials (-0.3 V, 0 V, +0.3 V, +0.6 V vs. SCE) using artificial brewery wastewater as substrate. The anode potential regulated biocurrent generation, overall EPS production, EPS composition and EPS redox properties. The biofilms poised at 0 V exhibited the highest current production (7.2 mA) and EPS redox capacity, while the +0.6 V biofilms had the lowest current production (1.2 mA) with lowest EPS redox capacity. The steady-state current exhibited a significant positive correlation with EPS redox capability, suggesting an important role of EPS in anode potential-dependent current production. Significant positive correlations between proteins or humic substances in EPS and EPS redox properties further verified that EPS redox feature raised from proteins and humic substances. This study provided a potential mechanism that anode potential determined the electroactivity of anode biofilm via regulating EPS composition and redox properties, and will facilitate the use of electroactive biofilms in bioelectrochemical applications.

Co-selection of antibiotic-resistant bacteria in a paddy soil exposed to As(III) contamination with an emphasis on potential pathogens.

The increased acquisition of antibiotic resistance by pathogens is a global health concern. The environmental selection of antibiotic resistance can be caused by either antibiotic residues or co-selecting agents such as toxic metal(loid)s. This study explored the potential role of As(III) as a co-selecting driver in the spread of antibiotic resistance in paddy soils. By applying high-throughput sequencing, we found that the diversity and composition of soil microbial communities was significantly altered by As(III) exposure, resulting in an increased proportion of potential pathogens (9.9%) compared to the control soil (0.1%). Meanwhile, a total of 46 As(III)-resistant isolates were obtained from As(III)-exposure soil, among which potential pathogens accounted for 54.3%. These As(III)-resistant bacteria showed a high incidence of resistance to sulfanilamide (100%) and streptomycin (88-93%). The association between antibiotic and As(III) resistances was further investigated in a potentially pathogenic isolate by whole-genome sequencing and a transcription assay. The results showed that As(III) and antibiotic resistance genes might co-occur in a mobile genomic island and be co-regulated by As(III), implying that antibiotic resistance could be co-selected by As(III) via co-resistance and co-regulation mechanisms. Overall, these results suggest that As(III) exposure provides a strong selective pressure for the expansion of soil bacterial resistome.

miR-4295 promotes cell proliferation, migration and invasion of osteosarcoma through targeting interferon regulatory factor 1.

Osteosarcoma (OS) is the most common form of primary malignant bone tumor. Despite encouraging progress in the treatment of OS, the survival rate for patients with OS has remained unchanged over the past 40 years. It has been established that miRNA plays a crucial regulatory role in the progression and development of OS. To explore the potential association of miRNAs with OS, bioinformatics techniques were used to screen for differentially expressed miRNA genes in OS in the Gene Expression Omnibus database. In the GSE70367 database, it was revealed that miR-4295 expression was abnormally elevated in the expression of OS cells. To characterize the potential function of miR-4295 in OS, the expression levels of miR-4295 in 30 samples of OS and adjacent normal tissues was examined. The results revealed that the expression of miR-4295 was significantly increased in OS tissues compared with the paired normal tissues. Moreover, the expression levels of miR-4295 in OS cell lines (MG-63 and Saos-2) were significantly higher compared with those in the normal human mesenchymal stem cells. In addition, miR-4295 was associated with OS cell proliferation, migration and invasion. Furthermore, it was demonstrated that the expression of interferon regulatory factor (IRF)1, a tumor suppressor, was regulated by miR-4295 directly in OS cells. Taken together, the present results revealed that miR-4295 may act as a tumor activator by targeting IRF1 during the progression of OS. Investigating miR-4295 may provide novel insight into the mechanisms of OS metastasis, and inhibition and targeting miR-4295 may be a novel therapeutic strategy for the treatment of OS.

Physiological potential of extracellular polysaccharide in promoting Geobacter biofilm formation and extracellular electron transfer.

