Chemical Probes and Activity-Based Protein Profiling for Cancer Research.

Chemical probes can be used to understand the complex biological nature of diseases. Due to the diversity of cancer types and dynamic regulatory pathways involved in the disease, there is a need to identify signaling pathways and associated proteins or enzymes that are traceable or detectable in tests for cancer diagnosis and treatment. Currently, fluorogenic chemical probes are widely used to detect cancer-associated proteins and their binding partners. These probes are also applicable in photodynamic therapy to determine drug efficacy and monitor regulating factors. In this review, we discuss the synthesis of chemical probes for different cancer types from 2016 to the present time and their application in monitoring the activity of transferases, hydrolases, deacetylases, oxidoreductases, and immune cells. Moreover, we elaborate on their potential roles in photodynamic therapy.

RNA-sequencing identification and validation of genes differentially expressed in high-risk adenoma, advanced colorectal cancer, and normal controls.

Distinct gene expression patterns that occur during the adenoma-carcinoma sequence need to be determined to analyze the underlying mechanism in each step of colorectal cancer progression. Elucidation of biomarkers for colorectal polyps that harbor malignancy potential is important for prevention of colorectal cancer. Here, we use RNA sequencing to determine gene expression profile in patients with high-risk adenoma treated with endoscopic submucosal dissection by comparing with gene expression in patients with advanced colorectal cancer and normal controls. We collected 70 samples, which consisted of 27 colorectal polyps, 24 cancer tissues, and 19 normal colorectal mucosa. RNA sequencing was performed on an Illumina platform to select differentially expressed genes (DEGs) between colorectal polyps and cancer, polyps and controls, and cancer and normal controls. The Kyoto Gene and Genome Encyclopedia (KEGG) and gene ontology (GO) analysis, gene-concept network, GSEA, and a decision tree were used to evaluate the DEGs. We selected the most highly expressed genes in high-risk polyps and validated their expression using real-time PCR and immunohistochemistry. Compared to patients with colorectal cancer, 82 upregulated and 24 downregulated genes were detected in high-risk adenoma. In comparison with normal controls, 33 upregulated and 79 downregulated genes were found in high-risk adenoma. In total, six genes were retrieved as the highest and second highest expressed in advanced polyps and cancers among the three groups. Among the six genes, ANAX3 and CD44 expression in real-time PCR for validation was in good accordance with RNA sequencing. We identified differential expression of mRNAs among high-risk adenoma, advanced colorectal cancer, and normal controls, including that of CD44 and ANXA3, suggesting that this cluster of genes as a marker of high-risk colorectal adenoma.

Intestinal Epithelial Deletion of Sphk1 Prevents Colitis-Associated Cancer Development by Inhibition of Epithelial STAT3 Activation.

BACKGROUND AND AIMS: Colitis-associated cancer (CAC) is one of the most serious complications in patients with inflammatory bowel disease. Sphingosine kinase 1 (Sphk1) is a key enzyme in the sphingolipid pathway and has oncogene potential for inducing both initiation and progression of tumors. The aim of this work is to characterize the role of epithelial Sphk1 in mouse colitis and CAC models. METHODS: We investigated the roles of Sphk1 in CAC by conditional deletion of Sphk1 in intestinal epithelial cells (IECs). RESULTS: CAC was induced in both Sphk1ΔIEC/ApcMin/+ and Sphk1IEC/ApcMin/+ mice by administration of 2% dextran sodium sulfate (DSS) for 7 days. Genetic deletion of Sphk1 significantly reduced the number and size of tumors in ApcMin/+ mice. Histologic grade was more severe in Sphk1ΔIEC/ApcMin/+ mice compared with Sphk1IEC/ApcMin/+ mice (invasive carcinoma, 71% versus 13%, p < 0.05). Deletion of Sphk1 decreased mucosal proliferation and inhibited STAT3 activation and genetic expression of cyclin D1 and cMyc in tumor cells. Conditional deletion of Sphk1 using CRISPR-Cas9 in HCT 116 cells inhibited interleukin (IL)-6-mediated STAT3 activation. CONCLUSIONS: Epithelial conditional deletion of Sphk1 inhibits CAC in ApcMin/+-DSS models in mice by inhibiting STAT3 activation and its target signaling pathways.

