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Through iterative rounds of mutation and selection, proteins can be engineered to enhance their desired biological functions. Nevertheless, identifying optimal mutation sites for directed evolution remains challenging due to the vastness of the protein sequence landscape and the epistatic mutational effects across residues. To address this challenge, we introduce MLSmut, a deep learning-based approach that leverages multi-level structural features of proteins. MLSmut extracts salient information from protein co-evolution, sequence semantics, and geometric features to predict the mutational effect. Extensive benchmark evaluations on 10 single-site and two multi-site deep mutation scanning datasets demonstrate that MLSmut surpasses existing methods in predicting mutational outcomes. To overcome the limited training data availability, we employ a two-stage training strategy: initial coarse-tuning on a large corpus of unlabeled protein data followed by fine-tuning on a curated dataset of 40-100 experimental measurements. This approach enables our model to achieve satisfactory performance on downstream protein prediction tasks. Importantly, our model holds the potential to predict the mutational effects of any protein sequence. Collectively, these findings suggest that our approach can substantially reduce the reliance on laborious wet lab experiments and deepen our understanding of the intricate relationships between mutations and protein function., (© 2024 Wiley‐VCH GmbH.)

The central nervous system HIV reservoir is incompletely understood and is a major barrier to HIV cure. We profiled people with HIV (PWH) and uninfected controls through single-cell transcriptomic and T cell receptor (TCR) sequencing to understand the dynamics of HIV persistence in the CNS. In PWH on ART, we found that most participants had single cells containing HIV-1 RNA, which was found predominantly in CD4 central memory T cells, in both cerebrospinal fluid (CSF) and blood. HIV-1 RNA-containing cells were found more frequently in CSF than blood, indicating a higher burden of reservoir cells in the CNS than blood for some PWH. Most CD4 T cell clones containing infected cells were compartment specific, while some (22%) - including rare clones with members of the clone containing detectable HIV RNA in both blood and CSF - were found in both CSF and blood. These results suggest that infected T cells trafficked between tissue compartments and that maintenance and expansion of infected T cell clones contributed to the CNS reservoir in PWH on ART.

Pseudomonas putida KT2440, a GRAS strain, has been used for synthesizing bulk and fine chemicals. However, the gene editing tool to metabolically engineer KT2440 showed low efficiency. In this study, a novel sacB-based system pK51mobsacB was established to improve the efficiency for marker-free gene disruption. Then the rhamnolipid synthetic pathway was introduced in KT2440 and genes of the competitive pathways were deleted to lower the metabolic burden based on pK51mobsacB. A series of endogenous and synthetic promoters were used for fine tuning rhlAB expression. The limited supply of dTDP-L-rhamnose was enhanced by heterologous rmlBDAC expression. Cell growth and rhamnolipid production were well balanced by using glucose/glycerol as mixed carbon sources. The final strain produced 3.64 g/L at shake-flask and 19.77 g/L rhamnolipid in a 5 L fermenter, the highest obtained among metabolically engineered KT2440, which implied the potential of KT2440 as a promising microbial cell factory for industrial rhamnolipid production., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

IL-15 is under clinical investigation toward the goal of curing HIV infection because of its abilities to reverse HIV latency and enhance immune effector function. However, increased potency through combination with other agents may be needed. 3-Hydroxy-1,2,3-benzotriazin-4(3H)-one (HODHBt) enhances IL-15-mediated latency reversal and NK cell function by increasing STAT5 activation. We hypothesized that HODHBt would also synergize with IL-15, via STAT5, to directly enhance HIV-specific cytotoxic T cell responses. We showed that ex vivo IL-15 + HODHBt treatment markedly enhanced HIV-specific granzyme B-releasing T cell responses in PBMCs from antiretroviral therapy-suppressed (ART-suppressed) donors. We also observed upregulation of antigen processing and presentation in CD4+ T cells and increased surface MHC-I. In ex vivo PBMCs, IL-15 + HODHBt was sufficient to reduce intact proviruses in 1 of 3 ART-suppressed donors. Our findings reveal the potential for second-generation IL-15 studies incorporating HODHBt-like therapeutics. Iterative studies layering on additional latency reversal or other agents are needed to achieve consistent ex vivo reservoir reductions.

Transgender women (TGW) are disproportionally affected by HIV infection, with a global estimated prevalence of 19.9%, often attributed to behavioral risk factors, with less known about biological factors. We evaluated potential biological risk factors for HIV acquisition in TGW at the sites of viral entry by assessing immune parameters of the neovaginal surface and gut mucosa. The neovagina in TGW, compared with the vagina in cisgender women (CW), shows distinct cell composition and may pose a more inflammatory environment, evidenced by increased CD4+ T cell activation and higher levels of soluble markers of inflammation (C-reactive protein, soluble CD30). Increased inflammation may be driven by microbiome composition, as shown by a greater abundance of Prevotella and a higher Shannon Diversity Index. In addition, we have observed higher frequency of CD4+CCR5+ target cells and decreased DNA methylation of the CCR5 gene in the gut mucosa of TGW compared with CW and men who have sex with men, which was inversely correlated with testosterone levels. The rectal microbiome composition in TGW appears to favor a proinflammatory milieu as well as mucosal barrier disruption. Thus, it is possible that increased inflammation and higher frequencies of CCR5-expressing target cells at sites of mucosal viral entry may contribute to increased risk of HIV acquisition in TGW, with further validation in larger studies warranted.

Sugar transporters play an important role in the cellulase production of lignocellulose-degrading fungi. Nevertheless, the role and function of these transporters are still unclear. Here we first report intracellular sugar transporters assisting cellulase production in Trichoderma reesei ( T. reesei ) using lactose. The mRNA levels of sugar transporter genes mfs , gst , and lac1 were substantially upregulated in T. reesei cultivated on lactose, with the most abundant mRNA levels at 24 h as compared to glucose. Moreover, the individual deletion of these sugar transporters significantly inhibited cellulase production, solid cell growth, and sporulation of T. reesei , suggesting they play a supporting role in cellulase production when grown in lactose. Surprisingly, MFS, GST, and LAC1 were mainly localized in the cytoplasm, with MFS and LAC1 in the endoplasmic reticulum (ER), representing the first discovery of intracellular sugar transporters involved in cellulase biosynthesis in lactose culture. The absence of the gene lac1 noticeably inhibited most of the crucial genes related to cellulase production, including cellulase-encoding genes, transcription factors, and sugar transporters, at 24 h, which was fully relieved at 48 h or 72 h, indicating that lac1 affects cellulase production more at the early step. This research advances the understanding of the function of intracellular sugar transporters in fungi, particularly for fungal cellulase production.

