Rational design of antibodies and development of a novel method for (1–3)-β-D glucan detection as an alternative to Limulus amebocyte lysate assay

With advances in medicine, increasing medical interventions have increased the risk of invasive fungal disease development. (1-3)-β-D glucan (BDG) is a common fungal biomarker in serological tests. However, the scarcity of Limulus resources for BDG detection poses a challenge. This study addresses the need for an alternative to Limulus amebocyte lysate by using BDG mutant antibody for chemiluminescence detection. The wild-type BDG antibody was obtained by immunizing rabbits. An optimal V52HI/N34L Y mutant antibody, which has increased 3.7-fold of the testing efficiency compared to the wild-type antibody, was first achieved by mutating “hot-spot” residues that contribute to strong non-covalent bonds, as determined by alanine scanning and molecular dynamics simulation. The mutant was then applied to develop the magnetic particle chemiluminescence method. 574 clinical samples were tested using the developed method, with a cutoff value of 95 pg/mL set by Limulus amebocyte lysate. The receiver operating characteristic curve demonstrated an area under the curve of 0.905 (95% CI: 0.880–0.929). Chemiluminescence detected an antigen concentration of 89.98 pg/mL, exhibiting a sensitivity of 83.33% and specificity of 89.76%. In conclusion, the results showed a good agreement with Limulus amebocyte lysate and demonstrated the feasibility of using BDG mutant antibodies for invasive fungal disease diagnosis. The new method based on chemiluminescence for detecting BDG could shorten the sample-to-result time to approximately 30 min, rescue Limulus from being endangered and is resource efficient in terms of equipment and the non-use of a skilled technician

Cellulase Recycling after High-Solids Simultaneous Saccharification and Fermentation of Combined Pretreated Corncob

Despite the advantageous prospect of second-generation bioethanol, its final commercialization must overcome the primary cost impediment due to enzyme assumption. To solve this problem, this work achieves high-concentration ethanol fermentation and multi-round cellulase recycling through process integration. The optimal time and temperature of the re-adsorption process were determined by monitoring the adsorption kinetics of cellulases. Both glucose and cellobiose inhibited cellulase adsorption. After 96 h of ethanol fermentation, 40% of the initial cellulase remained in the broth, from which 62.5% of the cellulase can be recycled and reused in fresh substrate re-adsorption for 90 min. Under optimum conditions, i.e., pH 5.0, dry matter loading of 15 wt%, cellulase loading of 45 FPU/g glucan, two cycles of fermentation and re-adsorption can yield two-fold increased ethanol outputs and reduce enzyme costs by over 50%. The ethanol concentration in each cycle can be achieved at levels greater than 40 g/L

Poly[penta­kis­(μ-cyanido-κ2 N:C)tris­(5-phenyl-2,2′-bipyridine-κ2 N,N′)penta­copper(I)]

The hydro­thermal reaction of Cu(acetate)2 and K3[Fe(CN)6] with 5-phenyl-2,2′-bipyridine (5-ph-2,2′-bpy) in water yields the polymeric title complex, [Cu5(CN)5(C16H12N2)3]n, which consists of ribbons along the a axis, constructed from 26-membered {Cu10(CN)8} rings. In these rings, the metal atoms are bridged by cyanide groups, except for one close Cu⋯Cu contact [2.7535 (12) Å], which can be considered as ligand-unsupported. Within the rings, one Cu atom has a distorted tetra­hedral geometry through the coordination to two N atoms from 5-ph-2,2′-bpy and two N/C atoms from two cyanide groups. Two Cu atoms have a trigonal planar environment being coordinated by three cyanide groups and two other Cu atoms have a distorted square planar geometry through coordination to two N atoms from 5-ph-2,2′-bpy and two N/C atoms from two cyanide groups

Gold Nanoparticle-Aptamer-Based LSPR Sensing of Ochratoxin A at a Widened Detection Range by Double Calibration Curve Method

Ochratoxin A (OTA) is a type of mycotoxin generated from the metabolism of Aspergillus and Penicillium, and is extremely toxic to humans, livestock, and poultry. However, traditional assays for the detection of OTA are expensive and complicated. Other than OTA aptamer, OTA itself at high concentration can also adsorb on the surface of gold nanoparticles (AuNPs), and further inhibit AuNPs salt aggregation. We herein report a new OTA assay by applying the localized surface plasmon resonance effect of AuNPs and their aggregates. The result obtained from only one single linear calibration curve is not reliable, and so we developed a “double calibration curve” method to address this issue and widen the OTA detection range. A number of other analytes were also examined, and the structural properties of analytes that bind with the AuNPs were further discussed. We found that various considerations must be taken into account in the detection of these analytes when applying AuNP aggregation-based methods due to their different binding strengths