Recycling rice husks for high-capacity lithium battery anodes.

The rice husk is the outer covering of a rice kernel and protects the inner ingredients from external attack by insects and bacteria. To perform this function while ventilating air and moisture, rice plants have developed unique nanoporous silica layers in their husks through years of natural evolution. Despite the massive amount of annual production near 10(8) tons worldwide, so far rice husks have been recycled only for low-value agricultural items. In an effort to recycle rice husks for high-value applications, we convert the silica to silicon and use it for high-capacity lithium battery anodes. Taking advantage of the interconnected nanoporous structure naturally existing in rice husks, the converted silicon exhibits excellent electrochemical performance as a lithium battery anode, suggesting that rice husks can be a massive resource for use in high-capacity lithium battery negative electrodes.

Design and fabrication of new nanostructured SnO2-carbon composite microspheres for fast and stable lithium storage performance.

One-pot method for metal oxide-carbon composite microsphere with three-dimensional ordered macroporous (3DOM) structure is first introduced. The 3DOM structured SnO2 -carbon microspheres prepared as the first target material show superior electrochemical properties as anode material for lithium ion batteries. The newly developed process can be applied to the preparation of 3DOM-structured metal oxide-carbon composite microspheres for wide applications.

Inhibitory effect of putranjivain A on allergic inflammation through suppression of mast cell activation.

A great number of people are suffering from allergic inflammatory disease such as asthma, atopic dermatitis, and sinusitis. Therefore discovery of drugs for the treatment of these diseases is an important subject in human health. Putranjivain A (PJA), member of ellagitannin, is known to possess beneficial effects including anti-cancer and anti-viral activities. The aim of the present study was to elucidate whether PJA modulates the allergic inflammatory reaction and to study its possible mechanisms of action using mast cell-based in vitro and in vivo models. The study was performed in anaphylaxis mouse model and cultured mast cells. PJA inhibited the expression of pro-inflammatory cytokines in immunoglobulin E-stimulated mast cells. PJA reduced this expression by inhibiting nuclear factor (NF)-κB and nuclear factor of activated T cell. The oral administration of PJA reduced systemic and cutaneous anaphylaxis, the release of serum histamine, and the expression of the histamine H1 receptor. In addition, PJA attenuated the activation of mast cells. PJA inhibited the release of histamine from various types of mast cells by the suppression of intracellular calcium. The inhibitory activity of PJA on the allergic reaction was similar to that of disodium cromoglycate, a known anti-allergic drug. These results suggest that PJA can facilitate the prevention or treatment of allergic inflammatory diseases mediated by mast cells.

Spray drying method for large-scale and high-performance silicon negative electrodes in Li-ion batteries.

Nanostructured silicon electrodes have shown great potential as lithium ion battery anodes because they can address capacity fading mechanisms originating from large volume changes of silicon alloys while delivering extraordinarily large gravimetric capacities. Nonetheless, synthesis of well-defined silicon nanostructures in an industrially adaptable scale still remains as a challenge. Herein, we adopt an industrially established spray drying process to enable scalable synthesis of silicon-carbon composite particles in which silicon nanoparticles are embedded in porous carbon particles. The void space existing in the porous carbon accommodates the volume expansion of silicon and thus addresses the chronic fading mechanisms of silicon anodes. The composite electrodes exhibit excellent electrochemical performance, such as 1956 mAh/g at 0.05C rate and 91% capacity retention after 150 cycles. Moreover, the spray drying method requires only 2 s for the formation of each particle and allows a production capability of ~10 g/h even with an ultrasonic-based lab-scale equipment. This investigation suggests that established industrial processes could be adaptable to the production of battery active materials that require sophisticated nanostructures as well as large quantity syntheses.

One-pot facile synthesis of ant-cave-structured metal oxide-carbon microballs by continuous process for use as anode materials in Li-ion batteries.

