β-1,3-Glucan-Induced Host Phospholipase D Activation Is Involved in Aspergillus fumigatus Internalization into Type II Human Pneumocyte A549 Cells

The internalization of Aspergillus fumigatus into lung epithelial cells is a process that depends on host cell actin dynamics. The host membrane phosphatidylcholine cleavage driven by phospholipase D (PLD) is closely related to cellular actin dynamics. However, little is known about the impact of PLD on A. fumigatus internalization into lung epithelial cells. Here, we report that once germinated, A. fumigatus conidia were able to stimulate host PLD activity and internalize more efficiently in A549 cells without altering PLD expression. The internalization of A. fumigatus in A549 cells was suppressed by the downregulation of host cell PLD using chemical inhibitors or siRNA interference. The heat-killed swollen conidia, but not the resting conidia, were able to activate host PLD. Further, β-1,3-glucan, the core component of the conidial cell wall, stimulated host PLD activity. This PLD activation and conidia internalization were inhibited by anti-dectin-1 antibody. Indeed, dectin-1, a β-1,3-glucan receptor, was expressed in A549 cells, and its expression profile was not altered by conidial stimulation. Finally, host cell PLD1 and PLD2 accompanied A. fumigatus conidia during internalization. Our data indicate that host cell PLD activity induced by β-1,3-glucan on the surface of germinated conidia is important for the efficient internalization of A. fumigatus into A549 lung epithelial cells

Brachial-ankle pulse wave velocity as a risk factor for high body fat mediated by blood pressure

Background Brachial-ankle pulse wave velocity (baPWV) is an important clinical indicator of aortic stiffness and a risk predictor of cardiovascular disease and associated with obesity. However, whether body mass index (BMI) is associated with baPWV remains controversial. In our study, body fat-related indicators, including BMI, body fat rate (BFR), body fat volume (BFV), waist circumference (WC) were examined from healthy volunteers. We investigated the correlation of baPWV with these indicators and also assessed whether baPWV has the potential to predict these indicators. Methods A total of 429 healthy participants were enrolled in this study. Body fat indices, blood pressures, baPWV and blood metabolic indices were measured and recorded. The association of baPWV and indices reflecting body fat and blood pressure, as well as mediation effect were analyzed. Results Three different types of baPWV values were significantly correlated. Mean level of baPWV was an independent risk factor for WC, BMI, BFR, and BFV (exp(β) = 1.011, 1.004, 1.010 and 1.009, respectively, P < .001 for all) but not BMR. As for mediation effects, baPWV positively influenced WC (Total effect = 0.011, P < .001), BMI (Total effect = 0.004, P < .001) and BFV (Total effect = 0.009, P < .001) in indirect way mediated by SBP and DBP, while baPWV influenced BFR in both direct (Effect = 0.004, P = .018) and indirect way. Conclusions Levels of baPWV correlated with obesity and is an independent risk factor for WC, BMI, BFR and BFV. Besides, baPWV positively associated with WC, BMI and BFV mainly in indirect way mediated by SBP and DBP, and baPWV associated with BFR in both direct and indirect way

Effectiveness of PIVKA-II in the detection of hepatocellular carcinoma based on real-world clinical data

Abstract Background Protein Induced by Vitamin K Absence or Antagonist-II (PIVKA-II) is an efficient biomarker specific for hepatocellular carcinoma (HCC). Some researchers have proved that levels of PIVKA-II reflect HCC oncogenesis and progression. However, the effectiveness of PIVKA-II based on real-world clnical data has barely been studied. Methods A total of 14,861 samples were tested in Southwest Hospital in over 2 years’ time. Among them, 4073 samples were PIVKA-II positive. Finally, a total of 2070 patients with at least two image examinations were enrolled in this study. Levels of AFP and PIVKA-II were measured by chemiluminescence enzyme immunoassay (CLEIA) and chemiluminescent microparticle Immunoassay (CMIA), respectively. Results A total of 1016 patients with HCC were detected by PIVKA-II in a real-world application. In all these cases, 88.7% cases primarily occurred and patients with advanced HCC covered 61.3%. Levels of PIVKA-II were significantly higher in advanced group (4650.0 mAU/ml, 667.0–33,438.0 mAU/ml) than early-stage group (104.5 mAU/ml, 61.0–348.8 mAU/ml; P < 0.001). Levels of PIVKA-II elevated significantly in recurrence and residual group than recovery group (P < 0.001). A total of 1054 PIVKA-II positive patients were non-HCC cases. Among them, cirrhosis took the largest part (46.3%), followed by hepatitis (20.6%) and benign nodules (15.3%). High-levels of PIVKA-II in at-risk patients is an indicator of HCC development in two-year time. Conclusions Our data showed that PIVKA-II effectively increases the detection rate of HCC was a valid complement to AFP and image examination in HCC surveillance

