As a virulence factor, HupB plays important roles in the survival of MTB after infection and modulates the host immune response. In the current study, we aim to explore a new cellular immunological ...detection method for tuberculosis infection detection based on HupB protein.
HupB was used to stimulate PBMCs extracted from pulmonary tuberculosis (PTB) patients, and secreted cytokines was examined. Then, we constructed a single center and a multi-center clinical trials to collect PBMCs from PTB patients, nPTB patients, or healthy volunteers to verify our findings.
Cytokine's screening illustrated that IL-6 was the only cytokine released after HupB stimulation. Single-center and multi-center clinical trials showed that HupB stimulation significantly increased the level of IL-6 in the supernatant of PBMCs from PTB patients. Then we compared the specificity and sensitivity of HupB induced IL-6 release assay with ESAT-6 and CFP10 induced interferon γ release assay (IGRA), and found in smear positive PTB patients, the specificity and sensitivity of HupB induced IL-6 release assay was better than IGRA, and in smear negative PTB patients, the sensitivity was better. Combination of both assays provided an improved specificity and sensitivity for tuberculosis diagnosis.
This study explored an immunological detection method for tuberculosis infection cells based on HupB protein-induced IL-6 release test, which can be used to enhance the diagnosis diagnostic accuracy of TB.
Inspired by the catalytic mechanism of cytochrome P450 enzymes, we have evolved a fatty acid hydroxylase, P450BSβ, into a decarboxylase after four rounds of saturation mutagenesis that now ...predominantly converts fatty acids to 1-alkenes. The engineered enzyme directly utilizes inexpensive H2O2 to oxidatively decarboxylate C6–C18 saturated fatty acids as well as plant oil-derived unsaturated fatty acids, displaying high reactivity (e.g., 3,700 h–1 of turnover frequency for myristic acid). With in situ H2O2 supplied by glucose oxidase, this enzyme delivers 1-alkenes in a more productive and scalable fashion (up to 99% yield). Our directed evolution efforts present a strategy for developing new fatty acid decarboxylase for downstream biocatalytic applications.
Herein, we report an engineered enzyme that can monooxygenate unprotected tryptophan into the corresponding 3a‐hydroxyhexahydropyrrolo2,3‐bindole‐2‐carboxylic acid (HPIC) in a single, scalable step ...with excellent turnover number and diastereoselectivity. Taking advantage of directed evolution, we analyzed the stepwise oxygen‐insertion mechanism of tryptophan 2,3‐dioxygenases, and transformed tryptophan 2,3‐dioxygenase from Xanthomonas campestris into a monooxygenase for oxidative cyclization of tryptophans. It was revealed that residue F51 is vital in determining the product ratio of HPIC to N′‐formylkynurenine. Our reactions and purification procedures use no organic solvents, resulting in an eco‐friendly method to prepare HPICs for further applications.
Directed evolution: The tryptophan 2,3‐dioxygenase from Xanthomonas campestris (xcTDO) was transformed into a monooxygenase using directed evolution. The engineered xcTDO exhibits an excellent turnover number and high diastereoselectivity, and enables the scalable synthesis of 3a‐hydroxyhexahydropyrrolo2,3‐bindole‐2‐carboxylic acids (HPICs) from tryptophans in one step.
The insect order Blattodea (cockroaches and termites) has drawn substantial research attention for their dietary habits and lifestyle of living with or around humans. In the present study, we focused ...on the discovery of RNA viruses hidden in Blattodea insects using the publicly available RNA sequencing datasets. Overall, 136 distinctive RNA viruses were identified from 36 Blattodea species, of which more than 70 % were most closely related to the invertebrate-associated viral groups within Picornavirales , Sobelivirales , Bunyaviricetes , Jingchuvirales , Durnavirales , Lispiviridae , Orthomyxoviridae , Permutotetraviridae , Flaviviridae and Muvirales . Several viruses were associated with pathogens of vertebrates ( Paramyxoviridae ), plants ( Tymovirales ), protozoa ( Totiviridae ), fungi ( Narnaviridae ) and bacteria ( Norzivirales ). Collectively, 93 complete or near-complete viral genomes were retrieved from the datasets, and several viruses appeared to have remarkable temporal and spatial distributions. Interestingly, the newly identified Periplaneta americana dicistrovirus displayed a remarkable distinct bicistronic genome arrangement from the well-recognized dicistroviruses with the translocated structural and non-structural polyprotein encoding open reading frames over the genome. These results significantly enhance our knowledge of RNA virosphere in Blattodea insects, and the novel genome architectures in dicistroviruses and other RNA viruses may break our stereotypes in the understanding of the genomic evolution and the emergence of potential novel viral species.