This paper introduces a facile one-pot method for synthesizing a new structured material, named "ant-cave microball", by continuous ultrasonic spray pyrolysis. The ant-cave-structured microballs are prepared from a colloidal spray solution with polystyrene nanobeads and sucrose. Networking between the nanovoids formed by decomposition of the polystyrene nanobeads results in the formation of nanochannels. The electrochemical properties of these ant-cave-structured MoO3-C microballs, prepared as the first target material for lithium ion batteries, are investigated. The nanochannels are uniformly distributed inside the microballs with MoO3 and carbon components uniformly distributed within the microballs. Further, the microballs have initial discharge and charge capacities of 1212 and 841 mA h g(-1), respectively, at a current density of 2 A g(-1), and the initial discharge and charge capacities based on the weight of MoO3 (disregarding carbon component) are as high as 1814 and 1259 mA h g(-1). The microballs deliver a high discharge capacity of 733 mA h g(-1) even after 300 cycles. This is although microsized MoO3 powders with a filled structure have discharge capacities of 1256 and 345 mA h g(-1) for the first and 300th cycles, respectively.

p,p'-DDT induces apoptosis in human endometrial stromal cells via the PI3K/AKT pathway and oxidative stress.

Objective: Bis-[4-chlorophenyl]-1,1,1-trichloroethane (DDT), one of the most widely used synthetic pesticides, is an endocrine-disrupting chemical with the potential to interfere with the human reproductive system. The effects of DDT and one of its metabolites, p,p'-DDT, on human endometrial stromal cells (ESCs) and health outcomes remain unknown. In this study, we investigated whether p,p'-DDT induces an imbalance in cell proliferation and apoptosis in human ESCs via oxidative stress. Methods: We assessed apoptosis in ESCs by quantifying the expression of markers associated with both intrinsic and extrinsic pathways. Additionally, we measured levels of reactive oxygen species (ROS), antioxidant enzyme activity, and estrogen receptors (ERs). We also examined changes in signaling involving nuclear factor kappa-light-chain-enhancer of activated B cells. Results: Following treatment with 1,000 pg/mL of p,p'-DDT, we observed an increase in Bax expression, a decrease in Bcl-2 expression, and increases in the expression of caspases 3, 6, and 8. We also noted a rise in the generation of ROS and a reduction in glutathione peroxidase expression after treatment with p,p'-DDT. Additionally, p,p'-DDT treatment led to changes in ER expression and increases in the protein levels of phosphatidylinositol 3-kinase (PI3K), phospho-protein kinase B (phospho-AKT), and phospho-extracellular signal-regulated kinase (phospho-ERK). Conclusion: p,p'-DDT was found to induce apoptosis in human ESCs through oxidative stress and an ER-mediated pathway. The activation of the PI3K/AKT and ERK pathways could represent potential mechanisms by which p,p'-DDT prompts apoptosis in human ESCs and may be linked to endometrial pathologies.

Solution processed aluminum paper for flexible electronics.

As an alternative to vacuum deposition, preparation of highly conductive papers with aluminum (Al) features is successfully achieved by the solution process consisting of Al precursor ink (AlH(3){O(C(4)H(9))(2)}) and low temperature stamping process performed at 110 °C without any serious hydroxylation and oxidation problems. Al features formed on several kinds of paper substrates (calendar, magazine, and inkjet printing paper substrates) are less than ~60 nm thick, and their electrical conductivities were found to be as good as thermally evaporated Al film or even better (≤2 Ω/□). Strong adhesion of Al features to paper substrates and their excellent flexibility are also experimentally confirmed by TEM observation and mechanical tests, such as tape and bending tests. The solution processed Al features on paper substrates show different electrical and mechanical performance depending on the paper type, and inkjet printing paper is found to be the best substrate with high and stable electrical and mechanical properties. The Al conductive papers produced by the solution process may be applicable in disposal paper electronics.

Triphenyl phosphate activates estrogen receptor α/NF-κB/ cyclin D1 signaling to stimulate cell cycle progression in human Ishikawa endometrial cancer cells.