A novel mutation in the BCHE gene and phenotype identified in a child with low butyrylcholinesterase activity: a case report

Abstract Background Butyrylcholinesterase (BChE), an ester hydrolase produced mainly by the liver, hydrolyzes certain short-acting neuromuscular blocking agents, like succinylcholine and mivacurium that are widely used during anesthesia. Patients with BChE deficiency are possibly in danger of postanesthetic apnea. Hereditary BChE deficiency results from the mutations of BCHE gene located on chromosome 3, 3q26.1-q26.2, between nucleotides 165,490,692–165,555,260. Case presentation This study describes a novel mutation in a child with BChE deficiency. In general, this child appeared healthy and well-developed with a normal appearance. However, the results of Wechsler Intelligence Scale showed that the full-scale intelligence quotient (FIQ) was 53, classified into the group with the minor defect. The BChE activity was 32.0 U/L, considerably lower than the normal lower limit (reference range: 5000-12,000 U/L). Sanger sequencing showed that there were 2 mutations in the exon 2 of BCHE gene of this child. One is a heterozygous mutation rs764588882 (NM_000055.3: c.401_402insA, p.Asn134Lysfs*23). The other one is a heterozygous mutation (NM_000055.3: c.73A > T, p.Lys25Ter) that has never been reported before. The two mutations lead to a premature stop of transcription. Conclusions Double heterozygous recessive mutations are the cause of BChE deficiency of this boy in this study, including a novel mutation c.73A > T. Intellectual disability is a new phenotype that is probably associated with this mutation

Probabilistic-assured resource provisioning with customizable hybrid isolation for vertical industrial slicing

Abstract With the increasing demand of network slices in vertical industries, slice resource provisioning in transport networks has encountered two challenges, one is efficient slice resource provisioning in the presence of traffic uncertainty of slices, and another is flexible slice resource isolation for customizable isolation needs. In this paper, we propose an innovative flexible hybrid isolation model to support any customized resource isolation from complete isolation to full sharing, and solve the slice resource provisioning problem named Hybrid Slicing Minimum Bandwidth (HSMB) by considering traffic prediction error to mitigate the negative impact of traffic uncertainty in the proposed model. After analyzing the HSMB problem, 1) we first try to solve the problem in steps and decompose the HSMB problem into grouping sub-problem and adjusting sub-problem, 2) we then propose a low-complexity dynamic programming grouping algorithm and a fast iterative adjustment algorithm for the two sub-problems based on probabilistic feature-based analysis, 3) we combine the algorithms of the two sub-problems and further propose a linking algorithm for the potential insufficient resource dilemma and high computational complexity dilemma to improve the efficiency of the solution. The numerical results show that the proposed flexible hybrid isolation model with different factors can facilitate flexible slice isolation with customized isolation demands, while the proposed algorithm can realize efficient slice resource provisioning with a probabilistic guarantee. The comparison result shows the proposed algorithms outperform the other benchmark algorithms

Table_1_Ae1/Sbe1 maize-derived high amylose improves gut barrier function and ameliorates type II diabetes in high-fat diet-fed mice by increasing Akkermansia.docx

