Fatigue Strength Analysis of Bogie Frame In Consideration of Parameter Uncertainty

In this paper, a fatigue strength analysis approach based on Goodman-Smith fatigue limit diagram （GSFLD） and reliability theory is proposed to solve the problem that the traditional fatigue strength analysis of bogie frame is too conservative, considering the parameter uncertainty in engineering practice. Firstly, according to UIC615-4, EN13749 standard and GSFLD, the fatigue strength of the frame is calculated. The experimental results are compared with the simulation data to determine the location of the higher fatigue strength as the control point for the strength evaluation. Secondly, the parametric model of the frame is established by APDL language, and the D-optimal experiment design of uncertainty parameters is carried out. The polynomial response surface function with the mean stress and stress amplitude of the control point as the objective is established. The control points under the influence of uncertainty parameters are obtained by importance sampling method. Finally, the functional expression of GSFLD without considering the safety factor is derived, and the fatigue strength reliability of control point is calculated. The results of this study not only reveal the influence of parameter uncertainty on fatigue strength, but also demonstrate a need of developing new evaluation methods to accommodate fatigue analysis

Fatigue strength analysis of bogie frame in consideration of parameter uncertainty

In this paper, a fatigue strength analysis approach based on Goodman-Smith fatigue limit diagram (GSFLD) and reliability theory is proposed to solve the problem that the traditional fatigue strength analysis of bogie frame is too conservative, considering the parameter uncertainty in engineering practice. Firstly, according to UIC615-4, EN13749 standard and GSFLD, the fatigue strength of the frame is calculated. The experimental results are compared with the simulation data to determine the location of the maximum fatigue strength as the control point for the strength evaluation. Secondly, the parametric model of the frame is established by ANSYS parametric design language (APDL) language, and the D-optimal experiment design of uncertainty parameters is carried out. The polynomial response surface function with the mean stress and stress amplitude of the control point as the objective is established. The control points under the influence of uncertainty parameters are obtained by importance sampling method. Finally, the functional expression of GSFLD without considering the safety factor is derived, and the fatigue strength reliability of control point is calculated. The results of this study not only reveal the influence of parameter uncertainty on fatigue strength, but also demonstrate a need of developing new evaluation methods to accommodate fatigue analysis

Application of a biofilm-enhanced A2O system in the treatment of wastewater from mariculture

Development of environment-friendly and efficient aquaculture effluent treatment system is crucial for sustainable intensification of aquaculture, in the face of the rapidly increasing environmental pressure in the mariculture industry. In this study, mariculture wastewater was treated by the anoxic-anaerobic-oxic biochemical treatment system (A2O system) with traditional activated sludge replaced by nitrifying bacteria, denitrification bacteria and phosphorus accumulating bacteria absorbed on PBS carrier biofilms suitable for saline/brackish water. The results showed that biofilm-enhanced A2O system can effectively remove pollutants from aquaculture wastewater. The removal efficiencies of CODMn, NH4+-N, TN and TP in A2O system were approximately 86.3%-90.8%, 97.7%-99.5%, 94.6%-95.2% and 97.0%-98.1%. The results further showed that CODMn, NH4+-N, and TN were mainly removed in anaerobic tank and anoxic tank, while TP was mainly removed in the anoxic tank and oxic tank. The biofilm-enhanced A2O system by adding nitrifying bacteria and phosphorus accumulating bacteria biofilms using PBS as carriers instead of conventional activated sludge could be applied to the treatment of circulating aquaculture wastewater. This study provides a feasible scheme for enhancing the efficiency of A2O system in the treatment of aquaculture tail water, and provides a reference for the immobilization of microorganisms

A practical guide to promote informatics-driven efficient biotopographic material development

Micro/nano topographic structures have shown great utility in many biomedical areas including cell therapies, tissue engineering, and implantable devices. Computer-assisted informatics methods hold great promise for the design of topographic structures with targeted properties for a specific medical application. To benefit from these methods, researchers and engineers require a highly reusable “one structural parameter – one set of cell responses” database. However, existing confounding factors in topographic cell culture devices seriously impede the acquisition of this kind of data. Through carefully dissecting the confounding factors and their possible reasons for emergence, we developed corresponding guideline requirements for topographic cell culture device development to remove or control the influence of such factors. Based on these requirements, we then suggested potential strategies to meet them. In this work, we also experimentally demonstrated a topographic cell culture device with controlled confounding factors based on these guideline requirements and corresponding strategies. A “guideline for the development of topographic cell culture devices” was summarized to instruct researchers to develop topographic cell culture devices with the confounding factors removed or well controlled. This guideline aims to promote the establishment of a highly reusable “one structural parameter – one set of cell responses” database that could facilitate the application of informatics methods, such as artificial intelligence, in the rational design of future biotopographic structures with high efficacy