OBJECTIVE: Triphenyl phosphate (TPHP) is one of the most commonly used organophosphorus flame retardants that may accumulate in the environment. However, its effects on human reproductive organs have not been well studied. We aimed to investigate the in vitro effects of TPHP in human Ishikawa endometrial cancer cells to elucidate how TPHP exposure disrupts intracellular signaling and cell proliferation in reproductive tissues. METHODS: Human Ishikawa endometrial cancer cells were exposed to TPHP. RESULTS: Exposure to TPHP elevated the levels of estrogen receptor (ER) α and progesterone receptor-B and reduced ER ß in human Ishikawa endometrial cancer cells. TPHP stimulated phosphoinositide 3-kinase/protein kinase B and mitogenactivated protein kinase/ extracellular signal-regulated kinases 1/2 kinase signaling, which may contribute to the activation of ER function and induce nuclear translocation of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) in human Ishikawa endometrial cancer cells. Activated ER and NF-κB stimulate the expression of cyclin D1/ cyclin-dependent kinase (CDK) 4/CDK6, indicating cell cycle progression and proliferation. CONCLUSION: This report may provide new information on the molecular mechanisms underlying how TPHP exposure dysregulates the cellular physiology of the human endometrium.

Dabigatran Acylglucuronide, the Major Metabolite of Dabigatran, Shows a Weaker Anticoagulant Effect than Dabigatran.

Dabigatran (DAB) is an orally administered thrombin inhibitor. Both DAB and its main metabolite dabigatran acylglucuronide (DABG) have established anticoagulant effects. Here, we aimed to compare the relative anticoagulant effects of DABG and DAB in humans. Anticoagulant effects of DAB and DABG were measured in vitro using a thrombin generation assay. Additionally, their effects on other coagulation assays including PT, aPTT, TT, and fibrinogen were compared. Both DAB and DABG showed inhibitory effects on thrombin generation in a dose-dependent manner, but DABG exhibited a weaker inhibitory effect than that of DAB. The IC50 values of DAB and DABG on thrombin generation AUC were 134.1 ng/mL and 281.9 ng/mL, respectively. DABG also exhibited weaker anticoagulant effects than DAB on PT, aPTT, and TT. The results of the present study indicate that the anticoagulant effect of DABG, a main active DAB metabolite, is weaker than that of DAB.

TMI-1, TNF-α-Converting Enzyme Inhibitor, Protects Against Paclitaxel-Induced Neurotoxicity in the DRG Neuronal Cells In Vitro.

Background: Chemotherapy-induced peripheral neuropathy (CIPN) negatively impacts cancer survivors' quality of life and is challenging to treat with existing drugs for neuropathic pain. TNF-α is known to potentiate TRPV1 activity, which contributes to CIPN. Here, we assessed the role of TMI-1, a TNF-α-converting enzyme inhibitor, in paclitaxel (PAC)-induced neurotoxicity in dorsal root ganglion (DRG) cells. Materials and Methods: Immortalized DRG neuronal 50B11 cells were cultured and treated with PAC or PAC with TMI-1 following neuronal differentiation. Cell viability, analysis of neurite growth, immunofluorescence, calcium flow cytometry, western blotting, quantitative RT-PCR, and cytokine quantitation by ELISA were performed to determine the role of TMI-1 in neurotoxicity in neuronal cells. Results: PAC administration decreased the length of neurites and upregulated the expression of TRPV1 in 50B11 cells. TMI-1 administration showed a protective effect by suppressing inflammatory signaling, and secretion of TNF-α. Conclusion: TMI-1 partially protects against paclitaxel-induced neurotoxicity by reversing the upregulation of TRPV1 and decreasing levels of inflammatory cytokines, including TNF-α, IL-1ß, and IL-6 in neuronal cells.

Synthesis of high thermally-stable mesoporous alumina particles.

The mesoporous undoped and Si-doped alumina were prepared with an ultrasonic spray process, and found to have well-developed mesopore structures and large surface areas. The mesoporous Si-doped alumina has a high thermal stability up to 1473 K. Its surface area and pore volume were found to slowly decrease with increasing temperature. Mesoporous undoped alumina is transformed to gamma-alumina at 1073 K, whereas the amorphous nature of the pore walls of the Si-doped alumina is maintained up to 1073 K. When heat treatment was carried out at 1473 K for 2 h, the mesopore-networks of the undoped alumina collapsed, and then all the pore walls were converted into the alpha-alumina phase. In contrast, the mesoporosity of the Si-doped alumina persisted during heat treatment, and its pore walls were transformed to gamma-alumina. The decreases in the pore volume of the undoped alumina at 1073 K and 1473 K were found to be 36% and 99% respectively, but for the Si-doped alumina were only 24% and 36% respectively. The surface area of the undoped alumina at 1473 K was found to be 11 m2/g but that of the Si-doped samples at the same temperature is higher than 100 m2/g. Thus this mesoporous Si-doped alumina can be used as a catalytic support in reactions at high temperatures.

