scholarly journals Whole Peptidoglycan Extracts from theLactobacillus paracaseisubsp.paracaseiM5 Strain Exert Anticancer ActivityIn Vitro

2018 ◽  
Vol 2018 ◽  
pp. 1-11 ◽  
Author(s):  
Shumei Wang ◽  
Xue Han ◽  
Lanwei Zhang ◽  
Yingchun Zhang ◽  
Hongbo Li ◽  
...  
Keyword(s):  
Lactobacillus Paracasei ◽  
Molar Ratio ◽  
Dependent Manner ◽  
Apoptotic Pathway ◽  
Trypan Blue Exclusion ◽  
Anticancer Effects ◽  
Transmission Electron ◽  
Sds Page ◽  
Dose Dependent ◽  
Ht 29

TheLactobacillus paracaseisubsp. paracaseiM5 strain exerted potential anticancer activity through the cell wall. In this study, whole peptidoglycan (WPG) was extracted from theLactobacillus paracaseisubsp. paracaseiM5 strain and was evaluated for anticancer effects as well as its properties. SDS-PAGE analysis confirmed the presence of WPG with dominant bands of approximately 14.4 kDa. Further analysis revealed that the amino acids present in the WPG consisted of alanine, glycine, glutamic acid, and lysine in a molar ratio of approximately 8 : 5 : 3 : 3.5. In addition, the cell viability of HT-29 cells with WPG addition was investigated with methyl thiazolyl tetrazolium (MTT) and trypan blue exclusion (TBE) assays, and cell apoptosis was analyzed with a transmission electron microscope, flow cytometry, and semiquantitative RT-PCR. These results showed that WPG exerted cytotoxic effects on colon cancer HT-29 cells in a dose-dependent manner and upregulated proapoptotic genes, while downregulating antiapoptotic genes. The gene expression study definitively revealed that WPG induced a mitochondria-mediated apoptotic pathway.

scholarly journals Oxymatrine Synergistically Enhances Doxorubicin Anticancer Effects in Colorectal Cancer

2021 ◽  
Vol 12 ◽  
Author(s):  
Di Pan ◽  
Wen Zhang ◽  
Nenling Zhang ◽  
Yini Xu ◽  
Yi Chen ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Western Blotting ◽  
Combination Group ◽  
Ros Generation ◽  
Synergistic Activity ◽  
Dependent Manner ◽  
Anticancer Effects ◽  
Sw620 Cells ◽  
Dose Dependent ◽  
Ht 29

The combination of chemotherapy with natural products is a common strategy to enhance anticancer effects while alleviating the dose-dependent adverse effects of cancer treatment. Oxymatrine (OMT) has been extensively reported as having anticancer activity. Doxorubicin (DOX) is a chemotherapeutic DNA-damaging agent used for the treatment of carcinoma. In this study, we investigated whether synergistic effects exist with the combination treatment with OMT and DOX using human colorectal cancer cell (CRC) lines and the potential mechanisms involved in in vitro and in vivo activities. The MTT and colony formation assay results showed that compared to either OMT or DOX monotherapy, the combination of OMT + DOX markedly inhibited the growth of HT-29 and SW620 cells. Wound healing assays showed significant inhibition of cell migration with co-treatment, supported by the change in E-cadherin and N-cadherin expressions in Western blotting. Furthermore, flow cytometry analysis revealed that OMT + DOX co-treatment enhanced cell apoptosis as a result of ROS generation, whereas NAC attenuated OMT + DOX–induced apoptosis. Similarly, the apoptosis-related proteins (cleaved caspase-3, cleaved caspase-9, and the ratio of Bax/Bcl-2) were determined by Western blotting, which showed that the expressions of these markers were notably increased in the co-treatment group. Furthermore, co-administration of a low dose of DOX and OMT inhibited xenograft tumor growth in a dose-dependent manner. TUNEL assay and Ki67 staining images indicated more apoptosis and less proliferation occurred in OMT plus DOX-treated xenograft tumors. Meanwhile, the combination strategy decreased cardiotoxicity, which is the most serious side effect of DOX. RNA sequencing was performed to explore the precise molecular alterations involved in the combination group. Among the numerous differentially expressed genes, downregulated FHL-2 and upregulated cleaved SPTAN1 were validated in both mRNA and protein levels of HT-29 and SW620 cells. These two proteins might play a pivotal role involving in OMT + DOX synergistic activity. Overall, OMT in combination with DOX presented an outstanding synergistic antitumor effect, indicating that this beneficial combination may offer a potential therapy for CRC patients.

