scholarly journals Changes in Metabolic Profiles of Human Oral Cells by Benzylidene Ascorbates and Eugenol

Medicines ◽  
2018 ◽  
Vol 5 (4) ◽  
pp. 116 ◽  
Author(s):  
Hiroshi Sakagami ◽  
Masahiro Sugimoto ◽  
Yumiko Kanda ◽  
Yukio Murakami ◽  
Osamu Amano ◽  
...  
Keyword(s):  
Early Stage ◽  
Tca Cycle ◽  
Viable Cell ◽  
Cell Number ◽  
Sodium Ascorbate ◽  
Viable Cell Number ◽  
Gingival Fibroblast ◽  
The Tca Cycle ◽  
Oral Cells

Background: Sodium-5,6-benzylidene-L-ascorbate (SBA), and its component units, benzaldehyde (BA) and sodium ascorbate (SA), are known to exert antitumor activity, while eugenol exerts anti-inflammatory activity. To narrow down their intracellular targets, metabolomic analysis was performed. Methods: Viable cell number was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. Fine cell structures were observed under transmission electron microscope. Cellular metabolites were extracted with methanol and subjected to capillary electrophoresis-mass spectrometry (CE-MS) for quantification of intracellular metabolites. Results: SBA was cleaved into BA and SA under acidic condition. Among these three compounds, BA showed the highest-tumor specificity in vitro against human oral squamous cell carcinoma (OSCC) cell line. BA did not induce the vacuolization in HSC-2 OSCC cells, and its cytotoxicity was not inhibited by catalase, in contrast to SBA and SA. Only BA suppressed the tricarboxylic acid (TCA) cycle at early stage of cytotoxicity induction. Eugenol more rapidly induced the vacuolization and suppressed the TCA cycle in three human normal oral cells (gingival fibroblast, periodontal ligament fibroblast, pulp cell). Neither BA nor eugenol affected the ATP utilization, further supporting that they do not induce apoptosis. Conclusions: The present study demonstrated for the first time that both BA and eugenol suppressed the TCA cycle in tumor cells and normal cells, respectively. It is crucial to design methodology that enhances the antitumor potential of BA and reduces the cytotoxicity of eugenol to allow for safe clinical application.

scholarly journals An Experimentally Defined Hypoxia Gene Signature in Glioblastoma and Its Modulation by Metformin

Biology ◽  
2020 ◽  
Vol 9 (9) ◽  
pp. 264
Author(s):  
Marta Calvo Tardón ◽  
Eliana Marinari ◽  
Denis Migliorini ◽  
Viviane Bes ◽  
Stoyan Tankov ◽  
...  
Keyword(s):  
Cell Lines ◽  
In Silico ◽  
Chemotherapy Resistance ◽  
Viable Cell ◽  
Gene Signature ◽  
Cell Number ◽  
Viable Cell Number ◽  
In Silico Analyses

Glioblastoma multiforme (GBM) is the most common and aggressive primary brain tumor, characterized by a high degree of intertumoral heterogeneity. However, a common feature of the GBM microenvironment is hypoxia, which can promote radio- and chemotherapy resistance, immunosuppression, angiogenesis, and stemness. We experimentally defined common GBM adaptations to physiologically relevant oxygen gradients, and we assessed their modulation by the metabolic drug metformin. We directly exposed human GBM cell lines to hypoxia (1% O2) and to physioxia (5% O2). We then performed transcriptional profiling and compared our in vitro findings to predicted hypoxic areas in vivo using in silico analyses. We observed a heterogenous hypoxia response, but also a common gene signature that was induced by a physiologically relevant change in oxygenation from 5% O2 to 1% O2. In silico analyses showed that this hypoxia signature was highly correlated with a perinecrotic localization in GBM tumors, expression of certain glycolytic and immune-related genes, and poor prognosis of GBM patients. Metformin treatment of GBM cell lines under hypoxia and physioxia reduced viable cell number, oxygen consumption rate, and partially reversed the hypoxia gene signature, supporting further exploration of targeting tumor metabolism as a treatment component for hypoxic GBM.

scholarly journals 1α,25-Dihydroxycholecalciferol and a human myeloid leukaemia cell line (HL-60)

1982 ◽  
Vol 204 (3) ◽  
pp. 713-719 ◽  
Author(s):  
H Tanaka ◽  
E Abe ◽  
C Miyaura ◽  
T Kuribayashi ◽  
K Konno ◽  
...  
Keyword(s):  
Myeloid Leukaemia ◽  
Active Form ◽  
Viable Cell ◽  
Cell Number ◽  
Viable Cell Number ◽  
Hormone Action ◽  
Leukaemia Cell ◽  
Leukaemia Cell Line ◽  
Derivatives Of

