Platelet activation and aggregation underlie acute thrombosis that leads to ST-elevation myocardial infarction (STEMI). L5—highly electronegative low-density lipoprotein (LDL)—is significantly ...elevated in patients with STEMI. Thus, we examined the role of L5 in thrombogenesis. Plasma LDL from patients with STEMI (n = 30) was chromatographically resolved into 5 subfractions (L1-L5) with increasing electronegativity. In vitro, L5 enhanced adenosine diphosphate–stimulated platelet aggregation twofold more than did L1 and induced platelet-endothelial cell (EC) adhesion. L5 also increased P-selectin expression and glycoprotein (GP)IIb/IIIa activation and decreased cyclic adenosine monophosphate levels (n = 6, P < .01) in platelets. In vivo, injection of L5 (5 mg/kg) into C57BL/6 mice twice weekly for 6 weeks shortened tail bleeding time by 43% (n = 3; P < .01 vs L1-injected mice) and increased P-selectin expression and GPIIb/IIIa activation in platelets. Pharmacologic blockade experiments revealed that L5 signals through platelet-activating factor receptor and lectin-like oxidized LDL receptor-1 to attenuate Akt activation and trigger granule release and GPIIb/IIIa activation via protein kinase C-α. L5 but not L1 induced tissue factor and P-selectin expression in human aortic ECs (P < .01), thereby triggering platelet activation and aggregation with activated ECs. These findings indicate that elevated plasma levels of L5 may promote thrombosis that leads to STEMI.
•Highly electronegative LDL (L5), which is elevated in patients with STEMI, induces platelet activation and aggregation through LOX-1.•L5 may have a role in promoting thrombogenesis that leads to STEMI.
This study presents a proof-of-concept data reduction and analysis protocol that can be applied to the study of polluted sediments. Sediment cores from the Nankan River estuary are used as an example ...of how the protocol can be employed to quantify temporal heavy metal variability. The measurement protocol produces more detailed elemental profiles than conventional techniques using a combination of data transformation techniques and multivariate analysis. Conventional sediment analyses are used to confirm the robustness of the protocol by comparisons of heavy metal concentrations.
X-ray fluorescence (XRF) core scanning provides rapid, high-resolution elemental profiles from sediment cores. The technique relies on a variety of calibration methods (ratio, additive and centred log-ratio) to transform the raw data and reduce bias caused by matrix and closed-sum effects. We further test all these calibration approaches since the transformation process is an essential step for the follow up multivariate analyses. The combination of principal component and cluster analysis objectively assesses the information implicit in the dataset. The settings in each procedure are optimized to account for the variance of the dataset. This optimization protocol explains the heavy metal trends using the sediment characteristics of the cores. Heavy metal pollution is characterized by three periods and classified by their oxidation states. We show that heavy metals have an affinity with fine-grained sediments and Mn. The interpretation is confirmed by grain size analysis and inductively coupled plasma optical emission spectrometry (ICP-OES) measurements. This study provides an impartial, cost- and time-effective protocol suitable for the analysis of other heavy metal polluted sites and further studies using sediment cores as archives.
L5, a highly electronegative subtype of low-density lipoprotein (LDL), is likely associated with the development of atherosclerosis and cardiovascular diseases. Normal LDL is composed mainly of ...apolipoprotein (Apo) B, but L5 has additional proteins such as ApoE. We previously demonstrated that L5 induces endothelial cell senescence by increasing mitochondrial reactive oxygen species. In the present study, we examined the effect of L5 on mitochondrial function in cardiomyocytes.
We used the Seahorse XF24 extracellular flux analyzer to examine the effect of L5 and its components on mitochondrial energy production. The effects of L5 on mitochondrial morphology were examined by immunofluorescence using MitoTracker Green FM and the corresponding probes in H9c2 cardiomyoblasts. Mitochondrial permeability was assessed by using a calcium-induced swelling assay with a voltage-dependent anion-selective channel (VDAC) inhibitor to determine VDAC-dependence both in vitro and in vivo. L5 without ApoE, referred to as △L5, was used to clarify the role of ApoE in L5-induced mitochondrial dysfunction.