Background: In the process of cellulose hydrolysis, carbohydrate hydrolysates are transported into cells through membrane transporters, and then affect the expression of cellulase-encoding genes. Sugar transporters play a crucial role in cellulase production in lignocellulolytic fungi, of which relatively few have been functionally validated to date and are all reported to be on cell membrane., Result: Through transcriptome analysis and qRT-PCR, three putative MFS sugar transporters GST, MFS, and LAC1 were found to display significantly higher mRNA levels in T. reesei grown on cellulose than on glucose. The individual deletion of these three genes compromised cellulase production and delayed sugar absorption by 24 h in T. reesei. Nevertheless, they transported pretty low level of sugars, including galactose, lactose, and mannose, and did not transport glucose, when expressed in yeast system. Meanwhile, all three transporters were unexpectedly found to be intracellular, being located in endoplasmic reticulum (ER). Particularly, the knockout of lac1 almost abolished cellulase production, and significantly inhibited biomass generation regardless of sugar types, indicating that lac1 is essential for cellulase production and biomass formation. The absence of lac1 upregulated genes involved in ribosome biogenesis, while downregulated genes in cellulase production, protein processing in ER (particularly protein glycosylation), and lipid biosynthesis. The inhibition of lac1 deletion on the transcriptional levels of genes related to cellulase biosynthesis was restored after 72 h, but the cellulase production was still inhibited, indicating lac1 might pose a post-transcription regulation on cellulase production that are independent on the known cellulase regulation mediated by CRT1 and XYR1., Conclusion: For the first time, intracellular sugar transporters (mfs, gst, and lac1) facilitating cellulase production were identified, which was distributed in ER. Their sugar transporting ability was very weak, indicating that they might be related to sugar utilization inside cells rather than the cellular sugar uptake. More importantly, sugar transporter lac1 is first found to be essential for cellulase production and biomass formation by affecting protein processing in ER (particularly protein glycosylation) and lipid biosynthesis. The effect of LAC1 on cellulase production seems to be post-transcriptional at late stage of cellulase production, independent on the well-known cellulase regulation mediated by CRT1 and XYR1. These findings improve the understanding of intracellular sugar transporters in fungi and their important role in cellulase synthesis., (© 2022. The Author(s).)

Given the significant impact of ions on environment pollution and human health, it is urgently needed to establish effective and convenient ion detection approaches, particularly in living cells. In this paper, we constructed multicolor N-doped-carbon dots (mPD-CDs) by facile one-step hydrothermal carbonization of m-phenylenediamine (mPD). mPD-CDs were successfully deployed for multicolor cellular imaging for animal cells, fungi, and bacteria in a wash-free way with high photostability and satisfactory biocompability. Moreover, mPD-CDs can be used as a fluorescent sensing probe for ultrasensitive detection of both iodide ion (I - ) and typical heavy metals such as cadmium (Cd 2+ ), copper (Cu 2+ ), mercury (Hg 2+ ), gadolinium (Gd 3+ ), ferrous ion (Fe 2+ ), Zinc (Zn 2+ ), and ferric ion (Fe 3+ ). This is the first report using CDs as optical sensing probe for the detection of Gd 3+ , and for detection of Fe 3+ with fluorescence "turn on". More significantly, with these versatile and fascinating properties, we applied mPD-CDs for intracellular ion detection in living cells like Hep G2 and S. cerevisiae, and zebra fish. Altogether, mPD-CDs displayed great potential for multicolor cell imaging and the multiple ion detection in vitro and in vivo, presenting a promising strategy for in-situ ultrasensitive sensing of multiple metal ions in the environment and the biological systems., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Background: A total of 11 β-glucosidases are predicted in the genome of Trichoderma reesei, which are of great importance for regulating cellulase biosynthesis. Nevertheless, the relevant function and regulation mechanism of each β-glucosidase remained unknown., Results: We evidenced that overexpression of cel1b dramatically decreased cellulase synthesis in T. reesei RUT-C30 both at the protein level and the mRNA level. In contrast, the deletion of cel1b did not noticeably affect cellulase production. Protein CEL1B was identified to be intracellular, being located in vacuole and cell membrane. The overexpression of cel1b reduced the intracellular pNPGase activity and intracellular/extracellular glucose concentration without inducing carbon catabolite repression. On the other hand, RNA-sequencing analysis showed the transmembrane transport process and endoplasmic reticulum function were affected noticeably by overexpressing cel1b. In particular, some important sugar transporters were notably downregulated, leading to a compromised cellular uptake of sugars including glucose and cellobiose., Conclusions: Our data suggests that the cellulase inhibition by cel1b overexpression was not due to the β-glucosidase activity, but probably the dysfunction of the cellular transport process (particularly sugar transport) and endoplasmic reticulum (ER). These findings advance the knowledge of regulation mechanism of cellulase synthesis in filamentous fungi, which is the basis for rationally engineering T. reesei strains to improve cellulase production in industry., (© 2022. The Author(s).)

Background: Knowledge on regulatory networks associated with cellulase biosynthesis is prerequisite for exploitation of such regulatory systems in enhancing cellulase production with low cost. The biological functions of intron retention (IR) and nonsense-mediated mRNA decay (NMD) in filamentous fungi is lack of study, let alone their roles in cellulase biosynthesis., Results: We found that major cellulase genes (cel7a, cel7b, and cel3a) exhibited concomitant decrease in IR rates and increase in their gene expression in T. reesei under cellulase-producing condition (cellulose and lactose) that was accompanied with a more active NMD pathway, as compared to cellulase non-producing condition (glucose). In the presence of the NMD pathway inhibitor that successfully repressed the NMD pathway, the mRNA levels of cellulase genes were sharply down-regulated, but the rates of IR in these genes were significantly up-regulated. Consistently, the cellulase activities were severely inhibited. In addition, the NMD pathway inhibitor caused the downregulated mRNA levels of two important genes of the target of rapamycin (TOR) pathway, trfkbp12 and trTOR1. The absence of gene trfkbp12 made the cellulase production in T. reesei more sensitive to the NMD pathway inhibitor., Conclusions: All these findings suggest that the IR of cellulase genes regulates their own gene expression by coupling with the NMD pathway, which might involve the TOR pathway. Our results provide better understanding on intron retention, the NMD pathway, and cellulase production mechanism in filamentous fungi., (© 2022. The Author(s).)

To reduce the bleeding time and to shorten the surgery time are vital to patients' prog-nosis, therefore, in this study, high moisture absorption nonwoven composites are proposed to attain hemostasis in time. Polyacrylate fiber and Tencel ® fibers at different blending ratios (10:90, 20:80, 30:70, 40:60, and 50:50) are used to form PT composite nonwoven. Next, composed of a 50:50 ratio, PT composite nonwoven exhibits the maximal vertical wicking height of 4.4 cm along the cross direction. Additionally, the UV-Vis absorption spectra analysis shows that at absorption waves of 413-415 nm, the occurring of distinct peaks suggests the presence of nanoparticles. The XRD patterns indicate the presence of silver nanoparticles with corresponding crystal planes of characteristic peaks at (111), (200), and (220). Polyacrylate/Tencel ® nonwoven composites exhibit comparable adsorption capacity of blood and water molecules. In particular, 30PT composite nonwoven outperforms the control group, exhibiting 3.8 times and 4.7 times greater the water absorption and blood absorption, respectively. Moreover, a great number of red blood cells with a size of 4-6 μm agglomerate among fibers as observed in SEM images, while 6hr-PT composite dressing demonstrates the optimal antibacterial efficacy against Escherichia coli and Staphylococcus aureus, proven by the zone of inhibition being 1.9 mm and 0.8 mm separately. When in contact with plasma, hemostasis composites have plasma hemostasis prothrombin time of 97.9%, and activated partial thromboplastin time of 96.7%. As for animal hemostasis model, the arteria over the rats' thigh bones is cut open perpendicularly, generating mass arteria hemorrhage. To attain hemostasis, it takes 46.5% shorter time when using composite dressings (experimental group) than the control group.

