Exploring the Relationship between Gut Microbiome Composition and Blood Indole-3-acetic Acid in Hemodialysis Patients.

Indole-3-acetic acid (IAA), a protein-bound uremic toxin resulting from gut microbiota-driven tryptophan metabolism, increases in hemodialysis (HD) patients. IAA may induce endothelial dysfunction, inflammation, and oxidative stress, elevating cardiovascular and cognitive risk in HD patients. However, research on the microbiome-IAA association is limited. This study aimed to explore the gut microbiome's relationship with plasma IAA levels in 72 chronic HD patients aged over 18 (August 2016-January 2017). IAA levels were measured using tandem mass spectrometry, and gut microbiome analysis utilized 16s rRNA next-generation sequencing. Linear discriminative analysis effect size and random forest analysis distinguished microbial species linked to IAA levels. Patients with higher IAA levels had reduced microbial diversity. Six microbial species significantly associated with IAA levels were identified; Bacteroides clarus, Bacteroides coprocola, Bacteroides massiliensi, and Alisteps shahii were enriched in low-IAA individuals, while Bacteroides thetaiotaomicron and Fusobacterium varium were enriched in high-IAA individuals. This study sheds light on specific gut microbiota species influencing IAA levels, enhancing our understanding of the intricate interactions between the gut microbiota and IAA metabolism.

Highly hydroxylated hafnium clusters are accessible to high resolution EUV photoresists under small energy doses.

This work reports the success in accessing high-resolution negative-tone EUV photoresists without radical chain growth in the aggregation mechanism. The synthesis of a highly hydroxylated Hf6O4(OH)8(RCO2)8 cluster 3 (R = s-butyl or s-Bu) is described; its EUV performance enables high resolution patterns HP = 18 nm under only 30 mJ cm-2. This photoresist also achieves high resolution patterns for e-beam lithography. Our new photoresist design to increase hydroxide substitutions of carboxylate ligands in the Hf6O4(OH)4(RCO2)12 clusters improves the EUV resolution and also greatly reduces EUV doses. Mechanistic analysis indicates that EUV light not only enables photolytic decomposition of carboxylate ligands, but also enhances the Hf-OH dehydration. One additional advantage of cluster 3 is a very small loss of film thickness (ca. 13%) after the EUV pattern development.

Unusual core engineering on a copper hydride nanoball.

A neutral polyhydrido copper cluster, [Cu27H15{S2CNnBu2}12] (abbreviated as [Cu27H15]), was prepared by the reaction of dithiocarbamates (dtc), Cu(I) salts and NaBH4. The isolated cluster provides insights into core engineering, demonstrating its novel ability to reversibly add or remove one copper atom from the cluster core. Single-crystal X-ray analysis reveals that the new core-shell structure exhibits a Cu24 rhombicuboctahedral outer cage and an inner Cu3 triangular kernel. The two core-shell clusters, [Cu27H15{S2CNnBu2}12] and previously published [Cu28H15(S2CNnBu2)12]+ (abbreviated as [Cu28H15]+), are only differentiated by one copper atom in their inner core. Importantly, we demonstrate core engineering with the controllable reversible transition between an irregular Cu4 tetrahedron and a Cu3 triangle, whilst maintaining their outer Cu24 shell intact. The 15 hydride atoms in [Cu27H15], coordinated in three different modes, are co-incident with the hydride positions in [Cu28H15]+. The degradation of [Cu27H15] in solution or the addition of one eq. of Cu(I) ions leads to the conversion of [Cu27H15] into [Cu28H15]+, while the reverse transformation can be achieved by the addition of either formic acid or a reducing agent to [Cu28H15]+. A dicationic species was observed in the ESI mass spectrum, and the composition is formulated as [Cu56H30(S2CNnBu2)24]2+, a dimer of [Cu27H15(S2CNnBu2)12 + Cu+]22+. The dimeric species was further explored by DFT calculations, suggesting that the lowest energy structure consists of a [Cu28H15]+ and a [Cu27H15] cluster connected through one Cu+ atom bridge. As a result, [Cu27H15] is considered an intermediate species in the formation of the more stable [Cu28H15]+ nanoball.

Verbascoside and isoverbascoside ameliorate transforming growth factor ß1-induced collagen expression by lung fibroblasts through Smad/non-Smad signaling pathways.