1,2,3,6-tetra-O-galloyl-beta-D-allopyranose gallotannin isolated, from Euphorbia jolkini, attenuates LPS-induced nitric oxide production in macrophages.

Nitric oxide (NO) is a pleiotropic regulator, critical to numerous biological processes, including vasodilatation and macrophage-mediated immunity. Macrophages express inducible NO synthase (iNOS) and produce NO after lipopolysaccharide (LPS) stimulation. Gallotannins are water-soluble polyphenols with wide-ranging biological activities. Various chemical structures of gallotannins occurring in medicinal and food plants that are used worldwide showed several remarkable biological and pharmacological activities. In the present study, we examined the inhibitory effects of gallotannin 1,2,3,6-tetra-O-galloyl-beta-D-allopyranose (GT24) isolated from Euphorbia jolkini on the LPS-induced NO production and underlying mechanisms of action. GT24 dose-dependently decreased LPS-induced NO production and iNOS expression in J774A.1 macrophages. In addition, GT24 inhibited LPS-induced activation of nuclear factor (NF)-kappaB as indicated by inhibition of degradation of I-kappaBalpha, nuclear translocation of NF-kappaB, and NF-kappaB dependent gene reporter assay. Our results suggest that GT24 possesses an inhibitory effect on the LPS-induced inflammatory reaction.

Toxic effects of mercuric sulfide on immune organs in mice.

Mercuric sulfide (HgS) is a major component of cinnabar, which has been used as a sedative drug in China for more than 2000 years. Because its toxicological effects are still unclear, we attempted to verify the toxic effects of HgS, focused on liver and immune organs such as the spleen and thymus. Male ICR mice were administered HgS (0.02, 0.2, 2.0 g/kg/day) by gavage for 4 weeks. During the administration period, HgS-treated mice did not reveal overt signs of clinical toxicity. HgS had no significant effect on body weight, food consumption, water consumption, and organ weights. In spite of its known insolubility, HgS was absorbed by the gastrointestinal tract and accumulated in the liver, spleen and thymus in a dose-dependent manner. In the biochemical and histological examination, HgS did not cause hepatotoxicity. However, HgS significantly increased both CD8(+) T lymphocytes and CD4(+)CD8(+) lymphocyte populations in the spleen without changing in the thymus. In the histological evaluation, HgS induced enlargement with marked hyperplasia and increase of lymphoid follicles in the spleen. In addition, HgS induced the gene expression of pro-inflammatory cytokines in the spleen and thymus. Our results suggest that insoluble HgS was absorbed by the gastrointestinal tract, accumulated in the spleen and thymus, and thus could affect immune systems.

On self-organized shell formation by bovine carbonic anhydrase II, and soluble protein extracted from regenerated shell.

The soluble protein of hemocytes from diseased shell (HDS) of oyster, Crassostrea gigas, was shown to play a key role in the rapid growth of calcium carbonate crystals. In this study, we compared HDS extracted from regenerated (or diseased) shell with bovine carbonic anhydrase II in terms of their ability to promote the growth of calcium carbonate crystals. On the basis of scanning electron microscopy (SEM) and Fourier transform infrared (FT-IR) analysis, a high growth rate of calcium carbonate crystals was identified under artificial seawater and atmospheric temperature. The function and role of HDS extracted from regenerated shell are discussed at the molecular point as compared to aragonite-specific soluble proteins. Our findings suggest that hemocytes function as a soluble protein, with repeated GX (G: Gly, X: Asp, Asn or Glu) or negative charged amino acid domains binding calcium and specific surface features for catalyzing rapid shell regeneration.

Flavonoids inhibit histamine release and expression of proinflammatory cytokines in mast cells.