Geobacter sulfurreducens biofilms have promising applications in renewable energy, pollutant bioremediation, and bioelectronic applications. Genetically manipulating G. sulfurreducens biofilms is an effective strategy to improve the capacity of extracellular electron transfer (EET). Extracellular polysaccharide, a sticky component surrounding microbes, plays an important role in EET. Herein, we constructed a mutant of G. sulfurreducens strain PCA overexpressing the gene GSU1501 (part of the ATP-dependent exporter of the polysaccharide biosynthesis gene operon), designated strain PCA-1501, to increase EET capacity. Experimental results showed that the overexpression of GSU1501 increased extracellular polysaccharide secretion by 25.5%, which promoted the formation of biofilm with higher thickness and viability, as well as the content of extracellular c-type cytochromes. Compared with the control strain, the mutant showed a higher capacity of Fe(III) oxide reduction and current generation (increased by 20.4% and 22.2%, respectively). Interestingly, the overexpression of GSU1501 hindered the pili formation by reducing the transcription level of pilA; a compensatory relationship between extracellular polysaccharide and pili in promoting biofilm formation deserves further investigation. This study provides a feasible method to promote the EET capacity of G. sulfurreducens biofilms, which benefit their bioelectrochemical applications.

A novel tri-band T-junction impedance-transforming power divider with independent power division ratios.

In this paper, a novel L network (LN) is presented, which is composed of a frequency-selected section (FSS) and a middle stub (MS). Based on the proposed LN, a tri-band T-junction power divider (TTPD) with impedance transformation and independent power division ratios is designed. Moreover, the closed-form design theory of the TTPD is derived based on the transmission line theory and circuit theory. Finally, a microstrip prototype of the TTPD is simulated, fabricated, and measured. The design is for three arbitrarily chosen frequencies, 1 GHz, 1.6 GHz, and 2.35 GHz with the independent power division ratios of 0.5, 0.7, and 0.9. The measured results show that the fabricated prototype is consistent with the simulation, which demonstrates the effectiveness of this proposed design.

Altered peripheral profile of blood cells in Alzheimer disease: A hospital-based case-control study.

Alzheimer disease (AD) has been made a global priority for its multifactorial pathogenesis and lack of disease-modifying therapies. We sought to investigate the changes of profile of blood routine in AD and its correlation with the disease severity.In all, 92 AD patients and 84 age and sex-matched normal controls were enrolled and their profiles of blood routine were evaluated.Alzheimer disease patients had increased levels of mean corpuscular hemoglobin, mean corpuscular volume, red cell distribution width-standard deviation, mean platelet volume,and decreased levels of platelet distribution width, red blood cell, hematocrit, hemoglobin, lymphocyte, and basophil compared with normal controls.Alterations in quantity and quality of blood cells may be involved in the pathogenesis of AD and contribute to the disease progression.

SMPD1 variants in Chinese Han patients with sporadic Parkinson's disease.

INTRODUCTION: A founder mutation, p.L302P, in sphingomyelin phosphodiesterase 1, acid lysosomal (SMPD1), causing Niemann-Pick disease, a recessive lysosomal storage disorder, was reported to be associated with increased risk of Parkinson's disease (PD) in Ashkenazi Jewish population. Several other studies about the association between SMPD1 variants and PD were performed afterward in other populations. However, the results on the role of SMPD1 mutations for PD have been conflicting. This study aimed to investigate the role of mutations in SMPD1 in Chinese PD patients. METHODS: We sequenced all the exons of this gene in 512 Chinese Han cases with sporadic Parkinson's disease and 495 matched healthy control subjects. RESULTS: We identified Leu-Ala (Val) repeat variants and six known single nucleotide variants (p.A36V, p.D212D, p.P332R, p.G508R, p.P533L, p.T544T) in SMPD1 in both patients and normal controls. Case-control analysis showed the association between Leu-Ala (Val) repeat variants in SMPD1and Chinese Han patients with PD (χ2 = 8.771, p = 0.012), and the allele with less than seven LeuAla (Val) repeats may increase the risk of PD (p = 0.010). CONCLUSION: We identified association between Leu-Ala (Val) repeat variants in SMPD1 and Chinese Han patients with sporadic Parkinson's disease. Our results provide further support for the role of lysosomal pathways in PD development.