scholarly journals Synthesis, Characterization, and Optimization of Green Silver Nanoparticles Using Neopestalotiopsis clavispora and Evaluation of Its Antibacterial, Antibiofilm, and Genotoxic Effects

2021 ◽  
Vol 5 (3) ◽  
pp. 109-122
Author(s):  
Tuğba Kahraman ◽  
Safiye Elif Korcan ◽  
Recep Liman ◽  
İbrahim Hakkı Ciğerci ◽  
Yaser Acikbas ◽  
...  
Keyword(s):  
Silver Nanoparticles ◽  
Nuclear Ribosomal Dna ◽  
Diffusion Method ◽  
Dependent Manner ◽  
Transmission Electron ◽  
Infrared Spectrophotometer ◽  
Average Size ◽  
Novel Applications ◽  
Dose Dependent

Abstract Silver nanoparticles (AgNPs) have been used in a variety of biomedical applications in the last two decades, including antimicrobial, anti-inflammatory, and anticancer treatments. The present study highlights the extracellular synthesis of silver nanoparticles AgNPs using Neopestalotiopsis clavispora MH244410.1 and its antibacterial, antibiofilm, and genotoxic properties. Locally isolated N. clavispora MH244410.1 was identified by Internal transcribed spacer (ITS) sequences of nuclear ribosomal DNA. Optimization of synthesized AgNPs was performed by using various parameters (pH (2, 4, 7, 9 and 12), temperature (25, 35 and 45 °C), and substrate concentration (0.05, 0.1, 0.15, 0.2 and 0.25 mM)). After 72 hours of incubation in dark conditions, the best condition for the biosynthesis of AgNPs was determined as 0.25 mM metal concentration at pH 12 and 35 °C. Fungal synthesized AgNPs were characterized via spectroscopic and microscopic techniques such as Fouirer Transform Infrared Spectrophotometer (FTIR), UV-Visible Spectroscopy, and Transmission Electron Microscopy (TEM). The average size of the AgNPs was determined less than 60 nm using the TEM and Zetasizer measurement system (measured in purity water suspension). The characteristic peak of AgNPs was observed at ~414 nm from UV-Vis results. Antibacterial and genotoxic activity of synthesized AgNPs (0.1, 1, and 10 ppm) were also determined by using the agar well diffusion method and in vivo Somatic Mutation and Recombination Test (SMART) in Drosophila melanogaster. AgNPs exhibited potential antimicrobial activity against all the tested bacteria (Bacillus subtilis, Staphylococcus aureus, and Pseudomonas aeruginosa) except Escherichia coli in a dose-dependent manner. AgNPs did not induce genotoxicity in the Drosophila SMART assay. 79.33, 65.47, and 41.95% inhibition of biofilms formed by P. aeruginosa were observed at 10, 1, and 0.1 ppm of AgNPs, respectively. The overall results indicate that N. clavispora MH244410.1 is a good candidate for novel applications in biomedical research.

scholarly journals Antioxidant, Antibacterial, and Cytoprotective Activity of Agathi Leaf Protein

2014 ◽  
Vol 2014 ◽  
pp. 1-8 ◽  
Author(s):  
A. S. Zarena ◽  
Shubha Gopal ◽  
R. Vineeth
Keyword(s):  
Safety Assessment ◽  
Crude Protein ◽  
Leaf Protein ◽  
Dependent Manner ◽  
Cytoprotective Activity ◽  
Sesbania Grandiflora ◽  
Rp Hplc ◽  
Maximum Inhibition ◽  
Sds Page ◽  
Dose Dependent

