Search

Human hyaluronidase 1 (HYAL1) and PH20 play vital roles in degrading hyaluronic acids through the substrate-assisted double displacement mechanism. While HYAL1, a lysosomal enzyme, functions optimally under acidic conditions, PH20, a sperm surface hyaluronidase, displays a broader pH range, from acidic to neutral. Our objective was to extend HYAL1's pH range towards neutral pH by introducing repulsive charge-charge interactions involving the catalytic Glu131, increasing its pKa as the proton donor. Substituting individual acidic residues in the β3-loop (S77D), β3'-β3″ hairpin (T86D and P87E), and at Ala132 (A132D and A132E) enabled HYAL1 to demonstrate enzyme activity at pH 7, with the mutants S77D, P87E, and A132E showing the highest activity in the substrate gel assay. However, double and triple substitutions, including S77D/T86D/A132E as found in the PH20 configuration, did not result in enhanced activity compared to single substitutions. Conversely, PH20 mutants with non-acidic substitutions, such as D94S in the β3-loop and D103T in the β3'-β3″ hairpin, significantly reduced activity within the pH range of 4 to 7. However, the PH20 mutant E149A, reciprocally substituted compared to A132E in HYAL1, exhibited activity similar to PH20 wild-type (WT) at pH 7. In a turbidimetric assay, HYAL1 mutants with single acidic substitutions exhibited activity similar to that of PH20 WT at pH 7. These results suggest that substituting acidic residues near Glu131 results in HYAL1 activity at neutral pH through electrostatic repulsion. This study highlights the significance of charge-charge interactions in both HYAL1 and PH20 in regulating the pH-dependent activity of hyaluronidases.

AbstractWe introduce dGSLM, the first “textless” model able to generate audio samples of naturalistic spoken dialogues. It uses recent work on unsupervised spoken unit discovery coupled with a dual-tower transformer architecture with cross-attention trained on 2000 hours of two-channel raw conversational audio (Fisher dataset) without any text or labels. We show that our model is able to generate speech, laughter, and other paralinguistic signals in the two channels simultaneously and reproduces more naturalistic and fluid turn taking compared to a text-based cascaded model.1,2

In this study, ZnO/g-C3N4 nanocomposites were prepared via a physical mixing-calcination process for improved degradation of dyes under visible light irradiation. The BET surface area, pore volume, crystal size, and pHpzc of the ZnO/g-C3N4 composite were 3.9 m2/g, 0.034 cm3/g, 18.1 nm, and 7.7, respectively. Although the morphology of the ZnO/g-C3N4 composite was very different from that of pure g-C3N4, their average pore sizes were similar. The Eg of the ZnO/g-C3N4 composite (3.195 eV) was slightly lower than that of ZnO (3.195) but much higher than that of g-C3N4 (2.875). The interface interaction of ZnO and g-C3N4, which was revealed by oscillations of Zn-C, benefited the transport of photoinduced charge carriers and reduced the recombination of electron-hole. As the result, the ZnO/g-C3N4 composite had higher photocatalytic activity than ZnO and g-C3N4. Its degradation efficiency (DE) value for methylene blue (MB) in 90 min and rate constant were 93.2 % and 0.025 min‑1, respectively. In addition, the effects of ZnO/urea molar ratio, catalyst dosage, solution pH, and concentration of dye on photocatalytic degradation of MB were completely investigated. The photocatalytic performance of the ZnO/g-C3N4 composite was evaluated by the degradation of other persistent organic compounds, also compared to other catalysts in the literatures. Copyright © 2022 by Authors, Published by BCREC Group. This is an open access article under the CC BY-SA License (https://creativecommons.org/licenses/by-sa/4.0).

The Spitzenkörper (SPK) is a polarized accumulation of proteins and secretory vesicles associated with tip growth of fungal hyphae. Here, Zheng et al. study SPK assembly and dynamics, identify SPK protein scaffolds and associated proteins, and reveal similarities with other scaffolds from metazoans.

The fungal Ascomycota provide a model phylum to investigate the evolution of complex multicellularity. Here, the authors combine genome sequencing with comparative and functional genomics to identify diverse endomembrane related machineries associated with the gain and loss of fungal complexity.