AIMS: Pulmonary fibrosis (PF) is a chronic, irreversible, and debilitating lung disease that typically leads to respiratory failure, and is a major cause of morbidity and mortality. Few drugs are effective for the treatment of patients with PF or for reducing the rate of disease progression. MAIN METHODS: Transforming growth factor-ß1 (TGF-ß1) is a profibrotic cytokine that signals through Smad and non-Smad pathways. Verbascoside (VB) and isoverbascoside (isoVB) exhibit anti-oxidative and anti-inflammatory activities, however, their anti-fibrotic effects remain unclear. This study evaluated the effects of VB and isoVB on TGF-ß1-stimulated murine lung fibroblasts (MLg 2908) and also human lung fibroblasts (confirmed by immunostaining). KEY FINDINGS: Neither VB nor isoVB had a cytotoxic effect on MLg 2908 fibroblasts. Both compounds (10 µM) reduced intracellular reactive oxygen species and markedly attenuated collagen I expression in TGF-ß1 (5 ng/ml)-induced MLg 2908 cells compared to TGF-ß1 alone. Both compounds suppressed the TGF-ß1-induced phosphorylation of Smad2/3 and ERK/p38 mitogen-activated protein kinases (MAPKs). VB and isoVB, but not pirfenidone and nintedanib, inhibited TGF-ß1-induced pSmad2/3, ERK/p38 MAPK, and collagen I expression. VB and isoVB also decreased collagen I deposition in TGF-ß1-induced MLg 2908 cells. Only isoVB significantly suppressed collagen I deposition in TGF-ß1-induced human pulmonary cells. Our results indicated that VB and isoVB may exert antifibrotic effects by inhibiting TGF-ß1-induced collagen I expression via inhibition of oxidative stress and downregulation of the Smad/non-Smad pathway. SIGNIFICANCE: The present findings suggest that VB or isoVB may be used as a supplement to alleviate PF.

Coenzyme Q0 Inhibits NLRP3 Inflammasome Activation through Mitophagy Induction in LPS/ATP-Stimulated Macrophages.

Coenzyme Q (CoQ) analogs with a variable number of isoprenoid units have exhibited as anti-inflammatory as well as antioxidant molecules. Using novel quinone derivative CoQ0 (2,3-dimethoxy-5-methyl-1,4-benzoquinone, zero side chain isoprenoid), we studied its molecular activities against LPS/ATP-induced inflammation and redox imbalance in murine RAW264.7 macrophages. CoQ0's non- or subcytotoxic concentration suppressed the NLRP3 inflammasome and procaspase-1 activation, followed by downregulation of IL1ß expression in LPS/ATP-stimulated RAW264.7 macrophages. Similarly, treatment of CoQ0 led to LC3-I/II accumulation and p62/SQSTM1 activation. An increase in the Beclin-1/Bcl-2 ratio and a decrease in the expression of phosphorylated PI3K/AKT, p70 S6 kinase, and mTOR showed that autophagy was activated. Besides, CoQ0 increased Parkin protein to recruit damaged mitochondria and induced mitophagy in LPS/ATP-stimulated RAW264.7 macrophages. CoQ0 inhibited LPS/ATP-stimulated ROS generation in RAW264.7 macrophages. Notably, when LPS/ATP-stimulated RAW264.7 macrophages were treated with CoQ0, Mito-TEMPO (a mitochondrial ROS inhibitor), or N-acetylcysteine (NAC, a ROS inhibitor), there was a significant reduction of LPS/ATP-stimulated NLRP3 inflammasome activation and IL1ß expression. Interestingly, treatment with CoQ0 or Mito-TEMPO, but not NAC, significantly increased LPS/ATP-induced LC3-II accumulation indicating that mitophagy plays a key role in the regulation of CoQ0-inhibited NLRP3 inflammasome activation. Nrf2 knockdown significantly decreased IL1ß expression in LPS/ATP-stimulated RAW264.7 macrophages suggesting that CoQ0 inhibited ROS-mediated NLRP3 inflammasome activation and IL1ß expression was suppressed due to the Nrf2 activation. Hence, this study showed that CoQ0 might be a promising candidate for the therapeutics of inflammatory disorders due to its effective anti-inflammatory as well as antioxidant properties.

Star-ECG: Visualization of Electrocardiograms for Arrhythmia and Heart Rate Variability.

Conventional electrocardiograms (ECG) are displayed in one dimension. Reading one-dimensional ECG waveform becomes challenging when one wants to visualize the heart rate variability with naked eye. Some ECG visualization techniques have been proposed. However, they rely on domain knowledge to comprehend the heart rate variability. To improve the readability for patients and non-experts, we introduce Star-ECG, a novel ECG visualization approach. Such approach projects ECG waveforms onto a two-dimensional plane in a circular form. We demonstrate that Star-ECG offers not only easily deciphered visualization of cardiac abnormalities and heart rate variability, but also the application of state-of-the-art arrhythmia classification with integrated deep neural networks. We also report positive user feedback from both experts and non-experts that Star-ECG can provide readable and helpful information to monitor cardiac activities.