L5 not only significantly decreased basal (P < 0.05) and maximal respiration (P < 0.01) but also reduced spare respiratory capacity (P < 0.01) in H9c2 cells. Additionally, L5 caused phosphorylation of Drp1 and mitochondrial fission. Recombinant ApoE mimicked the mitochondrial effects of L5, but △L5 did not cause similar effects. After entering cells, ApoE on L5 colocalized with mitochondrial VDAC and caused mitochondria swelling both in vitro and in vivo. This effect was also seen with recombinant ApoE but not △L5.
ApoE may play an important role in electronegative LDL-induced mitochondrial dysfunction through the opening of the mitochondrial permeability transition pore via the interaction of ApoE and VDAC.
•Electronegative LDL (L5) induced dysfunction of mitochondrial energy production.•ApoE was released and colocalized with VDAC after L5 was taken up by myocytes.•Recombinant ApoE mimicked the effect of L5 on cardiomyocytes.
Sepsis, which is a systemic inflammatory response that follows a bacterial infection, has a high mortality rate and limited therapeutic options. Here we show that the antimicrobial peptide OH-CATH30, ...which naturally occurs in snake, selectively regulates the innate immune response to protect mice from lethal sepsis. The administration of OH-CATH30 significantly improves the survival rate of mice infected by antibiotic-susceptible and -resistant pathogens, including Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus. OH-CATH30 selectively up-regulates the production of chemokines and cytokines without harmful immune response. Recruitment of monocytes, macrophages, and neutrophils to the infection site is pivotal to the protective capacity of OH-CATH30. Furthermore, the alternative activation of the innate immune response by OH-CATH30 depends on p38 mitogen-activated protein kinase signaling. Taken together, our study demonstrates that OH-CATH30, a naturally occurring antimicrobial peptide, selectively stimulates the innate immune response to protect against sepsis.
Inflammation and oxidative stress are closely related processes in the pathogenesis of various ocular diseases. Uveitis is a disorder of the uvea and ocular tissues that causes extreme pain, ...decreases visual acuity, and can eventually lead to blindness. The pharmacological functions of fucoxanthin, isolated from brown algae, induce a variety of therapeutic effects such as oxidative stress reduction and repression of inflammation reactions. However, the specific anti-inflammatory effects of fucoxanthin on pathogen-associated molecular pattern (PAMP) lipopolysaccharide-induced uveitis have yet to be extensively described. Therefore, the aim of present study was to investigate the anti-inflammatory effects of fucoxanthin on uveitis in rats. The results showed that fucoxanthin effectively enhanced the expression of nuclear factor erythroid 2-related factor 2 (Nrf2) in ocular tissues. Furthermore, fucoxanthin significantly increased the ocular activities of superoxide dismutase and decreased the levels of malondialdehyde stimulated by PAMP-induced uveitis. Ocular hypertension and the levels of inflammatory cells and proinflammatory cytokine tumor necrosis factor-alpha in the aqueous humor were alleviated with fucoxanthin treatment. Consequently, compared to the observed effects in lipopolysaccharide groups, fucoxanthin treatment significantly preserved iris sphincter innervation and pupillary function. Additionally, PAMP-induced corneal endothelial disruption was significantly inhibited by fucoxanthin treatment. Overall, these findings suggest that fucoxanthin may protect against inflammation from PAMP-induced uveitis by promoting the Nrf2 pathway and inhibiting oxidative stress.
Colorectal cancer (CRC) is one of the most common causes of death in Taiwan. Previous studies showed that
(AC) can treat poisoning, diarrhea, and various types of cancer. Therefore, we purified a ...novel ubiquinone derivative, AC009, and investigated its antitumor effects. Cell viability assays revealed that AC009 reduced the viability of several human CRC cell lines. AC009 treatment resulted in cell-cycle arrest/apoptosis, and these effects may occur via caspase and Bcl-2 signaling pathways. We demonstrated that AC009 could significantly inhibit in vivo tumor growth in xenograft mouse models. Using messenger RNA (mRNA) and microRNA (miRNA) microarrays, we found that
gene expression was also regulated by AC009, possibly through specific miRNAs. AC009 also reduced cancer stem-cell marker CD44
/CD24
expression and restored the tumor inhibition effect of cetuximab in
-mutant CRC. Moreover, we found that miRNA-27a could restore the tumor inhibition effect of cetuximab in
-mutant CRC cells. Taken together, our results suggest that AC009 has therapeutic potential against human wild-type and
-mutant CRC.