Human promyelocytic leukaemia cells (HL-60) can be induced to differentiate into mature granulocytes in vitro by 1 alpha,25-dihydroxycholecalciferol [1 alpha,25(OH)2D3], the active form of cholecalciferol. The differentiation-associated properties, such as phagocytosis and C3 rosette formation, were induced by as little as 0.12 nM-1 alpha,25(OH)2D3, and, at 12 nM, about half of the cells exhibited differentiation on day 3 of incubation. Concomitantly the viable cell number was decreased to less than half of the control. Among various derivatives of cholecalciferol examined, 1 alpha,25(OH)2D3 and 1 alpha,24R-dihydroxycholecalciferol were the most potent in inducing differentiation, followed successively by 1 alpha,24S-dihydroxycholecalciferol, 1 alpha-hydroxycholecalciferol, 25-hydroxycholecalciferol and 24R,25-dihydroxycholecalciferol. A cytosol protein specifically bound to 1 alpha,25 (OH)2D3 was found in HL-60 cells. Its physical properties closely resembled those found in such target tissues as intestine and parathyroid glands. 1 alpha,25(OH)2D3 bound to the cytosol receptor was transferred quantitatively to the chromatin fraction. The specificity of various derivatives of cholecalciferol in inducing differentiation was well correlated with that of their association with the cytosol receptor. These results are compatible with the hypothesis that the active form of cholecalciferol induces differentiation of human myeloid leukaemia cells by a mechanism similar to that proposed for the classical concept of steroid hormone action.

scholarly journals Troglitazone suppresses glutamine metabolism through a PPAR-independent mechanism

2015 ◽  
Vol 396 (8) ◽  
pp. 937-947
Author(s):  
Miriam R. Reynolds ◽  
Brian F. Clem
Keyword(s):  
Cell Growth ◽  
Tumor Cell ◽  
Tca Cycle ◽  
Viable Cell ◽  
Cell Number ◽  
Glutamine Metabolism ◽  
Viable Cell Number ◽  
Tumor Cell Growth ◽  
Pparγ Agonist ◽  
Glutamine Uptake

Abstract Enhanced glutamine metabolism is required for tumor cell growth and survival, which suggests that agents targeting glutaminolysis may have utility within anti-cancer therapies. Troglitazone, a PPARγ agonist, exhibits significant anti-tumor activity and can alter glutamine metabolism in multiple cell types. Therefore, we examined whether troglitazone would disrupt glutamine metabolism in tumor cells and whether its action was reliant on PPARγ activity. We found that troglitazone treatment suppressed glutamine uptake and the expression of the glutamine transporter, ASCT2, and glutaminase. In addition, troglitazone reduced 13C-glutamine incorporation into the TCA cycle, decreased [ATP], and resulted in an increase in reactive oxygen species (ROS). Further, troglitazone treatment decreased tumor cell growth, which was partially rescued with the addition of the TCA-intermediate, α-ketoglutarate, or the antioxidant N-acetylcysteine. Importantly, troglitazone’s effects on glutamine uptake or viable cell number were found to be PPARγ-independent. In contrast, troglitazone caused a decrease in c-Myc levels, while the proteasomal inhibitor, MG132, rescued c-Myc, ASCT2 and GLS1 expression, as well as glutamine uptake and cell number. Lastly, combinatorial treatment of troglitazone and metformin resulted in a synergistic decrease in cell number. Therefore, characterizing new anti-tumor properties of previously approved FDA therapies supports the potential for repurposing of these agents.

scholarly journals Effect of Cell Culture Medium on the Proliferation and Stemness of CD24-/CD44+ Human Breast Cancer Stem Cells

2021 ◽  
Vol 13 (4) ◽  
pp. 355-63
Author(s):  
Yalista Fatia Nadia ◽  
Resda Akhra Syahrani ◽  
Sekar Arumsari ◽  
Mohamad Sadikin ◽  
Septelia Inawati Wanandi
Keyword(s):  
Breast Cancer ◽  
Stem Cells ◽  
Cancer Stem Cells ◽  
Human Breast Cancer ◽  
Viable Cell ◽  
Cell Number ◽  
Viable Cell Number ◽  
Breast Cancer Stem Cells ◽  
Human Breast