Sweet Olive Extract Attenuates Dextran Sulfate Sodium-Induced Colitis in Mice.

Sweet olive (Osmanthus fragrans flowers) is used to treat dysentery and reduce phlegm and stasis in traditional Chinese medicine. Recently, we found that verbascoside, the major component in the sweet olive ethanolic extract (OFE), inhibited IL-8 secretion in human colorectal adenocarcinoma WiDr cells. However, evidence-based treatment of inflammatory bowel disease (IBD) with the extract is yet to be performed. To evaluate the therapeutic effect of OFE, we measured IL-8 suppression by OFE and verbascoside in a WiDr cell culture assay. In the IL-8 secretion assay, both OFE (100 µg/mL) and verbascoside (10 µM) significantly inhibited IL-8 production in WiDr cells. Furthermore, we designed cotreated (dextran sulfate sodium [DSS]+OFE-treated) and post-treated (DSS-OFE-treated) protocols to access the therapeutic effects of OFE in vivo. Mice treated with 500 mg/kg per day OFE exhibited significant improvement in IBD symptoms, including disease activity index score, body weight, and colon length maintenance. The suppressive effects on myeloperoxidase expression and lower histopathology scores (including neutrophil infiltration) for the colon were also found. These findings suggest that OFE exerts anti-inflammatory effect on DSS-induced colitis.

Using Flow Cytometry to Isolate Maize Meiocytes for Next Generation Sequencing: A Time and Labor Efficient Method.

Meiosis is essential during sexual reproduction to generate haploid gametes. Genomic or epigenomic studies of meiosis in multicellular organisms using next-generation sequencing (NGS) methods have been limited because of the difficulty of collecting thousands to millions of meiocytes. Here, we describe a simple protocol to efficiently isolate maize male meiocytes from formaldehyde-fixed samples for NGS techniques that require chemical crosslinking to preserve complex interactions or chromatin architecture. Anthers at desired meiotic stages are selected, fixed with paraformaldehyde, and disrupted using a homogenizer. Cell walls are digested to produce a cell suspension containing small somatic cells and large individual meiocytes. The meiocyte fraction is enriched by size separation with cell strainers and further purified by flow cytometry. From 400 anthers, we can isolate 20,000 meiocytes at 98% purity in 6 to 8 hours. © 2018 by John Wiley & Sons, Inc.

Predictors of smoking cessation in Taiwan: using the theory of planned behavior.

This study aimed to explore the factors predicting the intention to quit smoking and the subsequent behavior 6 months later using the theory of planned behavior (TPB). Data were obtained from 145 smokers who attended a smoking cessation clinic in a community hospital. All participants completed a questionnaire which included demographic information, TPB-based items, perceived susceptibility and previous attempts to quit. The actual quitting behavior was obtained by follow-up phone calls 6 months later. The TPB constructs explained 34% of the variance in intention to quit smoking. By adding perceived susceptibility, the explained variance was significantly improved to 40%. The most important predictors were perceived behavior control and perceived susceptibility, followed by attitude. Subjective norm did not contribute to the prediction of intention. Attitude and perceived behavior control contributed to the prediction of actual quitting behavior, but intention, subjective norm and perceived susceptibility did not. Our findings support that the TPB is generally a useful framework to predict the intention to quit smoking in Taiwan. The inclusion of perceived susceptibility improved the prediction of intention. With regards to successfully quitting, attitude and perceived behavior control played more crucial roles than other TPB constructs. Smoking cessation promotion initiatives focusing on reinforcing cessation belief, enhancing a smoker's perception of their capability to quit smoking, and persuading smokers that they can overcome cessation barriers to cessation could make subsequent interventions more effective.

Geographical variations in bacterial communities associated with soft coral Scleronephthya gracillimum.

Environmental impacts can alter relationships between a coral and its symbiotic microbial community. Furthermore, changes in the microbial community associated with increased seawater temperatures can cause opportunistic infections, coral disease and death. Interactions between soft corals and their associated microbes are not well understood. The species Scleronephthya gracillimum is distributed in tropical to temperate zones in coral assemblages along the Kuroshio Current region. In this study we collected S. gracillimum from various sites at different latitudes, and compared composition of their bacterial communities using Next Generation Sequencing. Coral samples from six geographically distinct areas (two sites each in Taiwan, Japan, and Korea) had considerable variation in their associated bacterial communities and diversity. Endozoicimonaceae was the dominant group in corals from Korea and Japan, whereas Mycoplasma was dominant in corals from Taiwan corals. Interestingly, the latter corals had lower relative abundance of Endozoicimonaceae, but greater diversity. These biogeographic differences in bacterial composition may have been due to varying environmental conditions among study locations, or because of host responses to prevailing environmental conditions. This study provided a baseline for future studies of soft coral microbiomes, and assessment of functions of host metabolites and soft coral holobionts.