Abstract
Because most of animal viruses are enveloped, cytoplasmic entry of these viruses via fusion with cellular membrane initiates their invasion. However, the strategies in which host cells ...counteract cytoplasmic entry of such viruses are incompletely understood. Pore-forming toxin aerolysin-like proteins (ALPs) exist throughout the animal kingdom, but their functions are mostly unknown. In this study, we report that βγ-crystallin fused aerolysin-like protein and trefoil factor complex (βγ-CAT), an ALP and trefoil factor complex from the frog Bombina maxima, directly blocks enveloped virus invasion by interfering with cytoplasmic entry. βγ-CAT targeted acidic glycosphingolipids on the HSV type 1 (HSV-1) envelope to induce pore formation, as indicated by the oligomer formation of protein and potassium and calcium ion efflux. Meanwhile, βγ-CAT formed ring-like oligomers of ∼10 nm in diameter on the liposomes and induced dye release from liposomes that mimic viral envelope. Unexpectedly, transmission electron microscopy analysis showed that the βγ-CAT–treated HSV-1 was visibly as intact as the vehicle-treated HSV-1, indicating that βγ-CAT did not lyse the viral envelope. However, the cytoplasmic entry of the βγ-CAT–treated HSV-1 into HeLa cells was totally hindered. In vivo, topical application of βγ-CAT attenuated the HSV-1 corneal infection in mice. Collectively, these results uncovered that βγ-CAT possesses the capacity to counteract enveloped virus invasion with its featured antiviral-acting manner. Our findings will also largely help to illustrate the putative antiviral activity of animal ALPs.
This study is intended to assess the types of joints applied in the Joinery Category of the Regional Taiwan Skills Competitions (hereafter referred to as the Regional Competitions, including Northern ...Taiwan, Central Taiwan and Southern Taiwan). The authors use a self-developed derivation calculation formula (∑i=1nw×hi) to examine the differences in the types of joints applied. Then a regression equation is adopted to estimate the distribution of weight values of the joint types to calculate the assessed values of the joint types likely to be used in the Regional Competitions of the 51st Taiwan Skills Competition. Subsequently, the similarity of the fuzzy number is employed to understand the tendency in the development of joint types from the 36th Regional Competition to the 50th Regional Competition. The outcome shows that 19 types of joints were applied and the assessed value of 2.3 of open bevel mortise-tenon joints was the highest, followed by the 2 of bevel inclined mortise-tenon joints and the 0.73 of sloped haunch mortise-tenon joints. Meanwhile, the application of similarity of triangular fuzzy numbers to examine the 19 types of mortise-tenon joints indicates that the area of the affiliation map of open bevel mortise-tenon joints is the largest. Compared to the other 18 types of mortise-tenon joints, the assessed value of level of similarity is the lowest. In other words, the assessed values acquired by using the similarity of triangular fuzzy numbers and by using the derivation calculation formula have the same tendency. Hence, it is suggested that when trainers/instructors determine training strategies, they should adopt open bevel mortise-tenon joints, bevel inclined mortise-tenon joints and sloped haunch mortise-tenon joints to be the assessment standards. At the same time, they can also use the affiliation map of similarity of triangular fuzzy numbers to evaluate the differences between various types of mortise-tenon joints to understand the tendency of joint type development. Thus, the teaching quality of trainers/instructors and the learning results of contestants can both be upgraded. Systematic approaches also have to be consolidated to strengthen the ability to assess vocational education and training (VET) in order to improve the learning effects and technical capacity of Joinery category contestants. It is hoped that the derivation calculation formula can be applied to evaluate the training of Joinery category contestants in regional competitions to improve the quality of VET.