BACKGROUND: Cancer stem cells (CSCs) is defined as tumor initiating cells within tumor that maintain stemness properties and tumorigenicity. Extracellular pH of CSCs in in vitro condition is important for supporting cell proliferation which may also regulate the expression of stemness markers such as OCT4. This work aimed to examine the effect of cell culture media on the proliferation and stemness of human breast cancer stem cells (BCSCs).METHODS: Human CD24-/CD44+ BCSCs were grown in Dulbecco's Modified Eagle Medium/F-12 (DMEM/F-12) with 15mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), without HEPES and adjusted to pH 7.4, or without HEPES but pH was not adjusted. BCSCs were grown under standard conditions for various days. Viable cell number was measured using trypan blue exclusion, whereas proliferation rate using MTS assay. OCT4 mRNA and protein were analyzed using quantitative real time PCR (qRT-PCR) and Western Blot assay, respectively. In vitro tumorigenic activity was determined using mammosphere formation unit (MFU) assay.RESULTS: Our results showed a higher viable cell number and proliferation of BCSCs in DMEM/F-12 HEPES (-) compared to HEPES (+) medium until 4 day incubation. OCT4 mRNA and protein level, as well as MFU of BCSCs were significantly higher in HEPES (-) compared to HEPES (+) medium on day 2.CONCLUSION: DMEM/F-12 medium without HEPES facilitates CD24-/CD44+ BCSCs to have higher proliferation and stemness on day 2 incubation compared to those with HEPES.KEYWORDS: breast cancer, cancer stem cell, OCT4, stemness, proliferation

scholarly journals Application of a roller conveyor type plasma disinfection device with fungus-contaminated citrus fruits

AMB Express ◽  
2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Akikazu Sakudo ◽  
Yoshihito Yagyu
Keyword(s):  
Treatment Time ◽  
Citrus Fruit ◽  
Viable Cell ◽  
Cell Number ◽  
Viable Cell Number ◽  
Citrus Fruits ◽  
Decimal Reduction Time ◽  
Gas Plasma ◽  
Plasma Device ◽  
Time Required

AbstractEfficient methods to achieve the safe decontamination of agricultural products are needed. Here, we investigated the decontamination of citrus fruits to test the antifungal potential of a novel non-thermal gas plasma apparatus, termed a roller conveyer plasma instrument. This instrument generates an atmospheric pressure dielectric barrier discharge (APDBP) plasma on a set of rollers. Penicillium venetum was spotted onto the surface of the fruit or pericarps, as well as an aluminium plate to act as a control, before performing the plasma treatment. The results showed that viable cell number of P. venetum decreased with a decimal reduction time (D value or estimated treatment time required to reduce viable cell number by 90%) of 0.967 min on the aluminium plate, 2.90 min and 1.88 min on the pericarps of ‘Kiyomi’ (Citrus unshiu × C. sinensis) and ‘Kawano-natsudaidai’ (C. natsudaidai) respectively, and 2.42 min on the surface of ‘Unshu-mikan’ (C. unshiu). These findings confirmed a fungicidal effect of the plasma not only on an abiotic surface (aluminium plate) but also on a biotic surface (citrus fruit). Further development of the instrument by combining sorting systems with the plasma device promises an efficient means of disinfecting citrus fruits during food processing.

scholarly journals INVESTIGATION ON QUALITY PROPERTIES OF TRADITIONAL BULK BREAD COVERED WITH PROBIOTICS AND SOYBEAN OIL EDIBLE COATING

2020 ◽  
Vol 49 (2) ◽  
pp. 144-153
Author(s):  
R. Amiri Qandashtant ◽  
E. Ataye Salehi ◽  
A. Mohamadi Sani ◽  
M. Mehraban Sangatash ◽  
O. Safari
Keyword(s):  
Soybean Oil ◽  
Whey Protein ◽  
Corn Starch ◽  
Lactobacillus Rhamnosus ◽  
Methyl Cellulose ◽  
Viable Cell ◽  
Cell Number ◽  
Whey Protein Concentrate ◽  
Viable Cell Number ◽  
Quality Properties

Probiotic food products are available at the supermarket commercially, but probiotic bakery products are much less in evidence. In the present study, methyl cellulose (2%), whey protein concentrate (2%), corn starch (1%), and soybean oil at 2, 4, and 6% were used for coating layer on the bulked bread surface, and then the quality properties were studied. The results showed that Lactobacillus rhamnosus GG, as probiotic component of the coating, immobilized in corn starch, whey protein, and methyl cellulose films had enhanced viability throughout shelf-life. The probiotics remained viable for 4 days, maintaining high viable cell number levels. Adding soybean oil at 6% concentration enhanced texture, sensory properties, and image index during storage.

scholarly journals Metabolomic profiling of tumor-infiltrating macrophages during tumor growth