Type II diabetes mellitus (T2DM) has its origins in chronic inflammation due to immune dysregulation. Improving chronic inflammation can significantly reduce the probability of T2DM and the rate of disease progression. Resistance to starch 2 (RSII) high-amylose maize starch (HAMS) has been widely implicated in the improvement and regulation of T2DM. However, its exact molecular mechanisms have not been fully discovered. Here, we used CRISPR/Cas9 technology to knock out two starch-branching enzyme genes, Ae1 and Sbe1, in maize to obtain mutants containing higher levels of HAMS. In experiments in which HAMS was fed to mice on a high-fat diet (HFD), we confirmed the function of HAMS in ameliorating hyperglycemia. Mechanistically, we found that HAMS improves the gut barrier function by increasing the Akkermansia abundance in the gut. This increase led to the alleviation of chronic inflammation in mice on a HFD, resulting in improved insulin sensitivity and a decrease in blood glucose.</p

Table_2_Ae1/Sbe1 maize-derived high amylose improves gut barrier function and ameliorates type II diabetes in high-fat diet-fed mice by increasing Akkermansia.docx

Type II diabetes mellitus (T2DM) has its origins in chronic inflammation due to immune dysregulation. Improving chronic inflammation can significantly reduce the probability of T2DM and the rate of disease progression. Resistance to starch 2 (RSII) high-amylose maize starch (HAMS) has been widely implicated in the improvement and regulation of T2DM. However, its exact molecular mechanisms have not been fully discovered. Here, we used CRISPR/Cas9 technology to knock out two starch-branching enzyme genes, Ae1 and Sbe1, in maize to obtain mutants containing higher levels of HAMS. In experiments in which HAMS was fed to mice on a high-fat diet (HFD), we confirmed the function of HAMS in ameliorating hyperglycemia. Mechanistically, we found that HAMS improves the gut barrier function by increasing the Akkermansia abundance in the gut. This increase led to the alleviation of chronic inflammation in mice on a HFD, resulting in improved insulin sensitivity and a decrease in blood glucose.</p

Inhibition of <i>A. fumigatus</i> internalization by PLD chemical inhibitors.

<p>A549 cells were incubated for 30 min with 1% (v/v) 1-butanol or tert-butanol (A, B), 2 nM VU0359595 (PLD1-specific inhibitor), 100 nM VU0285655-1 (PLD2-specific inhibitor), or both (C, D). Subsequently, the cells were infected with <i>A. fumigatus</i> 13073 swollen conidia at an MOI of 10. <i>A. fumigatus</i> internalization was analyzed by the nystatin protection assay (A, C) and PLD activities (B, D) were measured. Differences in [³H] PtdEtOH formation or <i>A. fumigatus</i> internalization between the untreated (control) cells and inhibitor-pretreated cells were compared. Data are represented as the mean ± SE (n = 3–4). *P<0.05.</p

Disruption of the Phospholipase D Gene Attenuates the Virulence of Aspergillus fumigatus

Aspergillus fumigatus is the most prevalent airborne fungal pathogen that induces serious infections in immunocompromised patients. Phospholipases are key enzymes in pathogenic fungi that cleave host phospholipids, resulting in membrane destabilization and host cell penetration. However, knowledge of the impact of phospholipases on A. fumigatus virulence is rather limited. In this study, disruption of the pld gene encoding phospholipase D (PLD), an important member of the phospholipase protein family in A. fumigatus, was confirmed to significantly decrease both intracellular and extracellular PLD activity of A. fumigatus. The pld gene disruption did not alter conidial morphological characteristics, germination, growth, and biofilm formation but significantly suppressed the internalization of A. fumigatus into A549 epithelial cells without affecting conidial adhesion to epithelial cells. Importantly, the suppressed internalization was fully rescued in the presence of 100 μM phosphatidic acid, the PLD product. Indeed, complementation of pld restored the PLD activity and internalization capacity of A. fumigatus. Phagocytosis of A. fumigatus conidia by J774 macrophages was not affected by the absence of the pld gene. Pretreatment of conidia with 1-butanol and a specific PLD inhibitor decreased the internalization of A. fumigatus into A549 epithelial cells but had no effect on phagocytosis by J774 macrophages. Finally, loss of the pld gene attenuated the virulence of A. fumigatus in mice immunosuppressed with hydrocortisone acetate but not with cyclophosphamide. These data suggest that PLD of A. fumigatus regulates its internalization into lung epithelial cells and may represent an important virulence factor for A. fumigatus infection