Mast cells participate in allergy and inflammation by secreting inflammatory mediators such as histamine and proinflammatory cytokines. Flavonoids are naturally occurring molecules with antioxidant, cytoprotective, and antiinflammatory actions. However, effect of flavonoids on the release of histamine and proinflammatory mediator, and their comparative mechanism of action in mast cells were not well defined. Here, we compared the effect of six flavonoids (astragalin, fisetin, kaempferol, myricetin, quercetin, and rutin) on the mast cell-mediated allergic inflammation. Fisetin, kaempferol, myricetin, quercetin, and rutin inhibited IgE or phorbol-12-myristate 13-acetate and calcium ionophore A23187 (PMACI)-mediated histamine release in RBL-2H3 cells. These five flavonoids also inhibited elevation of intracellular calcium. Gene expressions and secretion of proinflammatory cytokines such as tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-1beta, IL-6, and IL-8 were assessed in PMACI-stimulated human mast cells (HMC-1). Fisetin, quercetin, and rutin decreased gene expression and production of all the proinflammatory cytokines after PMACI stimulation. Myricetin attenuated TNF-alpha and IL-6 but not IL-1beta and IL-8. Fisetin, myricetin, and rutin suppressed activation of NF-kappaB indicated by inhibition of nuclear translocation of NF-kappaB, NF-kappaB/DNA binding, and NF-kappaB-dependent gene reporter assay. The pharmacological actions of these flavonoids suggest their potential activity for treatment of allergic inflammatory diseases through the down-regulation of mast cell activation.

Bisphenol A modulates inflammation and proliferation pathway in human endometrial stromal cells by inducing oxidative stress.

Bisphenol A (BPA) has been implicated in altered human reproductive function. The oxidative stress or change of inflammatory signaling may appear a key factor in the biological changes of the human endometrium. Using MTT assay we assessed BPA mediated modulation of oxidative stress and inflammation responses in human endometrial stromal cells (ESCs). According to the results, reactive oxygen species (ROS) generation was highest upon exposure to 1000 pmol BPA. Increased mitogen-activated protein kinase (MAPK) and nuclear factor-κB (NF-κB) were demonstrated. Gene expression and release of inflammatory cytokines were increased. Upon BPA exposure, elevated estrogen receptor (ER)-α expression levels in ESCs correlated with changes in oxidative stress, inflammatory gene expression and signal changes in cellular proliferation signaling. These findings support that BPA induces oxidative stress and activates inflammatory signals in cultured ESCs via ER-α. Together, this result may provide insight into the association between BPA exposure and endometrium-related disorders.

Assessment of the effects of prostaglandins on myometrial and leiomyoma cells in vitro through microRNA profiling.

It is well known that prostaglandin (PG) E2 and PGF2α are secreted in copious amounts from the menstruating uterus. The aim of the present study was to determine whether PGs affect the growth of uterine leiomyomas (ULs) to the same extent as estrogen or progesterone (P4). The present study evaluated the expression of eight microRNAs (miRNAs) by reverse transcription­quantitative polymerase chain reaction (RT­qPCR) through treatment with estradiol (E2), P4, PGE2, PGF2α and each antagonist or cyclooxygenase­2 (COX­2) inhibitor of cultured leiomyoma and myometrial cells (LC and MC, respectively). The eight miRNAs were divided into two groups according to their primary biological action, namely apoptosis­regulating miRNAs (let­7a, miR­21, miR­26a and miR­200a) and inflammation­regulating miRNAs (miR­29b, miR­93, miR­106b and miR­100b). PGE2 induced significantly higher expression of the 3 anti­apoptotic miRs, let­7a, miR­16a and miR­200a, in LC when compared with the non­treated control or E2. PGE2 significantly promoted a greater expression of let­7a and miR­26a in LC when compared with P4. Overall, PGE2 exerted the highest anti­apoptotic and anti­inflammatory effect in LC, which was comparable with E2. It was not observed among the inflammation­regulating miRNAs in LC. PGF2α did not exert effects as prominent as those of PGE2. In MC, PGs and sex steroids exerted no similar effects on MC compared with LC. The present study demonstrated that PGE2 levels during menstruation may affect the growth of preexisting ULs without affecting the normal myometrium. Therefore, the control of secretion of PGs from the menstruating uterus or the administration of antagonists may be an alternative therapy for inhibiting the growth of ULs.