The development of micro/nanofluidic techniques has recently revived interest in dynamic shear flow at liquid-solid interfaces. When the nature of the liquid-solid boundaries was revisited, the slip of the fluids relative to the solid wall was predicted theoretically and confirmed experimentally. This indicates that the molecular-level structures of the liquid-solid interfaces will be influenced by the liquid flow over certain temporal and spatial criteria. However, the fluid flow at the boundary layer still cannot be precisely predicted and effectively controlled, somehow limiting its practical applications. Here, we summarize the recent advances for the microscopic structures at the liquid-solid interfaces upon shear flow. Special attention was given to a second-order nonlinear optical technique, sum frequency generation vibrational spectroscopy, which is a powerful tool for exploring the molecular-level structures and structural dynamics at the liquid-solid interfaces and offering new insights into the molecular mechanisms of the fluid slip at the interfaces. Moreover, we discuss the possible approaches for controlling the interfacial slip at the molecular level and highlight the current challenges and opportunities. Although the theoretical framework of the slip at the liquid-solid interfaces is still incomplete, we hope that this Perspective will complement and enhance our understanding of various interfacial properties and phenomena with respect to practical non-equilibrium dynamic processes happening at the interfaces.

Congo red (CR) is a hazardous pigment, posing increasing dangerous to the environment and human health. However, the in-situ detection of CR in living cells has not been reported, as far as we know. Here, negatively-charged green-emitting Ca, N, S-doped carbon dots (Mis-mPD-CDs) were fabricated from plant and m-phenylenediamine (mPD) by facile one-step hydrothermal carbonization. Mis-mPD-CDs were capable of rapidly detecting CR on the basis of their fluorescence quenching by CR due to the inner filter effect. This CR detection based on Mis-mPD-CDs displayed a linear range of 0.2-1.2 μM and a low limit of detection (58 nM), and was not interfered by metal ions, important biological molecules, and other dyes, showing high sensitivity and selectivity. More interestingly, Mis-mPD-CDs can rapidly enter and label animal cells (A549, 4T1, and HUVEC), fungi (S. cerevisiae, C. albicans, and T. reesei), and bacteria (E. coli and S. aureus) for long term with high stability and appealing biocompatibility. Based on these compelling characteristics, we applied Mis-mPD-CDs for sensing and imaging CR in living cells (A549, C. albicans, E. coli, and S. aureus) and zebra fish. On the other hand, the quantitative detection of CR by Mis-mPD-CDs was realized in real samples like fish tissues and industrial wastewater. This is the first report on applying CDs for rapid CR detection in living cells and in vivo. Mis-mPD-CDs provides a novel efficient platform for probing intracellular CR, expanding the applications of CDs as biosensors for toxic dyes., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

The measurement of fungal cell growth in submerged culture systems containing insoluble compounds is essential yet difficult due to the interferences from the insoluble compounds like biopolymers. Here, we developed a fluorescent strategy based on chitosan-modified fluorescein isothiocyanate (GC-FITC) to monitor the cell growth of lignocellulosic fungi cultivated on biopolymers. GC-FITC could stain only lignocellulosic fungi (Tricoderma reesei, Penicillium oxalicum, Aspergillus nidulans, and Neurospora crassa), but not biopolymers (cellulose, xylan, pectin, or lignin), excluding the interferences from these insoluble biopolymer. Moreover, a linear relationship was observed between the fluorescence intensity of GC-FITC absorbed by lignocellulosic fungi and the biomass of lignocellulosic fungi. Therefore, GC-FITC was leveraged to monitor the cell growth of lignocellulosic fungi when using biopolymers like cellulose as the carbon sources, which is faster, more convenient, time-saving, and cost-effective than the existing methods using protein/DNA content measurement. GC-FITC offers a powerful tool to detect fungal growth in culture systems with insoluble materials., (© The Author(s) 2022. Published by Oxford University Press on behalf of FEMS.)

Revealing interfacial shear-induced structural responsiveness has long been an important topic in that most fluids in nature and human life are in motion and cause interesting boundary phenomena. It is amazing how the polymer chain conformation or local structural features at a boundary change under the effective shear condition. In this study, microfluidic-assisted sum frequency generation (SFG) vibrational spectroscopy and all-atom molecular dynamics (MD) simulation are combined to reveal that the shear flow can effectively block the so-called thermal coil-to-globule transition of the poly( N -isopropylacrylamide) (PNIPAM) brushes on the solid substrate, and the normal coil-to-globule transition transfers to a coil-to-stretch one under shear flow with increasing ambient temperature. Such findings are attributed to the balance between the shear flow and the molecular interaction with respect to the polymer chains and adjacent water molecules, thus demonstrating the significant effect of the shear flow on the structural and dynamic behaviors of the polymer chains at the boundaries from the molecular level.