Horizontal gene transfer (HGT) can promote evolutionary adaptation by transforming a species' relationship to the environment. In most well-understood cases of HGT, acquired and donor functions appear to remain closely related. Thus, the degree to which HGT can lead to evolutionary novelties remains unclear. Mucorales fungi sense gravity through the sedimentation of vacuolar protein crystals. Here, we identify the octahedral crystal matrix protein (OCTIN). Phylogenetic analysis strongly supports acquisition of octin by HGT from bacteria. A bacterial OCTIN forms high-order periplasmic oligomers, and inter-molecular disulphide bonds are formed by both fungal and bacterial OCTINs, suggesting that they share elements of a conserved assembly mechanism. However, estimated sedimentation velocities preclude a gravity-sensing function for the bacterial structures. Together, our data suggest that HGT from bacteria into the Mucorales allowed a dramatic increase in assembly scale and emergence of the gravity-sensing function. We conclude that HGT can lead to evolutionary novelties that emerge depending on the physiological and cellular context of protein assembly.

Restructuring in Vietnam has become a critical necessity to ensure efficiency and stability, especially in the context of long-lasting recession in the economy as a whole. This paper reviews Vietnam’s economic growth model to point out the demand for restructuring the key blocks of the economy. The paper also discusses the road map for Vietnam’s restructuring process and synthesizes the initial achievements of the process so far.

Nowadays, the implementation and application of the BIM process in construction project management is a pressing need. This aligns with the global development trends in the construction sector and project information management in general. Numerous scholars and companies are actively engaged in learning, understanding, and investigating various aspects of BIM to stay up-to-date and meet the inevitable developmental requirements. This study focuses on the role and application of BIM, intending to identify limitations that hinder its fulfillment of expectations in project schedule management. In addition, it explores studies that show how other countries have effectively employed BIM in project management and progress tracking throughout the project lifecycle. The study aims to address three main objectives: (a) comprehensively examine and provide evidence related to the concept of BIM in project schedule management, (b) present the benefits of applying BIM in comparison to traditional methods in project management and operation, and (c) identify limitations stemming from various factors that may pose challenges in the application of BIM in project schedule management. [ABSTRACT FROM AUTHOR]

Waste frying oil (WFO) can take a heavy toll on people' health and the environment; however, they can still be useful in reality if they are appropriately collected and recycled. Specifically, we can take their advantages to create a nutritious carbon source for bacteria growth. In this research, due to a strong desire to enhance the value of such products, Pseudomonas putida KT2440 is employed to produce polyhydroxyalkanoates (PHA) which is a group of natural biodegradable polyesters synthesized by microorganisms. Two deficient glucose assimilation chassis strains, which are the glk and gcd created through knock-out genes encoding glucose dehydrogenase (gcd) and glucoskinase (glk) respectively, are compared with P.putida KT2440 WildType (WT) for optimization of PHA production. Batch cultivation coupled with an approach of standard submerged fermentation is used in order to monitor the biosynthesis of PHA and the growth profile of producer strains. The results indicate a significant improvement of PHA yields when using WFO at a concentration of 20 g/L and applying gene knock-outs, especiallyglk. Although there are still drawbacks, our results basically proclaim that the amount of gained PHA by using WFO is higher than glycerol which is the material of choice for PHA production. Additionally, the successful incorporation of PHA with Tylosin, an antibiotic belonging to the macrolide group, could be the fresh direction in fights against antibiotics resistance. To sum up, P. putida can convert greasy wastes into biopolymers boasting wide applications in diverse fields, which paves the way to develop the straightforward procedure to recycle and value this waste on a real life. [ABSTRACT FROM AUTHOR]