Dimeric mixed-ligand oxidovanadium complexes V2O2(1,3-pdta)(bpy)2·9H2O (1) and V2O2(1,3-pdta)(phen)2·6H2O (2) feature a symmetric binuclear structure bridged by 1,3-pdta, which is different from ...our previous reported asymmetric binuclear complex V2O2(edta)(phen)2·11H2O (3).In this study, a wide range of analytical techniques were carried out to fully characterize the complexes 1 and 2 and further investigate their structural stabilities. Density functional theory calculations of 1 and 2 also suggest that they might have good reactivity with biomolecules as anticancer agents. To assess and screen the antitumor activities of compounds 1–3 together with their four corresponding monomeric complexes VO(ida)(phen), VO(ida)(bpy), VO(OH)(phen)2Cl, and VO(Hedta)−, we have performed in vitro experiments with hepatocellular carcinoma HepG2 and SMMC-7721 cell lines by MTT analyses. Complex 2 was found to have the highest inhibitory potency against the growth of HepG2 and SMMC-7721 cells (IC50 = 2.07 ± 0.72 μM for HepG2; 13.00 ± 3.06 μM for SMMC-7721) compared to other compounds. The structure–activity relationship studies showed that the antitumor effect of compound 2 is higher than that of other compounds. After studying the monomeric compounds of 1–3, their effects were also ranked. Moreover, complex 2 displayed stronger binding affinity toward calf thymus DNA (K b = 5.71 × 104 M–1) and cleavage activities than the other complexes (K b = 1.34 × 104 M–1 for 1 and 5.22 × 104 M–1 for 3, respectively). We further extended the cellular mechanisms of drug action and found that 2 could block DNA synthesis and cell division of HepG2 and 7721 cells and further induce apoptosis by flow cytometry assays. In short, these results indicate that binuclear oxidovanadium compounds could have potential as simple, effective, and safe antitumor agents.
Herein, we report an engineered enzyme that can monooxygenate unprotected tryptophan into the corresponding 3a‐hydroxyhexahydropyrrolo2,3‐bindole‐2‐carboxylic acid (HPIC) in a single, scalable step ...with excellent turnover number and diastereoselectivity. Taking advantage of directed evolution, we analyzed the stepwise oxygen‐insertion mechanism of tryptophan 2,3‐dioxygenases, and transformed tryptophan 2,3‐dioxygenase from Xanthomonas campestris into a monooxygenase for oxidative cyclization of tryptophans. It was revealed that residue F51 is vital in determining the product ratio of HPIC to N′‐formylkynurenine. Our reactions and purification procedures use no organic solvents, resulting in an eco‐friendly method to prepare HPICs for further applications.
Gerichtete Evolution: Die Tryptophan‐2,3‐Dioxygenase aus Xanthomonas campestris (xcTDO) wurde durch gerichtete Evolution in eine Monooxygenase umgewandelt. Die konstruierte xcTDO weist eine ausgezeichnete Umsatzzahl und hohe Diastereoselektivität auf und ermöglicht die skalierbare Synthese von 3a‐Hydroxyhexahydropyrrolo2,3‐bindol‐2‐carbonsäuren (HPICs) aus Tryptophanen in einem Schritt.
According to the requirement of recognizing traffic police gestures for driver assistance systems and intelligent vehicles, a universal model for dynamic traffic police gesture recognition is firstly ...introduced, of which can accurately present the spatial context (such as the relative lengths of skeletons, the angles between each skeleton w. r. t. gravity, and part features) of the traffic police gestures. Secondly, an architecture which can respectively extract spatial context and temporal features of dynamic traffic police gesture is proposed. Meanwhile, deep neural network and LSTM are introduced to build a high-resolution traffic police gestures recognizer (namely HRTPGR). At last, the open Police Gesture Dataset is used to train and test TPGR, and the experimental results show that the TPGR achieves a state-of-the-art accuracy with 98.7% for dynamic traffic police gestures recognition, and has strong anti-interference ability to light, background and gesture shape changes.
This paper introduces agent-based methodology to build a distributed autonomic storage system infrastructure, and an effectively negotiation mechanism based on agent is applied for data location. We ...present Availability-based Data Allocation （ADA） algorithm as a data placement strategy to achieve high efficient utilization of storage resources by employing multiple distributed storage resources. We use Bloom filter in each storage device to track the location of data. We present the data lookup strategy that small size of read request is handled directly, and large size of read request is handled by cooperation with storage devices.The performance evaluation shows that the data location mechanism is high available and can work well for heterogeneous autonomic storage systems.
（Ti,AI）N films were fabricated by arc ion plating （ALP） and then annealed within a range of temperatures from 200 to 500℃ for 30 rain in vacuum. The results indicate that the average residual ...stresses decrease slightly from -5.84 to -4.98 GPa with increasing annealing temperature. The stress depth distribution evolves from a sharp ＂bell＂ shape to a mild ＂bell＂ shape, suggesting a more uniform stress state in the annealed films. The microstructure of the films was also investigated in detail. The as-deposited film consists of fine columnar crystals with an amorphous phase at the interface. During heat treatment, the columnar subgrain growth was observed; meanwhile, the phenomenon of crystallization has been identified at the interface. Further more, the relationship between the residual stresses and the microstructure of the films was explored and highlighted. In addition, there is no hardness degradation of the films during heat treatment.