Phase diagram of Au-Si binary alloy system shows a large drop in melting temperature of about 1000K compared with that of Si at a composition of Au:Si = 81:19, where the melting temperature is about 636K. Mixing of Au and Si below the melting temperature was observed by transmission electron microscopy experiment, and it was found that the mixed region shows a diffraction pattern of a diffuse ring intensity indicating an amorphous structure of the mixed area. Si L-emission spectra, which reflect the energy state of bonding electrons of Si atom, of molten Au81Si19 alloy were measured for the first time to investigate the energy state of valence electrons of Si. The Si L-emission spectrum showed a characteristic loss of L1 peak, which is related to sp3 directional bonding in crystalline Si. The intensity profile is also different from that of molten Si reported. This suggests a characteristic atomic arrangement that exists in the molten state. The intensity profile also indicated a small density of state in the molten state at Fermi energy. The obtained spectrum was compared with the calculated density of state of possible crystal structures reported. The comparison suggested that Si atoms are surrounded by eight Au atoms in the molten state of Au81Si19 alloy. The formation of this local atomic arrangement can be an origin of a large drop of melting temperature at about Au:Si = 81:19., (© The Author(s) 2021. Published by Oxford University Press on behalf of The Japanese Society of Microscopy. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Background: Cellulase synthesized by fungi can environment-friendly and sustainably degrades cellulose to fermentable sugars for producing cellulosic biofuels, biobased medicine and fine chemicals. Great efforts have been made to study the regulation mechanism of cellulase biosynthesis in fungi with the focus on the carbon sources, while little attention has been paid to the impact and regulation mechanism of nitrogen sources on cellulase production., Results: Glutamine displayed the strongest inhibition effect on cellulase biosynthesis in Trichoderma reesei, followed by yeast extract, urea, tryptone, ammonium sulfate and L-glutamate. Cellulase production, cell growth and sporulation in T. reesei RUT-C30 grown on cellulose were all inhibited with the addition of glutamine (a preferred nitrogen source) with no change for mycelium morphology. This inhibition effect was attributed to both L-glutamine itself and the nitrogen excess induced by its presence. In agreement with the reduced cellulase production, the mRNA levels of 44 genes related to the cellulase production were decreased severely in the presence of glutamine. The transcriptional levels of genes involved in other nitrogen transport, ribosomal biogenesis and glutamine biosynthesis were decreased notably by glutamine, while the expression of genes relevant to glutamate biosynthesis, amino acid catabolism, and glutamine catabolism were increased noticeably. Moreover, the transcriptional level of cellulose signaling related proteins ooc1 and ooc2, and the cellular receptor of rapamycin trFKBP12 was increased remarkably, whose deletion exacerbated the cellulase depression influence of glutamine., Conclusion: Glutamine may well be the metabolite effector in nitrogen repression of cellulase synthesis, like the role of glucose plays in carbon catabolite repression. Glutamine under excess nitrogen condition repressed cellulase biosynthesis significantly as well as cell growth and sporulation in T. reesei RUT-C30. More importantly, the presence of glutamine notably impacted the transport and metabolism of nitrogen. Genes ooc1, ooc2, and trFKBP12 are associated with the cellulase repression impact of glutamine. These findings advance our understanding of nitrogen regulation of cellulase production in filamentous fungi, which would aid in the rational design of strains and fermentation strategies for cellulase production in industry., (© 2021. The Author(s).)

Icosahedral Al-Cu-Fe quasicrystal (QC) shows moderate electrical conductivity and low thermal conductivity, and both p- and n-type conduction can be controlled by tuning the sample composition, making it potentially suited for thermoelectric materials. In this work, we investigated the effect of introducing chemical disorder through heavy element substitution on the thermal conductivity of Al-Cu-Fe QC. We substituted Au and Pt elements for Cu up to 3 at% in a composition of Al 63 Cu 25 Fe 12 , i.e., Al 63 Cu 25- x (Au,Pt) x Fe 12 ( x = 0, 1, 2, 3). The substitutions of Au and Pt for Cu reduced the phonon thermal conductivity at 300 K ( κ ph,300K ) by up to 17%. The reduction of κ ph,300K is attributed to a decrease in the specific heat and phonon relaxation time through heavy element substitution. We found that increasing the Pt content reduced the specific heat at high temperatures, which may be caused by the locked state of phasons. The observed glass-like low values of κ ph,300K (0.9-1.1 W m -1 K - 1 at 300 K) for Al 63 Cu 25- x (Au,Pt) x Fe 12 are close to the lower limit calculated using the Cahill model.

Trichoderma reesei has 11 putative β-glucosidases in its genome, playing key parts in the induction and production of cellulase. Nevertheless, the reason why the T. reesei genome encodes so many β-glucosidases and the distinct role each β-glucosidase plays in cellulase production remain unknown. In the present study, the cellular function and distribution of 10 known β-glucosidases (CEL3B, CEL3E, CEL3F, CEL3H, CEL3J, CEL1A, CEL3C, CEL1B, CEL3G, and CEL3D) were explored in T. reesei , leaving out BGL1 (CEL3A), which has been well investigated. We found that the overexpression of cel3b or cel3g significantly enhanced extracellular β-glucosidase production, whereas the overexpression of cel1b severely inhibited cellulase production by cellulose, resulting in nearly no growth of T. reesei Four types of cellular distribution patterns were observed for β-glucosidases in T. reesei : (i) CEL3B, CEL3E, CEL3F, and CEL3G forming clearly separated protein secretion vesicles in the cytoplasm; (ii) CEL3H and CEL3J diffusing the whole endomembrane as well as the cell membrane with protein aggregation, like a reticular network; (iii) CEL1A and CEL3D in vacuoles; (iv) and CEL3C in the nucleus. β-glucosidases CEL1A, CEL3B, CEL3E, CEL3F, CEL3G, CEL3H, and CEL3J were identified as extracellular, CEL3C and CEL3D as intracellular, and CEL1B as unknown. The extracellular β-glucosidases CEL3B, CEL3E, CEL3F, CEL3H, and CEL3G were secreted through a tip-directed conventional secretion pathway, and CEL1A, via a vacuole-mediated pathway that was achieved without any signal peptide, while CEL3J was secreted via an unconventional protein pathway bypassing the endoplasmic reticulum (ER) and Golgi. IMPORTANCE Although β-glucosidases play an important role in fungal cellulase induction and production, our current understanding does not provide a global perspective on β-glucosidase function. This work comprehensively studies all the β-glucosidases regarding their effect on cellulase production and their cellular distribution and secretion. Overexpression of cel3b or cel3g significantly enhanced β-glucosidase production, whereas overexpression of cel1b severely inhibited cellulase production on cellulose. In addition, overexpression of cel3b , cel3e , cel3f , cel3h , cel3j , cel3c , or cel3g delayed endoglucanase (EG) production. We first identified four cellular distribution patterns of β-glucosidases in Trichoderma reesei Specially, CEL3C was located in the nucleus. CEL3J was secreted through the nonclassical protein secretion pathway bypassing endoplasmic reticulum (ER) and Golgi. CEL1A was secreted via a vacuole-mediated conventional secretion route without a signal peptide. These findings advance our understanding of β-glucosidase properties and secretory pathways in filamentous fungi, holding key clues for future study., (Copyright © 2021 Pang et al.)

Background: Knowledge with respect to regulatory systems for cellulase production is prerequisite for exploitation of such regulatory networks to increase cellulase production, improve fermentation efficiency and reduce the relevant production cost. The target of rapamycin (TOR) signaling pathway is considered as a central signaling hub coordinating eukaryotic cell growth and metabolism with environmental inputs. However, how and to what extent the TOR signaling pathway and rapamycin are involved in cellulase production remain elusive., Result: At the early fermentation stage, high-dose rapamycin (100 μM) caused a temporary inhibition effect on cellulase production, cell growth and sporulation of Trichoderma reesei RUT-C30 independently of the carbon sources, and specifically caused a tentative morphology defect in RUT-C30 grown on cellulose. On the contrary, the lipid content of T. reesei RUT-C30 was not affected by rapamycin. Accordingly, the transcriptional levels of genes involved in the cellulase production were downregulated notably with the addition of rapamycin. Although the mRNA levels of the putative rapamycin receptor trFKBP12 was upregulated significantly by rapamycin, gene trTOR (the downstream effector of the rapamycin-FKBP12 complex) and genes associated with the TOR signaling pathways were not changed markedly. With the deletion of gene trFKBP12, there is no impact of rapamycin on cellulase production, indicating that trFKBP12 mediates the observed temporary inhibition effect of rapamycin., Conclusion: Our study shows for the first time that only high-concentration rapamycin induced a transient impact on T. reesei RUT-C30 at its early cultivation stage, demonstrating T. reesei RUT-C30 is highly resistant to rapamycin, probably due to that trTOR and its related signaling pathways were not that sensitive to rapamycin. This temporary influence of rapamycin was facilitated by gene trFKBP12. These findings add to our knowledge on the roles of rapamycin and the TOR signaling pathways play in T. reesei.