A β-glucosidase hyper-production Trichoderma reesei mutant reveals a potential role of cel3D in cellulase production

Abstract Background The conversion of cellulose by cellulase to fermentable sugars for biomass-based products such as cellulosic biofuels, biobased fine chemicals and medicines is an environment-friendly and sustainable process, making wastes profitable and bringing economic benefits. Trichoderma reesei is the well-known major workhorse for cellulase production in industry, but the low β-glucosidase activity in T. reesei cellulase leads to inefficiency in biomass degradation and limits its industrial application. Thus, there are ongoing interests in research to develop methods to overcome this insufficiency. Moreover, although β-glucosidases have been demonstrated to influence cellulase production and participate in the regulation of cellulase production, the underlying mechanism remains unclear. Results The T. reesei recombinant strain TRB1 was constructed from T. reesei RUT-C30 by the T-DNA-based mutagenesis. Compared to RUT-C30, TRB1 displays a significant enhancement of extracellular β-glucosidase (BGL1) activity with 17-fold increase, a moderate increase of both the endoglucanase (EG) activity and the exoglucanase (CBH) activity, a minor improvement of the total filter paper activity, and a faster cellulase induction. This superiority of TRB1 over RUT-C30 is independent on carbon sources and improves the saccharification ability of TRB1 cellulase on pretreated corn stover. Furthermore, TRB1 shows better resistance to carbon catabolite repression than RUT-C30. Secretome characterization of TRB1 shows that the amount of CBH, EG and BGL in the supernatant of T. reesei TRB1 was indeed increased along with the enhanced activities of these three enzymes. Surprisingly, qRT-PCR and gene cloning showed that in TRB1 β-glucosidase cel3D was mutated through the random insertion by AMT and was not expressed. Conclusions The T. reesei recombinant strain TRB1 constructed in this study is more desirable for industrial application than the parental strain RUT-C30, showing extracellular β-glucosidase hyper production, high cellulase production within a shorter time and a better resistance to carbon catabolite repression. Disruption of β-glucosidase cel3D in TRB1 was identified, which might contribute to the superiority of TRB1 over RUT-C30 and might play a role in the cellulase production. These results laid a foundation for future investigations to further improve cellulase enzymatic efficiency and reduce cost for T. reesei cellulase production.http://deepblue.lib.umich.edu/bitstream/2027.42/134636/1/12934_2016_Article_550.pd

Preventive Effect of Volvariella volvacea Fruit Body Polypeptides on Acute Alcoholic Liver Injury in Mice and Its Influence on Intestinal Microflora

To investigate the preventive effect of Volvariella volvacea fruit body polypeptides (VVFP) on acute alcoholic liver injury in mice and its influence on the intestinal microbiota, VVFP (1–3 kDa molecular mass) which had been previously obtained by our laboratory was given by gavage to mice. The mice were randomly divided into six groups: blank control, model, positive control, low-dose VVFP, moderate-dose VVFP and high-dose VVFP. Serum indexes, liver indexes and histopathological sections were compared among these groups, and 16S rDNA gene high-throughput sequencing was used to analyze the diversity of the intestinal microflora and the relative abundance at the phyla and genus levels in each sample. Results showed that VVFP significantly reduced the levels of triglycerides (TG), total cholesterol (TC), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activity in the serum and malondialdehyde (MDA) in the liver, and decreased the levels of the inflammatory cytokines tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6), and significantly increased the activities of alcohol dehydrogenase (ADH), total superoxide dismutase (T-SOD) and glutathione peroxidase (GSH-PX) in the liver. The 16S rRNA sequencing showed that VVFP significantly reduced the α-diversity indices Chao1 and observed species, increased the Shannon index, and regulated the abundance of Bacteroides, Firmicutes, Streptomyces, Lactobacillus and Vibrio, thereby reducing liver damage. In conclusion, VVFP can reduce alcoholic liver injury, which will provide a theoretical basis for the application of VVFP in the field of functional foods

Twelve-year outcomes of bedside laser photocoagulation for severe retinopathy of prematurity