Suppression of IL-8 Release by Sweet Olive Ethanolic Extract and Compounds in WiDr Colon Adenocarcinoma Cells.

Oxidative stress can stimulate the secretion of pro-inflammatory cytokines. Interleukin-8 (IL-8) has been implicated in the pathogenesis of inflammatory bowel disease and the metastatic spread of colorectal cancer. The flowers of Osmanthus fragrans (sweet olive) are used to alleviate dysentery with blood in the bowel, as well as stomach ache and diarrhea. However, the evidence of their therapeutic effects on these symptoms remains unclear. In the present study, the protective effects of sweet olive flower ethanolic extract (OFE) against oxidative stress in WiDr cells was assessed by evaluating its 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity. In addition, cellular IL-8 secretion was evaluated. Notably, high-performance liquid chromatography showed verbascoside to be the primary constituent in OFE; it exhibited a DPPH scavenging activity with an IC50 of 8.23 µg/mL. Moreover, OFE (1 to 100 µg/mL) showed a potent, dose-dependent inhibitory effect on H2 O2 -induced IL-8 secretion in WiDr cells. Nine compounds were isolated from OFE based on a protective effect-guided purification process. Of these compounds, 5 phenolic compounds-verbascoside, phillygenin, tyrosol, methyl 4-hydroxycinnamate, and eutigoside A-reduced IL-8 secretion at 10 µg/mL treatment concentrations. Further analysis showed that the anti-inflammatory effects of OFE likely occurred via nuclear factor-κB pathway inhibition, which attenuates IL-8 secretion in cells. Collectively, these data suggest that OFE could be developed as an agent that suppresses IL-8 secretion to treat chronic inflammatory diseases.

Frequency and pattern of Chinese herbal medicine prescriptions for urticaria in Taiwan during 2009: analysis of the national health insurance database.

BACKGROUND: Large-scale pharmaco-epidemiological studies of Chinese herbal medicine (CHM) for treatment of urticaria are few, even though clinical trials showed some CHM are effective. The purpose of this study was to explore the frequencies and patterns of CHM prescriptions for urticaria by analysing the population-based CHM database in Taiwan. METHODS: This study was linked to and processed through the complete traditional CHM database of the National Health Insurance Research Database in Taiwan during 2009. We calculated the frequencies and patterns of CHM prescriptions used for treatment of urticaria, of which the diagnosis was defined as the single ICD-9 Code of 708. Frequent itemset mining, as applied to data mining, was used to analyse co-prescription of CHM for patients with urticaria. RESULTS: There were 37,386 subjects who visited traditional Chinese Medicine clinics for urticaria in Taiwan during 2009 and received a total of 95,765 CHM prescriptions. Subjects between 18 and 35 years of age comprised the largest number of those treated (32.76%). In addition, women used CHM for urticaria more frequently than men (female:male = 1.94:1). There was an average of 5.54 items prescribed in the form of either individual Chinese herbs or a formula in a single CHM prescription for urticaria. Bai-Xian-Pi (Dictamnus dasycarpus Turcz) was the most commonly prescribed single Chinese herb while Xiao-Feng San was the most commonly prescribed Chinese herbal formula. The most commonly prescribed CHM drug combination was Xiao-Feng San plus Bai-Xian-Pi while the most commonly prescribed triple drug combination was Xiao-Feng San, Bai-Xian-Pi, and Di-Fu Zi (Kochia scoparia). CONCLUSIONS: In view of the popularity of CHM such as Xiao-Feng San prescribed for the wind-heat pattern of urticaria in this study, a large-scale, randomized clinical trial is warranted to research their efficacy and safety.

Significant relationship between serum high-sensitivity C-reactive protein, high-density lipoprotein cholesterol levels and children with Kawasaki disease and coronary artery lesions.