A series of Au x Si 4 - cluster anions ( x = 1-4) were generated most abundantly by laser ablation of a Au 4 Si alloy target. Photoelectron spectroscopy and density functional theory (DFT) calculation of Au x Si 4 - ( x = 1-4) revealed that Au 3 Si 4 - can be viewed as an electronically closed superatom and is composed of a Si 4 unit whose three adjacent edges of a single facet are bridged by three Au atoms. Such phase-segregated structure is facilitated by aurophilic interaction between the three Au atoms and results in a large permanent dipole moment (4.43 D). DFT calculations on an electronically equivalent superatom Au 4 Si 4 predicted a new structure in which the uncoordinated Si atom of Au 3 Si 4 - is bonded by Au + . This Au 4 Si 4 is much more stable than a cubic structure previously reported and has a large HOMO-LUMO gap (1.68 eV) and a small permanent dipole moment (0.41 D).

The magnetic susceptibility of the 1/1 approximants to icosahedral quasicrystals in a series of Cd 85- x Mg x Tb 15 ( x = 5, 10, 15, 20) alloys was investigated in detail. The occurrence of antiferromagnetic (AFM) to spin-glass (SG)-like transition was noticed by increasing Mg. Transmission electron microscopy analysis evidenced a correlation between the magnetic transition and suppression of the monoclinic superlattice ordering with respect to the orientation of the Cd 4 tetrahedron at T > 100 K. The possible origins of this phenomenon were discussed in detail. The occurrence of the AFM to SG-like magnetic transition is associated with the combination of chemical disorder due to a randomized substitution of Cd with Mg and the orientational disorder of the Cd 4 tetrahedra., (© 2020 IOP Publishing Ltd.)

A systematic investigation has been performed to elucidate effects of rare earth type and structural complexity on magnetic properties of icosahedral quasicrystal (iQC) and their cubic approximants (APs) in the ternary Cd-Mg-RE (RE = Gd, Tb, Dy, Ho, Er, and Tm) systems. At low temperatures, iQCs and 2/1 APs exhibit spin-glass-like freezing for RE = Gd, Tb, Dy, and Ho, while for Er and Tm they do not show freezing behavior down to the base temperature ∼2 K. The 1/1 APs exhibit either spin-glass-like freezing or antiferromagnetic (AFM) ordering depending on their constituent Mg content. The T f values show increasing trend from iQC to 2/1 and 1/1 APs. In contrast, the absolute values of Weiss temperature for iQCs are larger than those in 2/1 and 1/1 APs, indicating that the total AFM interactions between the neighboring spins are larger in aperiodic, rather than periodic systems. Competing spin interactions originating from the long-range Ruderman-Kittel-Kasuya-Yoshida mechanism along with chemical disorder of Cd/Mg ions presumably account for the observed spin-glass-like behavior in Cd-Mg-RE iQCs and APs., (© 2020 IOP Publishing Ltd.)

Coral polyps are basic clonal biological units of reef corals. However, in vitro experimental model for long-term physiological and ecological studies has not been well developed due to the difficulty of effectively acquiring and culturing single polyps. This study developed an experimental platform based on microfluidics for culturing single coral polyps and tracing its growth state over time in the long run. The corresponding computational modeling was conducted to predict the metabolic processes under the static and dynamic conditions by coupling the mass transfer and reaction with Navier-Stokes equations. Design and fabrication of the microfluidic chip was the key to provide a constant laminar flow environment that enabled the controlled high oxygen and bicarbonate transfer for the cultivation of the single coral polyps. The single coral polyps were induced to bail out of the coral reef upon the chemical stress and cultured for more than fifteen days in the microfluidic chip. It was found that the single coral polyps in the microfluidic chip can maintain their normal metabolic process over the cultivation period, suggesting that our microfluidic platform can serve as a suitable tool to study the coral polyps by providing a controllable and suitable biological microenvironment.

Background: Lincomycin, produced by Streptomyces lincolnensis, is a lincosamide antibiotic and widely used for the treatment of the infective diseases caused by Gram-positive bacteria. The mechanisms of lincomycin biosynthesis have been deeply explored in recent years. However, the regulatory effects of LmbU that is a transcriptional regulator in lincomycin biosynthetic (lmb) gene cluster have not been fully addressed., Results: LmbU was used to search for homologous LmbU (LmbU-like) proteins in the genomes of actinobacteria, and the results showed that LmbU-like proteins are highly distributed regulators in the biosynthetic gene clusters (BGCs) of secondary metabolites or/and out of the BGCs in actinomycetes. The overexpression, inactivation and complementation of the lmbU gene indicated that LmbU positively controls lincomycin biosynthesis in S. lincolnensis. Comparative transcriptomic analysis further revealed that LmbU activates the 28 lmb genes at whole lmb cluster manner. Furthermore, LmbU represses the transcription of the non-lmb gene hpdA in the biosynthesis of L-tyrosine, the precursor of lincomycin. LmbU up-regulates nineteen non-lmb genes, which would be involved in multi-drug flux to self-resistance, nitrate and sugar transmembrane transport and utilization, and redox metabolisms., Conclusions: LmbU is a significant pleiotropic transcriptional regulator in lincomycin biosynthesis by entirely activating the lmb cluster and regulating the non-lmb genes in Streptomyces lincolnensis. Our results first revealed the pleiotropic regulatory function of LmbU, and shed new light on the transcriptional effects of LmbU-like family proteins on antibiotic biosynthesis in actinomycetes.

Background: The ureolytic bacterium Sporosarcina pasteurii is well-known for its capability of microbially induced calcite precipitation (MICP), representing a great potential in constructional engineering and material applications. However, the molecular mechanism for its biomineralization remains unresolved, as few studies were carried out., Results: The addition of urea into the culture medium provided an alkaline environment that is suitable for S. pasteurii. As compared to S. pasteurii cultivated without urea, S. pasteurii grown with urea showed faster growth and urease production, better shape, more negative surface charge and higher biomineralization ability. To survive the unfavorable growth environment due to the absence of urea, S. pasteurii up-regulated the expression of genes involved in urease production, ATPase synthesis and flagella, possibly occupying resources that can be deployed for MICP. As compared to non-mineralizing bacteria, S. pasteurii exhibited more negative cell surface charge for binding calcium ions and more robust cell structure as nucleation sites. During MICP process, the genes for ATPase synthesis in S. pasteurii was up-regulated while genes for urease production were unchanged. Interestingly, genes involved in flagella were down-regulated during MICP, which might lead to poor mobility of S. pasteurii. Meanwhile, genes in fatty acid degradation pathway were inhibited to maintain the intact cell structure found in calcite precipitation. Both weak mobility and intact cell structure are advantageous for S. pasteurii to serve as nucleation sites during MICP., Conclusions: Four factors are demonstrated to benefit the super performance of S. pasteurii in MICP. First, the good correlation of biomass growth and urease production of S. pasteurii provides sufficient biomass and urease simultaneously for improved biomineralization. Second, the highly negative cell surface charge of S. pasteurii is good for binding calcium ions. Third, the robust cell structure and fourth, the weak mobility, are key for S. pasteurii to be nucleation sites during MICP.