Hydrophobic interactions and hydrogen bonds are 2 types of noncovalent interactions that play distinct roles in the folding and structural stability of proteins. However, the specific roles of these interactions in hydrophobic or hydrophilic environments in α/β-hydrolases are not fully understood. A hyperthermophilic esterase EstE1 in a dimer maintains the C-terminal β8-α9 strand-helix via hydrophobic interactions (Phe276 and Leu299), constituting a closed dimer interface. Moreover, a mesophilic esterase rPPE in a monomer maintains the same strand-helix via a hydrogen bond (Tyr281 and Gln306). Unpaired polar residues (F276Y in EstE1 and Y281A/F and Q306A in rPPE) or reduced hydrophobic interactions (F276A/L299A in EstE1) between the β8-α9 strand-helix decrease thermal stability. EstE1 (F276Y/L299Q) and rPPE WT, both with the β8-α9 hydrogen bond, showed the same thermal stability as EstE1 WT and rPPE (Y281F/Q306L), which possess hydrophobic interactions instead. However, EstE1 (F276Y/L299Q) and rPPE WT exhibited higher enzymatic activity than EstE1 WT and rPPE (Y281F/Q306L), respectively. This suggests that α/β-hydrolases favor the β8-α9 hydrogen bond for catalytic activity in monomers or oligomers. Overall, these findings demonstrate how α/β-hydrolases modulate hydrophobic interactions and hydrogen bonds to adapt to different environments. Both types of interactions contribute equally to thermal stability, but the hydrogen bond is preferred for catalytic activity. IMPORTANCE Esterases hydrolyze short to medium-chain monoesters and contain a catalytic His on a loop between the C-terminal β8-strand and α9-helix. This study explores how hyperthermophilic esterase EstE1 and mesophilic esterase rPPE adapt to different temperatures by utilizing the β8-α9 hydrogen bonds or hydrophobic interactions differently. EstE1 forms a hydrophobic dimer interface, while rPPE forms a monomer stabilized by a hydrogen bond. The study demonstrates that these enzymes stabilize β8-α9 strand-helix differently but achieve similar thermal stability. While the β8-α9 hydrogen bond or hydrophobic interactions contribute equally to thermal stability, the hydrogen bond provides higher activity due to increased catalytic His loop flexibility in both EstE1 and rPPE. These findings reveal how enzymes adapt to extreme environments while maintaining their functions and have implications for engineering enzymes with desired activities and stabilities. [ABSTRACT FROM AUTHOR]

In this study, ZnO/g-C3N4 nanocomposites were prepared via a physical mixing-calcination process for improved degradation of dyes under visible light irradiation. The BET surface area, pore volume, crystal size, and pHpzc of the ZnO/g-C3N4 composite were 3.9 m2/g, 0.034 cm3/g, 18.1 nm, and 7.7, respectively. Although the morphology of the ZnO/g-C3N4 composite was very different from that of pure g-C3N4, their average pore sizes were similar. The Eg of the ZnO/g-C3N4 composite (3.195 eV) was slightly lower than that of ZnO (3.195) but much higher than that of g-C3N4 (2.875). The interface interaction of ZnO and g-C3N4, which was revealed by oscillations of Zn-C, benefited the transport of photoinduced charge carriers and reduced the recombination of electron-hole. As the result, the ZnO/g-C3N4 composite had higher photocatalytic activity than ZnO and g-C3N4. Its degradation efficiency (DE) value for methylene blue (MB) in 90 min and rate constant were 93.2 % and 0.025 min-1, respectively. In addition, the effects of ZnO/urea molar ratio, catalyst dosage, solution pH, and concentration of dye on photocatalytic degradation of MB were completely investigated. The photocatalytic performance of the ZnO/g-C3N4 composite was evaluated by the degradation of other persistent organic compounds, also compared to other catalysts in the literatures. [ABSTRACT FROM AUTHOR]

This study evaluates the technical feasibility of using microwave radiation for the rapid treatment of human feces. Human feces of 1000 g were radiated with a commercially available household microwave oven (with rotation) at different exposure time lengths (30, 50, 60, 70, and 75 mins) and powers (600, 800, and 1000 W). Volume reduction over 90% occurred after 1000 W microwave radiation for 75 mins. Pathogen eradiation performances of six log units or more at a high range of microwave powers were achieved. Treatments with the same energy input of 1000 Wh, but at lower powers with prolonged exposure times, significantly enhanced moisture removal and volume reduction. Microwave radiation caused carbonization and resulted in a more stable end product. The energy content of the samples after microwave treatment at 1000 W and 75 mins is 3517 ± 8.85 calories/g of dried sample, and the product can also be used as compost. [ABSTRACT FROM AUTHOR]