2020 ◽  
Vol 69 (11) ◽  
pp. 2357-2369
Author(s):  
Naoki Umemura ◽  
Masahiro Sugimoto ◽  
Yusuke Kitoh ◽  
Masanao Saio ◽  
Hiroshi Sakagami
Keyword(s):  
T Cells ◽  
Tumor Growth ◽  
Cd8 T Cells ◽  
Early Stage ◽  
Suppressor Cells ◽  
Tca Cycle ◽  
Tumor Stage ◽  
Cell Number ◽  
Tumor Bearing ◽  
Intracellular Metabolism

Abstract Myeloid-derived suppressor cells (MDSCs) and tumor-associated macrophages (TAMs) are both key immunosuppressive cells that contribute to tumor growth. Metabolism and immunity of tumors depend on the tumor microenvironment (TME). However, the intracellular metabolism of MDSCs and TAMs during tumor growth remains unclear. Here, we characterized CD11b+ cells isolated from a tumor-bearing mouse model to compare intratumoral TAMs and intrasplenic MDSCs. Intratumoral CD11b+ cells and intrasplenic CD11b+ cells were isolated from tumor-bearing mice at early and late stages (14 and 28 days post-cell transplantation, respectively). The cell number of intrasplenic CD11b+ significantly increased with tumor growth. These cells included neutrophils holding segmented leukocytes or monocytes with an oval nucleus and Gr-1hi IL-4Rαhi cells without immunosuppressive function against CD8 T cells. Thus, these cells were classified as MDSC-like cells (MDSC-LCs). Intratumoral CD11b+ cells included macrophages with a round nucleus and were F4/80hi Gr-1lo IL-4Rαhi cells. Early stage intratumoral CD11b+ cells inhibited CD8 T cells via TNFα. Thus, this cell population was classified as TAMs. Metabolomic analyses of intratumoral TAMs and MDSC-LCs during tumor growth were conducted. Metabolic profiles of intratumoral TAMs showed larger changes in various metabolic pathways, e.g., glycolysis, TCA cycle, and glutamic acid pathways, during tumor growth compared with MDSL-LCs. Our findings demonstrated that intratumoral TAMs showed an immunosuppressive capacity from the early tumor stage and underwent intracellular metabolism changes during tumor growth. These results clarify the intracellular metabolism of TAMs during tumor growth and contribute to our understanding of tumor immunity.

Mcl-1 in Transgenic Mice Promotes Survival in a Spectrum of Hematopoietic Cell Types and Immortalization in the Myeloid Lineage

Blood ◽  
1998 ◽  
Vol 92 (9) ◽  
pp. 3226-3239 ◽  
Author(s):  
Ping Zhou ◽  
Liping Qian ◽  
Christine K. Bieszczad ◽  
Randolph Noelle ◽  
Michael Binder ◽  
...  
Keyword(s):  
Myeloid Cells ◽  
Myeloid Cell ◽  
Cell Types ◽  
Cell Number ◽  
Hematopoietic Cell ◽  
Viable Cell Number ◽  
Wide Range ◽  
Transgenic Cells

Abstract Mcl-1 is a member of the Bcl-2 family that is expressed in early monocyte differentiation and that can promote viability on transfection into immature myeloid cells. However, the effects of Mcl-1 are generally short lived compared with those of Bcl-2 and are not obvious in some transfectants. To further explore the effects of this gene, mice were produced that expressed Mcl-1 as a transgene in hematolymphoid tissues. The Mcl-1 transgene was found to cause moderate viability enhancement in a wide range of hematopoietic cell types, including lymphoid (B and T) as well as myeloid cells at both immature and mature stages of differentiation. However, enhanced hematopoietic capacity in transgenic bone marrow and spleen was not reflected in any change in pool sizes in the peripheral blood. In addition, among transgenic cells, mature T cells remained long lived compared with B cells and macrophages could live longer than either of these. Interestingly, when hematopoietic cells were maintained in tissue culture in the presence of interleukin-3, Mcl-1 enhanced the probability of outgrowth of continuously proliferating myeloid cell lines. Thus, Mcl-1 transgenic cells remained subject to normal in vivo homeostatic mechanisms controlling viable cell number, but these constraints could be overridden under specific conditions in vitro. Within the organism, Bcl-2 family members may act at “viability gates” along the differentiation continuum, functioning as part of a system for controlled hematopoietic cell amplification. Enforced expression of even a moderate viability-promoting member of this family such as Mcl-1, within a conducive intra- and extracellular environment in isolation from normal homeostatic constraints, can substantially increase the probability of cell immortalization. © 1998 by The American Society of Hematology.

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