BACKGROUND/PURPOSE: The aim of this study was to understand the association between Kawasaki disease (KD) in children and serum high-sensitivity C-reactive protein (hs-CRP), which is a sensitive indicator of inflammation, lipid profiles and coronary artery lesions. METHODS: Between July 2005 and December 2007, 119 children with KD at least 1 year after diagnosis were recruited. The children were classified into one of two groups: Group I comprised 55 children with KD and coronary aneurysms; Group 2 comprised 64 children with KD and normal coronary arteries. The relationship between hs-CRP concentration and high-density lipoprotein cholesterol (HDL-C) and the size of arterial lesions was investigated in Group I 1 year after onset of KD. RESULTS: Serum hs-CRP levels in Group I (mean, 0.251 mg/dL) were significantly greater than those in Group II (mean, 0.162 mg/dL; p = 0.011). However, plasma HDL-C levels in Group I (mean 42.51 mg/dL) were significantly lower than those in Group II (mean, 44.34 mg/dL; p = 0.037). In Group I, there was a positive association between hs-CRP and the size of coronary artery lesions (r = 0.672, p = 0.035), but no association between lipid profiles, including HDL-C, and coronary artery lesions (all p > 0.05). CONCLUSION: Our results support the possibility of ongoing low-grade inflammation late after the acute phase of KD in children with coronary aneurysms. Serum hs-CRP and HDL-C levels are associated with coronary artery lesions in children with KD.

The role of Mycoplasma pneumoniae in acute exacerbation of asthma in children.

BACKGROUND: The aim of this retrospective study was to test the connection between acute M. pneumoniae infection and the exacerbation of asthma. The clinical characteristics of Mycoplasma infection seen during emergent visits in asthmatic children were reviewed. METHODS: We examined the cases of 316 asthma exacerbation patients aged from two to fourteen-years-old. They were divided into two groups according to their asthma history. One hundred and eighty-eight cases had evidence of a history of asthma (group 1) and 128 had only had their first asthma attack (group 2). The control group (group 3) was made up of 151 asthmatic children who had no acute exacerbation during the previous six months. In all three groups, we looked whether those children had acute M. pneumoniae infection or not. Acute M. pneumoniae infection was defined by positive results in serologic testing, with specific immunoglobulin M (IgM) antibody or with a > or = fourfold increase in IgG titer by the third week in the same children. RESULTS: In group 1, acute M. pneumionae infection was found in 42 (23%) of the 188 children. In group 2, acute M. pneumoniae infection was proven in 57 (45%) of the 128 children. In the control group, 10 (7%) of the 151 children had M. pneumoniae infection. Twenty-three (54%) asthmatic children that presented with fever as the chief complaint were infected with M. pneumoniae, compared with 18 (12%) children without infection (p = 0.014). Twenty-nine (50%) children having their first asthma attack with fever were infected with M. pneumoniae, compared with 22 (32%) without infection (p = 0.009). In group 1, 17 (41%) children with M. pneumoniae infections and 28 (19%) children without M. pneumoniae infections presented with rale breathing sounds of the physical examination (p = 0.027). In group 2, 26 (46%) children with M. pneumoniae infections and 17 (24%) children without M. pneumoniae infections presented with rale breathing sounds (p = 0.019). CONCLUSIONS: We found that M. pneumoniae may play a role in asthmatic exacerbation among children, especially in those experiencing their first asthma attack. Moreover, among children with acute M. pneumoniae infection, the number was significantly increased of children having fever as the chief complaint and rales in auscultations compared with those without M. pneumoniae infection.

Retaining of the assembly capability of vimentin phosphorylated by mitogen-activated protein kinase-activated protein kinase-2.

Intermediate filament (IF) networks can be regulated by phosphorylation of unit proteins, such as vimentin, by specific kinases leading to reorganization of the IF filamentous structure. Recently, we identified mitogen-activated protein kinase-activated protein kinase-2 (MAPKAP kinase-2) as a vimentin kinase (Cheng and Lai [1998] J. Cell. Biochem. 71:169-181). Herein we describe the results of further in vitro studies investigating the effects of MAPKAP kinase-2 phosphorylation on vimentin and the effects of the phosphorylation on the filamentous structure. We show that MAPKAP kinase-2 mainly phosphorylates vimentin at Ser-38, Ser-50, Ser-55, and Ser-82, residues all located in the head domain of the protein. Surprisingly, and in stark contrast to phosphorylation by most other kinases, phosphorylation of vimentin by MAPKAP kinase-2 has no discernable effect on its assembly. It suggested that structure disassembly is not the only obligated consequence of phosphorylated vimentin as regulated by other kinases. Finally, a mutational analysis of each of the phosphorylated serine residues in vimentin suggested that no single serine site was primarily responsible for structure maintenance, implying that the retention of filamentous structure may be the result of the coordinated action of several phosphorylated serine sites. This also shed new lights on the functional task(s) of vimentin that is intermediate filament proteins might provide a phosphate reservoir to accommodate the phosphate surge without any structural changes.