Modeling and Testing of a Composite Steel–Concrete Joint for Hybrid Girder Bridges

A hybrid girder bridge adopts a steel segment at the mid-span of the main span of a continuous concrete girder bridge. The critical point of the hybrid solution is the transition zone, connecting the steel and concrete segments of the beam. Although many girder tests revealing the structural behavior of hybrid girders have been conducted by previous studies, few specimens took the full section of a steel–concrete joint due to the large size of prototype hybrid bridges. In this study, a static load test on a composite segment to bridge the joint between the concrete and steel parts of a hybrid bridge with full section was conducted. A finite element model replicating the tested specimen results was established through Abaqus, while parametric studies were also conducted. The test and numerical results revealed that the concrete filling in the composite solution prevented the steel flange from extensive buckling, which significantly improved the load-carrying capacity of the steel–concrete joint. Meanwhile, strengthening the interaction between the steel and concrete helps to prevent the interlayer slip and simultaneously contributes to a higher flexural stiffness. These results are an important basis for establishing a rational design scheme for the steel–concrete joint of hybrid girder bridges

Identification of hub genes and therapeutic drugs in osteonecrosis of the femoral head through integrated bioinformatics analysis and literature mining

Abstract Osteonecrosis of the femoral head (ONFH) is a multifactorial disease leading to severely limited function. By far, the etiology and pathogenesis of ONFH are not fully understood, and surgery is the only effective way to treat ONFH. This study aims to identify hub genes and therapeutic drugs in ONFH. Two gene expression profiles were downloaded from the gene expression omnibus database, and the hub genes and candidate drugs for ONFH were identified through integrated bioinformatics analysis and cross-validated by literature mining. A total of 159 DEGs were identified. PTGS2, LRRK2, ANXA5, IGF1R, MCL1, TIMP2, LYN, CD68, CBL, and RUNX2 were validated as 10 hub genes, which has considerable implications for future genetic research and related research fields of ONFH. Our findings indicate that 85 drugs interact with ONFH, with most drugs exhibiting a positive impact on ONFH by promoting osteogenesis and angiogenesis or inhibiting microcirculation embolism, rather than being anti-inflammatory. Our study provides novel insights into the pathogenesis, prevention, and treatment of ONFH

Effect of hydrophobicity and charge separation on the antifouling properties of surface-tethered zwitterionic peptides

Zwitterionic peptides emerge as a class of highly effective antifouling materials in a wide range of applications such as biosensor, biomedical devices, and implants. We incorporated neutral amino acid spacers with different hydrophobicities, including serine (Ser), glycine (Gly), and leucine (Leu), into zwitterionic peptides with KE repeating units and investigated the structure and antifouling performance of the zwitterionic peptide brushes using surface plasma resonance (SPR), surface force apparatus (SFA), and all atomistic molecular dynamics (MD) simulation techniques. Our results demonstrate that the hydrophilicity of neutral spacers alters the structure and antifouling performance of the peptide-modified surface. Hydrophilic Ser inserted peptides reduced the interaction between the peptide monolayer and protein foulants while hydrophobic Leu significantly increased the protein adhesion. SFA force measurements shows that the presence of more spacers would increase the adhesion between the peptide monolayer and the modeling foulant lysozyme, especially for the hydrophobic spacers. MD simulations reveal that hydrophilic Ser spacers retain the hydrophilicity of the peptide monolayer and improve the antifouling performance, and Gly spacers give rise to more inter-chain crosslinks. Leu spacers result in a more hydrophobic peptide monolayer which lead to dehydration of the peptide monolayer and reduces the antifouling performances.Ministry of Education (MOE)National Research Foundation (NRF)Accepted versionC.L. and R.S. acknowledge the National Natural Science Foundation of China (21621004), the Tianjin Municipal Science and Technology Bureau, China (16JCZDJC37900), the Ministry of Education (grant no. NCET-11-0372), and financial support from the China Scholarship Council (CSC, 201806250100). M.L. and J.Y. acknowledge the Singapore Ministry of Education Academic Research Fund Tier 1 (RG7/ 19) and the Singapore National Research Fellowship (NRFNRFF11-2019-0004)