Intermetallic compounds have attracted research attention in catalysis because of their unique catalytic properties. Recently, a group of intermetallic compounds, referred to as Heusler alloys (X 2 YZ), has been investigated as new catalysts. In this study, catalytic properties of 14 Heusler alloys with X = Fe, Co, Ni, or Cu; Y = Ti, Mn, or Fe; Z = Al, Si, Ga, Ge, or Sn for the steam reforming of methanol were examined. Co 2 TiAl and Ni 2 TiAl alloys exhibited relatively high H 2 production rates because of the formation of fine particles via the selective oxidation of Ti. X 2 MnZ alloys exhibited high CO 2 selectivity because of a water-gas shift reaction catalyzed by using MnO that was formed during the reaction. Crystal phases, surface microstructures, and surface compositions of most alloys were changed because of the reaction, and the formation of fine particles possibly assisted in the observation of catalytic activity. Heusler alloys can be beneficial as catalyst precursors by the selection of appropriate elemental sets depending on target reactions., Competing Interests: The authors declare no competing financial interest., (Copyright © 2019 American Chemical Society.)

Background: Filamentous fungi secret hydrolytic enzymes like cellulase and hemicellulase outside the cells, serving as important scavengers of plant biomass in nature and workhorses in the enzyme industry. Unlike the extensive study on the mechanism of cellulase production in fungi, research on spatiotemporal distribution and secretion of cellulase in fungi is lacking, retarding the deeper understanding of the molecular mechanism behind the fungal cellulase production., Result: Recombinant Trichoderma reesei strains RBGL, RCBH, and RCMC were successfully constructed from T. reesei RUT-C30, expressing red fluorescent protein DsRed-tagged versions of β-glucosidase (BGL), cellobiohydrolase (CBH), and endoglucanase (CMC), respectively. With the assistance of these strains, we found that all three cellulase components BGL, CBH, and CMC diffused throughout the whole fungal mycelium with major accumulation at the hyphal apexes. These enzymes located in ER, Golgi, vacuoles and cell membrane/wall, but not septum, and secreted abundantly into the culture medium. Moreover, the major secretion of CBH and CMC started more early than that of BGL. Brefeldin A (BFA) completely blocked cellulase expression and secretion in T. reesei., Conclusion: Based on recombinant T. reesei RBGL, RCBH, and RCMC expressing DsRed-fused versions of BGL, CBH, and CMC, respectively, the distribution and secretion of cellulase production in T. reesei were first visualized directly in a dynamic way, preliminarily mapping the location and secretion of T. reesei cellulase and providing evidence for revealing the secretion pathways of cellulase in T. reesei . The obtained results suggest that cellulase excretion majorly occurs via the conventional ER-Golgi secretory pathway, and might be assisted through unconventional protein secretion pathways., Competing Interests: Competing interestsThe authors declare that they have no competing interests.

We report on the formation of a new icosahedral quasicrystal ( i QC) in the Au-Sn-Yb alloy system. This i QC has a primitive icosahedral lattice with a lattice constant a ico of 0.5447(7) nm and a composition that was determined to be Au 60.0 Sn 26.7 Yb 13.3 . X-ray absorption spectroscopy measurement of the near Yb L 3 edge demonstrates that the Yb valence in the i QC is an intermediate valence between divalent (4f 14 ) and trivalent (4f 13 ) at ambient pressure and was determined to be 2.18+. The results are compared to those for a corresponding 2/1 cubic approximant crystal. The formation of this new i QC is discussed in terms of the atomic size factor (δ) and the valence electron-to-atom ratio ( e / a ).

Hydrogen storage properties and reactivity for hydrogenation of acetylene in a series of CeNi 5- x Ga x ( x = 0, 0.5, 0.75, 1, 1.25, 1.5) alloys and Mg 2 Ni were determined and compared. The structure of CeNi 5 (CaCu 5 type) was maintained up to CeNi 3.5 Ga 1.5 when Ni was replaced by Ga. The replacement facilitated hydrogenation absorption by creating larger interstitial spaces through expansion of the lattice, allowing CeNi 4.25 Ga 0.75 to absorb the greatest proportion of hydrogen atoms among the alloys under the same conditions. The results showed that the absorbed hydrogen in CeNi 3.75 Ga 1.25 improved reactivity. In contrast, Mg 2 Ni formed a hydride upon hydrogenation of acetylene and thus possessed much lower activity. The difference of the activity of absorbed hydrogen between CeNi 5- x Ga x and Mg 2 Ni was confirmed from transient response tests under reaction gases alternately containing He and H 2 .

The Yb-Ga-Au 1/1 quasicrystal approximant (AP) composition ranges from Yb 14.0 Ga 20.6 Au 65.4 to Yb 14.8 Ga 46.3 Au 38.9 , and single crystals of the 1/1 AP having the composition Yb 13.8 Ga 26.1 Au 60.1 were obtained by the self-flux technique. X-ray structural analysis demonstrated that the atomic structure [space group Im3; a = 14.6889(9) Å] can be described by the body-centered packing of Tsai-type rhombic triacontahedron (RTH) clusters. The positional disorder in these clusters, interpreted as the average of an orientationally disordered tetrahedron and triangle, results in positional disorder in the outer shells. The elemental distributions and positions of mixtures of Au and Ga atoms in the RTH clusters correspond to those in the isostructural Yb 15 Al 36 Au 49 1/1 AP.

CRISPR utilizing Cas9 from Streptococcus pyogenes (SpCas9) and CRISPR interference (CRISPRi) employing catalytically inactive SpCas9 (SpdCas9) have gained popularity for Escherichia coli engineering. To integrate the SpdCas9-based CRISPRi module using CRISPR while avoiding mutual interference between SpCas9/SpdCas9 and their cognate single-guide RNA (sgRNA), this study aimed at exploring an alternative Cas nuclease orthogonal to SpCas9. We compared several Cas9 variants from different microorganisms such as Staphylococcus aureus (SaCas9) and Streptococcus thermophilius CRISPR1 (St1Cas9) as well as Cas12a derived from Francisella novicida (FnCas12a). At the commonly used E. coli model genes  LacZ, we found that SaCas9 and St1Cas9 induced DNA cleavage more effectively than FnCas12a. Both St1Cas9 and SaCas9 were orthogonal to SpCas9 and the induced DNA cleavage promoted the integration of heterologous DNA of up to 10 kb, at which size St1Cas9 was superior to SaCas9 in recombination frequency/accuracy. We harnessed the St1Cas9 system to integrate SpdCas9 and sgRNA arrays for constitutive knockdown of three genes, knock-in pyc and knockout adhE, without compromising the CRISPRi knockdown efficiency. The combination of orthogonal CRISPR/CRISPRi for metabolic engineering enhanced succinate production while inhibiting byproduct formation and may pave a new avenue to E. coli engineering., (© 2019 Wiley Periodicals, Inc.)