Study Objective: To demonstrate the feasibility of robotic single-site resection of advanced endometriosis using new technology.Design: We show a video that demonstrates our technique for accomplishing single-site laparoscopic resection of advanced endometriosis. The video is a step-by-step explanation of robotic single-site resection of endometriosis nodules overlying the ureter and rectum.Background: Laparoscopic surgery has been shown to effectively improve pain and fertility in women with endometriosis [1]. Compared with traditional multiport laparoscopy, single-incision laparoscopy is associated with similar incidence rates of blood loss, conversion to open laparotomy, and wound complications, but it has superior cosmetic outcomes and high patient satisfaction [2-5]. Furthermore, robotic single-incision laparoscopy combined with robotic Firefly technology potentially increases the removal of invisible endometriosis. Without complete resection of endometriosis, patients are less likely to achieve full pain relief postoperatively.Setting: University hospital.Patient: A 36 year old G1P1 female was referred for chronic pelvic pain. She described her pain as hip pain, pain with walking, dyspareunia, dyschezia and right anterior abdominal wall pain.Intervention: To improve detection of endometriosis, we injected the patient with indocyanine green (ICG), a fluorescent dye with widespread medical applications in identifying increased vascularity of tissues. We then visualized the tissues with robotic Firefly technology, a fluorescence-detection tool built into the da Vinci SI Surgical Systems (Intuitive Surgical, Inc, Sunnyvale, CA).Main Results: Because endometriosis lesions are associated with a high degree of neovascularization, the ICG turned the endometriosis tissues dark green, thereby enabling us to detect endometriosis that would not have been seen as readily with conventional single-site laparoscopy. This video demonstrates our technique for successfully accomplishing a single-site laparoscopic resection of advanced endometriosis, including ureterolysis, adhesiolysis, peritoneal stripping, and a rectal nodule excision.Conclusions: We found that Firefly technology and ICG facilitated identification of endometriosis in single-site robotic surgery. We were able to successfully perform single-site laparoscopic resection of advanced endometriosis nodules overlying the ureter and rectum with complete resolution of pelvic pain symptoms and excellent cosmetic results. [ABSTRACT FROM AUTHOR]

AbstractWe introduce Generative Spoken Language Modeling, the task of learning the acoustic and linguistic characteristics of a language from raw audio (no text, no labels), and a set of metrics to automatically evaluate the learned representations at acoustic and linguistic levels for both encoding and generation. We set up baseline systems consisting of a discrete speech encoder (returning pseudo-text units), a generative language model (trained on pseudo- text), and a speech decoder (generating a waveform from pseudo-text) all trained without supervision and validate the proposed metrics with human evaluation. Across 3 speech encoders (CPC, wav2vec 2.0, HuBERT), we find that the number of discrete units (50, 100, or 200) matters in a task-dependent and encoder- dependent way, and that some combinations approach text-based systems.1

The mining of frequent weighted patterns (FWPs) that considers the different semantic significance (weight) of items is more suitable for practice than the mining of frequent patterns. Therefore, it plays a vital role in real-world scenarios. However, there exist several limitations when applying methods for mining FWPs designed for static data on growth datasets, especially data streams. Hence, this study proposes an algorithm for mining FWPs over data streams. First, we introduce the concept of mining FWPs over data streams via a sliding window model. Then, we introduce a modification of the weighted node tree (WN-tree) named SWN-tree that has the ability to maintain the information over data streams. Next, this study develops a method for mining FWPs over data streams employing a sliding window model based on SWN-tree. This method is called FWPODS (Frequent Weighted Patterns Over Data Stream) algorithm. Finally, we conduct empirical experiments to compare the performances of our approach and the state-of-the-art algorithm (NFWI) for mining FWPs over data streams. The results of experiment indicate that our approach outperforms the NFWI algorithm when running in batch mode in a sliding window.