PurposeThe purpose of this study is to evaluate the 12-year outcomes of bedside laser photocoagulation (LP) for severe retinopathy of prematurity (ROP) under sedation combined with ocular surface anesthesia in neonatal intensive care units (NICU).DesignThe study is a retrospective case series.MethodsInfants treated with bedside LP for severe ROP from April 2009 to September 2021 were included. All LP treatments were performed under sedation and surface anesthesia at the bedside in NICU. Data were recorded for clinical and demographic characteristics, total laser spots, duration of treatment, proportion of total regression of ROP, proportion of recurrence, and adverse events.ResultsA total of 364 infants (715 eyes) were included, with a mean gestational age of 28.6 ± 2.4 weeks (range: 22.6–36.6 weeks) and a mean birth weight of 1,156.0 ± 339.0 g (range: 480–2,200 g). The mean number of laser spots was 832 ± 469, and the mean duration of treatment was 23.5 ± 5.3 min per eye. Of all the eyes, 98.3% responded to LP with complete regression of ROP. ROP recurred in 15 (2.1%) eyes after the initial LP. Additional LP was performed in seven (1.0%) eyes. No patient exhibited mistaken LP of other ocular tissues, and there were no serious ocular adverse effects. None of them needed endotracheal intubation.ConclusionsBedside LP treatment is effective and safe for premature infants with severe ROP under sedation and surface anesthesia in NICU, especially for infants whose general condition is unstable and not suitable for transport

Deep functional analysis of synII, a 770-kilobase synthetic yeast chromosome

INTRODUCTION Although much effort has been devoted to studying yeast in the past few decades, our understanding of this model organism is still limited. Rapidly developing DNA synthesis techniques have made a “build-to-understand” approach feasible to reengineer on the genome scale. Here, we report on the completion of a 770-kilobase synthetic yeast chromosome II (synII). SynII was characterized using extensive Trans-Omics tests. Despite considerable sequence alterations, synII is virtually indistinguishable from wild type. However, an up-regulation of translational machinery was observed and can be reversed by restoring the transfer RNA (tRNA) gene copy number. RATIONALE Following the “design-build-test-debug” working loop, synII was successfully designed and constructed in vivo. Extensive Trans-Omics tests were conducted, including phenomics, transcriptomics, proteomics, metabolomics, chromosome segregation, and replication analyses. By both complementation assays and SCRaMbLE (synthetic chromosome rearrangement and modification by loxP -mediated evolution), we targeted and debugged the origin of a growth defect at 37°C in glycerol medium. RESULTS To efficiently construct megabase-long chromosomes, we developed an I- Sce I–mediated strategy, which enables parallel integration of synthetic chromosome arms and reduced the overall integration time by 50% for synII. An I- Sce I site is introduced for generating a double-strand break to promote targeted homologous recombination during mitotic growth. Despite hundreds of modifications introduced, there are still regions sharing substantial sequence similarity that might lead to undesirable meiotic recombinations when intercrossing the two semisynthetic chromosome arm strains. Induction of the I- Sce I–mediated double-strand break is otherwise lethal and thus introduced a strong selective pressure for targeted homologous recombination. Since our strategy is designed to generate a markerless synII and leave the URA3 marker on the wild-type chromosome, we observed a tenfold increase in URA3 -deficient colonies upon I- Sce I induction, meaning that our strategy can greatly bias the crossover events toward the designated regions. By incorporating comprehensive phenotyping approaches at multiple levels, we demonstrated that synII was capable of powering the growth of yeast indistinguishably from wild-type cells (see the figure), showing highly consistent biological processes comparable to the native strain. Meanwhile, we also noticed modest but potentially significant up-regulation of the translational machinery. The main alteration underlying this change in expression is the deletion of 13 tRNA genes. A growth defect was observed in one very specific condition—high temperature (37°C) in medium with glycerol as a carbon source—where colony size was reduced significantly. We targeted and debugged this defect by two distinct approaches. The first approach involved phenotype screening of all intermediate strains followed by a complementation assay with wild-type sequences in the synthetic strain. By doing so, we identified a modification resulting from PCRTag recoding in TSC10 , which is involved in regulation of the yeast high-osmolarity glycerol (HOG) response pathway. After replacement with wild-type TSC10 , the defect was greatly mitigated. The other approach, debugging by SCRaMbLE, showed rearrangements in regions containing HOG regulation genes. Both approaches indicated that the defect is related to HOG response dysregulation. Thus, the phenotypic defect can be pinpointed and debugged through multiple alternative routes in the complex cellular interactome network. CONCLUSION We have demonstrated that synII segregates, replicates, and functions in a highly similar fashion compared with its wild-type counterpart. Furthermore, we believe that the iterative “design-build-test-debug” cycle methodology, established here, will facilitate progression of the Sc2.0 project in the face of the increasing synthetic genome complexity. SynII characterization. ( A ) Cell cycle comparison between synII and BY4741 revealed by the percentage of cells with separated CEN2-GFP dots, metaphase spindles, and anaphase spindles. ( B ) Replication profiling of synII (red) and BY4741 (black) expressed as relative copy number by deep sequencing. ( C ) RNA sequencing analysis revealed that the significant up-regulation of translational machinery in synII is induced by the deletion of tRNA genes in synII. </jats:sec