The phase and local environment, neighbouring atoms and coordination numbers (CN), for an Al-Cu-Fe multilayer were studied during heating (to 800 °C) and cooling (to room temperature) processes using in-situ X-Ray diffraction (XRD) and in-situ X-ray absorption spectroscopy (XAS) techniques to investigate the formation of Al-Cu-Fe quasicrystals (QCs). In-situ XRD clarified the transition of the ω-Al 7 Cu 2 Fe phase to a liquid state at the high temperature which transformed into the QC phase during cooling. The in-situ XAS showed a relatively small shift in distance between Cu-Al and Fe-Al during the phase evolution from RT to 700 °C. The distance between Cu-Cu, however, showed a significant increase from ω-phase at 700 °C to the liquid state at 800 °C, and this distance was maintained after QC formation. Furthermore, the CN of Fe-Al was changed to N = 9 during cooling. Through our observations of changes in CN, atomic distances and the atomic environment, we propose the local structural ordering of the quasicrystalline phase originated from a liquid state via ω-phase. In this study, we give a clear picture of the atomic environment from the crystalline to the quasicrystalline phase during the phase transitions, which provides a better understanding of the synthesis of functional QC nanomaterials.

Heusler alloys ( X 2 YZ ) are well-established intermetallic compound materials in various fields because their function can be precisely adjusted by elemental substitution (e.g., X 2 YZ 1- x Z ' x ). Although intermetallic compound catalysts started attracting attention recently, catalysis researchers are not familiar with Heusler alloys. We report their potential as novel catalysts focusing on the selective hydrogenation of alkynes. We found that Co 2 MnGe and Co 2 FeGe alloys have great alkene selectivity. Mutual substitution of Mn and Fe (Co 2 Mn x Fe 1- x Ge) enhanced the reaction rate without changing selectivity. The substitution of Ga for Ge decreased the selectivity but increased the reaction rate monotonically with Ga composition. Elucidation of these mechanisms revealed that the fine tuning of catalytic properties is possible in Heusler alloys by separately using ligand and ensemble effects of elemental substitution.

Human has greatly disturbed nitrogen (N) cycles through the food production and consumption (FPC) activities, which caused serious environmental consequences. A few studies address the pathways of N cycles to clarify the interrelationships between human activities and the altered N cycles, especially in terms of the spatial aspects of N flows, which are also influenced by natural factors. Here, we developed an integrated model to quantify the spatiotemporal patterns of N flows within the FPC and applied it to the Chaohu Watershed from 1949 to 2012. We found that total N inputs into the FPC increased by 6-fold during 1949-2012, while N inputs from chemical fertilizer had the largest increase, with an annual growth rate of 8.6%. N generation in manure doubled, but the recycling rate decreased by 50% in the study period. The total N losses increased from 17 to 130Gg-N/yr between 1949 and 2012. Annually, ~7% of total N losses was discharged into surface water annually, which peaked at 9.5Gg-N/yr during 1998-2003, two times as that in the early 1980s when algae blooms first occurred in Chaohu lake. Spatially, around one-fifth of the catchment area was responsible for all of N losses to surface water. These intensive areas located in the eastern and southern-central regions, which were identified as the critical areas for priority N mitigation. This study provided a method to depict spatiotemporal patterns of regional N flows for identifying the critical sources and areas of N pollution for precision watershed management., (Copyright © 2017 Elsevier B.V. All rights reserved.)

The atomic structure of a 0.2 atom % Pt-doped complex metallic alloy, monoclinic Al 13 Fe 4 , was investigated using a single crystal prepared by the Czochralski method. High-angle annular dark-field scanning transmission electron microscopy showed that the Pt atoms were dispersed as single atoms and substituted at Fe sites in Al 13 Fe 4 . Single-crystal X-ray structural analysis revealed that the Pt atoms preferentially substitute at Fe(1). Unlike those that have been reported, Pt single atoms in the surface layers showed lower activity and selectivity than those of Al 2 Pt and bulk Pt for propyne hydrogenation, indicating that the active state of a given single-atom Pt site is strongly dominated by the bonding to surrounding Al atoms.

The fcc lattice of porous Cu prepared by dealloying Al 2 Cu with HCl aqueous solution exhibits a high density of twinning defects with an average domain size of about 3 nm along the ⟨111⟩ directions. The high density of twinning was verified by X-ray diffraction and qualitatively interpreted by a structural model showing the 5% probability of twinning defect formation. Most of the twinning defects disappeared after annealing at 873 K for 24 h. Twinned Cu reveals much faster oxidation rate in comparison to that without (or with much fewer) twinning defects, as shown by X-ray diffraction and hydrogen differential scanning calorimetry. Using ab initio DFT calculations, we demonstrate that twinning defects in porous Cu are able to form nucleation centers for the growth of Cu 2 O. The geometry of the V-shaped edges on the twinned {211} surfaces is favorable for formation of the basic structural elements of Cu 2 O. The fast oxidation of porous Cu prepared by dealloying can thus be explained by the fast formation of the Cu 2 O nucleation centers and their high density.

Monensin, a polyether ionophore antibiotic, is produced by Streptomyces cinnamonensis and worldwide used as a coccidiostat and growth-promoting agent in the field of animal feeding. The monensin biosynthetic gene cluster (mon) has been reported. In this study, the potential functions of three putatively pathway-specific regulators (MonH, MonRI, and MonRII) were clarified. The results from gene inactivation, complementation, and overexpression showed that MonH, MonRI, and MonRII positively regulate monensin production. Both MonH and MonRI are essential for monensin biosynthesis, while MonRII is non-essential and could be completely replaced by additional expression of monRI. Transcriptional analysis of the mon cluster by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and electrophoresis mobility shift assays (EMSAs) revealed a co-regulatory cascade process. MonH upregulates the transcription of monRII, and MonRII in turn enhances the transcription of monRI. MonRII is an autorepressor, while MonRI is an autoactivator. MonH activates the transcription of monCII-monE, and upregulates the transcription of monT that is repressed by MonRII. monAX and monD are activated by MonRI, and upregulated by MonRII. Co-regulation of those post-polyketide synthase (post-PKS) genes by MonH, MonRI, and MonRII would contribute to high production of monensin. These results shed new light on the transcriptional regulatory cascades of antibiotic biosynthesis in Streptomyces.