ABSTRACT
Tissue repair is a highly dynamic process, and the immediate onset of acute inflammation has been considered necessary for repair. Pore‐forming proteins are important, both in pathogen ...invasion and host immunity. However, their roles in wound healing and tissue repair are unclear. βγ‐crystallin fused aerolysin‐like protein (α‐subunit) and trefoil factor (β‐subunit) complex (βγ‐CAT) is a complex of a bacterial pore‐forming toxin aerolysin‐like protein and trefoil factor identified in the frog Bombina maxima. In this study, we established mouse cutaneous wound models to explore the effects of βγ‐CAT on skin wound healing. βγ‐CAT accelerated the healing of full‐thickness wounds by improving re‐epithelialization. This complex relieved dermal edema and promoted scarless healing. βγ‐CAT treatment resulted in a rapid release of IL‐1β, which initiated an acute inflammation response in the early stage of healing. Meanwhile, the expression levels of TGF‐β1, VEGF, and bFGF and the recruitment of M2 macrophages around the wound significantly increased after βγ‐CAT treatment. βγ‐CAT protected skin wounds against methicillin‐resistant Staphylococcus aureus by improving neutrophil recruitment at the site of the wound. Overall, our results suggest that βγ‐CAT can promote tissue repair and protect skin wounds against antibiotic‐resistant bacterial infection by triggering the acute inflammatory response. This is the first example that aerolysin‐like pore‐forming proteins widely existing in plants and animals may act in wound healing and tissue repair.—Gao, Z.‐H., Deng, C.‐J., Xie, Y.‐Y., Guo, X.‐L., Wang, Q.‐Q., Liu, L.‐Z., Lee, W.‐H., Li, S.‐A., Zhang, Y., Pore‐forming toxin‐like protein complex expressed by frog promotes tissue repair. FASEB J. 33, 782–795 (2019). www.fasebj.org
Our previous study showed that the extract of Inula japonica Thunb. (I. japonica) has anti-inflammatory and anti-asthmatic activities. In an attempt to find anti-inflammatory compounds from I. ...japonica, we recently isolated 1,6α-dihydroxy-4αH-1,10-secoeudesma-5(10),11(13)-dien-12,8β-olide (SE), 6α-isobutyryloxy-1-hydroxy-4αH-1,10-secoeudesma-5(10),11(13)-dien-12,8β-olide (IBSE), and 6α-isovaleryloxy-1-hydroxy-4αH-1,10-secoeudesma-5(10),11(13)-dien-12,8β-olide (IVSE) from the extract of I. japonica, and investigated their inhibitory effects on nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated RAW264.7 cells.
The inhibitory effect of IVSE, SE and IBSE on NO production in LPS-induced RAW264.7 cells was examined using Griess reagent, and the effects of IVSE on the expressions of inducible nitric oxide synthase (iNOS) and its upstream signal proteins including IκB kinase (IKK)/inhibitor kappa B (IκB)-α/nuclear factor κB (NF-κB) and mitogen-activated protein kinases (MAPKs) were investigated by Western blot.
Among the 3 compounds isolated, SE, IBSE, and IVSE inhibited NO production at 2.5μM with 5.1%, 40.4%, and 52.8%, respectively. IVSE displayed the most potent inhibition of NO production. Mechanism analysis indicated that IVSE dramatically decreased the expression of iNOS, reduced the translocation of the NF-κB subunit p65 into the nucleus by interrupting the phosphorylation and degradation of IκB-α, and inhibited the activation of the upstream mediator IKK α/β. Furthermore, our results showed that IVSE inhibited the phosphorylation of MAPKs including extracellular regulated kinases (ERK1/2), c-Jun N-terminal kinases (JNK) and p38.
IVSE exhibited anti-inflammatory activity by inhibiting NO production, in which inactivation of NF-κB and MAPKs might be involved. Our results suggest that IVSE might become an anti-inflammatory drug candidate.
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