Nanoporous gold (NPG) prepared by de-alloying Al 2 Au exhibits correlation between the high catalytic reactivity towards CO oxidation and the density of twinning defects in the fcc lattice of NPG. It was also discovered that on the internal surface of NPG, quite common twinning defects can create close-packed rows of six-coordinated catalytically active Au atoms denoted as W-chains. In this work, using density functional theory methods, we investigate energy conditions for formation, thermal stability, and chemical reactivity of these active sites. The possibility of dioxygen chemisorption on various surface sites is studied in detail. A contribution from the dispersion interactions is also considered. The calculated surface density of the active six-coordinated atoms in NPG comparable with that of supported gold nanoparticle catalysts, exothermic chemisorption of dioxygen, and the energy profiles of reaction pathways for CO oxidation indicate that the six-coordinated sites created by twinning can significantly contribute to the catalytic activity of NPG.

In this study, we investigated the catalytic properties of various Heusler alloys for the hydrogenation of propyne and the oxidation of carbon monoxide. For propyne hydrogenation, Co 2 FeGe alloy showed a higher activity than that of elemental Co, where neither Fe nor Ge showed any activity. This clearly indicates an alloying effect. For the oxidation of carbon monoxide, although most alloys showed a significant change in catalytic activity during measurement due to an irreversible oxidation of the alloy, Co 2 TiSn alloy showed a very small change. The results indicate that the catalytic activity and stability of a Heusler alloy can be tuned by employing an appropriate set of elements., Competing Interests: The authors declare no competing financial interest.

To accelerate healing of severe hemorrhagic wounds, a novel highly absorbent hemostatic dressing composed of a Tencel ® /absorbent-cotton/polylactic acid nonwoven base and chitosan/nanosilver antibacterial agent was fabricated by using a nonwoven processing technique and a freeze-drying technique. This study is the first to investigate the wicking and water-absorbing properties of a nonwoven base by measuring the vertical wicking height and water absorption ratio. Moreover, blood agglutination and hemostatic second tests were conducted to evaluate the hemostatic performance of the resultant wound dressing. The blending ratio of fibers, areal weight, punching density, and fiber orientation, all significantly influenced the vertical moisture wicking property. However, only the first two parameters markedly affected the water absorption ratio. After the nonwoven base absorbed blood, scanning electron microscope (SEM) observation showed that erythrocytes were trapped between the fibrin/clot network and nonwoven fibers when coagulation pathways were activated. Prothrombin time (PT) and activated partial thromboplastin time (APTT) blood agglutination of the resultant dressing decreased to 14.34 and 50.94 s, respectively. In the femoral artery of the rate bleeding model, hemostatic time was saved by 87.2% compared with that of cotton cloth. Therefore, the resultant antibacterial wound dressing demonstrated greater water and blood absorption, as well as hemostatic performance, than the commercially available cotton cloth, especially for healing severe hemorrhagic wounds.

We describe a new mechanism for creation of catalytically active sites in porous gold. Samples of porous gold prepared by de-alloying Al2Au exhibit a clear correlation between the catalytic reactivity towards CO oxidation and structural defects in the fcc lattice of Au. We have found that on the stepped {211} surfaces quite common twin boundary defects in the bulk structure of porous gold can form long close-packed rows of atoms with the coordination number CN = 6. DFT calculations confirm that on these low-coordinated Au sites dioxygen chemisorbs and CO oxidation can proceed via the Langmuir-Hinshelwood mechanism with the activation energy of 37 kJ/mol or via the CO-OO intermediate with the energy barrier of 19 kJ/mol. The existence of the twins in porous gold is stabilized by the surface energy.

Chemical modifications of DNA comprise epigenetic mechanisms that contribute to the maintenance of cellular activities and memory. Although the function of 5-methylcytosine (5-mC) has been extensively studied, little is known about the function(s) of relatively rarer and underappreciated cytosine modifications including 5-hydroxymethylcytosine (5-hmC). The discovery that ten-eleven translocation (Tet) proteins mediate conversion of 5-mC to 5-hmC, and other oxidation derivatives, sparked renewed interest to understand the biological role of 5-hmC. Studies examining total 5-hmC levels revealed the highly dynamic yet tissue-specific nature of this modification, implicating a role in epigenetic regulation and development. Intriguingly, 5-hmC levels are highest during early development and in the brain where abnormal patterns of 5-hmC have been observed in disease conditions. Thus, 5-hmC adds to the growing list of epigenetic modifications with potential utility in clinical applications and warrants further investigation. This review discusses the emerging functional roles of 5-hmC in normal and disease states, focusing primarily on insights provided by recent studies exploring the genome-wide distribution of this modification in mammals.

Three different porous Au catalysts that exhibit high catalytic activity for CO oxidation were prepared by the leaching of Al from an intermetallic compound, Al2Au, with 10 wt. %-NaOH, HNO3, or HCl aqueous solutions. The catalysts were investigated using Brunauer-Emmett-Teller measurements, synchrotron X-ray powder diffraction, hard X-ray photoelectron spectroscopy, field emission scanning electron microscopy, and transmission electron microscopy (TEM). Broad diffraction peaks generated during the leaching process correlated with high activity for all the porous Au catalysts. CO oxidation catalyzed by porous Au leached with NaOH and HNO3 is considered to be dominated by different mechanisms at low (< 320 K) and high (> 370 K) temperatures. Activity in the low-temperature region is mainly attributed to the perimeter interface between residual Al species (AlOx) and porous Au, whereas activity in the high-temperature region results from a high density of lattice defects such as twins and dislocations, which were evident from diffraction peak broadening and were observed with high-resolution TEM in the porous Au leached with NaOH. It is proposed that atoms located at lattice defects on the surfaces of porous Au are the active sites for catalytic reactions.

Aims: To improve lincomycin A production and decrease the content of byproduct lincomycin B in an industrial lincomycin-producing strain., Methods and Results: The in silico analysis indicated that LmbW could be involved in propylproline biosynthesis of lincomyin A. In this study, we constructed an lmbW deletion mutant and found that the mutant lost the ability to produce lincomycin A, but increased the accumulation of lincomycin B. The loss of lincomycin A production can be restored by complementing the mutant with the expression of lmbW gene. When lmbW and metK (encoding S-adenosylmethionine synthetase) was co-overexpressed, lincomycin A titre was 1744·6 mg l(-1) , a 35·83% improvement over the original strain. Meanwhile, the content of lincomycin B was reduced to 4·41%, a remarkable decrease of 34·76%, compared to that of the original strain., Conclusions: lmbW encodes a C-methyltransferase involved in the biosynthesis of lincomycin A but not lincomycin B. Co-overexpression of lmbW and metK improved lincomycin A production and decreased the content of lincomycin B., Significance and Impact of the Study: The engineered Streptomyces lincolnensis strain shows promising application in the fermentation production of lincomycin A, which may help cut production costs and simplify downstream separation processes., (© 2015 The Society for Applied Microbiology.)