In the present study a protein termed agathi leaf protein (ALP) fromSesbania grandiflora Linn. (agathi) leaves was isolated after successive precipitation with 65% ammonium sulphate followed by purification on Sephadex G 75. The column chromatography of the crude protein resulted in four peaks of which Peak I (P I) showed maximum inhibition activity against hydroxyl radical. SDS-PAGE analysis of P I indicated that the molecular weight of the protein is≈29 kDa. The purity of the protein was 98.4% as determined by RP-HPLC and showed a single peak with a retention time of 19.9 min. ALP was able to reduce oxidative damage by scavenging lipid peroxidation against erythrocyte ghost (85.50 ± 6.25%), linolenic acid (87.67 ± 3.14%) at 4.33 μM, ABTS anion (88 ± 3.22%), and DNA damage (83 ± 4.20%) at 3.44 μM in a dose-dependent manner. The purified protein offered significant protection to lymphocyte (72% at 30 min) induced damage by t-BOOH. In addition, ALP showed strong antibacterial activity againstPseudomonas aeruginosa(20 ± 3.64 mm) andStaphylococcus aureus(19 ± 1.53 mm) at 200 μg/mL. The safety assessment showed that ALP does not induce cytotoxicity towards human lymphocyte at the tested concentration of 0.8 mg/mL.

scholarly journals Inhibitory effect of gamma irradiation against Cucumber green mottle mosaic virus

2017 ◽  
Vol 53 (No. 4) ◽  
pp. 201-207 ◽  
Author(s):  
Hong Jin-Sung ◽  
Jeong Mi-Ae ◽  
Jeong Rae-Dong
Keyword(s):  
Gamma Irradiation ◽  
Mosaic Virus ◽  
Fungal Pathogens ◽  
Inhibitory Effect ◽  
Plant Viruses ◽  
Dependent Manner ◽  
Genomic Rna ◽  
Transmission Electron ◽  
Virion Structure ◽  
Dose Dependent

Gamma irradiation has been shown to be an effective method of controlling plant bacterial and fungal pathogens, but data on its effect against plant viruses is limited. A mechanism for the inactivation of plant viruses by gamma irradiation has not been proposed. Gamma irradiation was evaluated for the inactivation of Cucumber green mottle mosaic virus (CGMMV) in Nicotiana tabacum plants. CGMMV infectivity decreased gradually in a dose-dependent manner, and the virus was completely inactivated at over 40 kGy. Transmission electron microscopy revealed that gamma irradiation disrupts the virion structure and degrades viral protein and genomic RNA, suggesting that damage to viral constituents is the mechanism by which gamma irradiation inactivates the virus.

Alpha mangostin Inhibits Hepatic Stellate Cells Activation Through TGF-β/Smad and Akt Signaling Pathways: An in vitro Study in LX2

Drug Research ◽  
2017 ◽  
Vol 68 (03) ◽  
pp. 153-158 ◽  
Author(s):  
Rahmaniah Rahmaniah ◽  
Yuyuntia Yuyuntia ◽  
Vivian Soetikno ◽  
Wawaimuli Arozal ◽  
Radiana Antarianto ◽  
...  
Keyword(s):  
Signaling Pathways ◽  
Hepatic Stellate Cells ◽  
Stellate Cells ◽  
Akt Signaling ◽  
Dependent Manner ◽  
Trypan Blue Exclusion ◽  
Ki 67 ◽  
Trypan Blue Exclusion Method ◽  
Dose Dependent

Abstract Background Alpha mangostin has been reported to have activity for the treatment of liver fibrosis in the rats. However, the mechanisms of action are poorly understood. This study was aimed to investigate the effect of alpha mangostin on hepatic stellate cells (HSC) activation and proliferation through TGF-β/Smad and Akt signaling pathways. Methods Immortalized HSC, LX2 cells, were incubated with TGF-β with or without alpha mangostin (5 or 10 μM). Sorafenib 10 µM was used as positive control. LX2 viability was counted using trypan blue exclusion method. The effect of alpha mangostin on TGF-β concentrations, and the expressions of proliferation and fibrogenic markers were evaluated. Results Alpha mangostin treatment resulted in a reduced proliferation of HSC, decreased Ki-67 and p-Akt expressions. These findings were followed with decreased concentrations of TGF-β in the medium of cells treated with alpha mangostin, decreased expressions of COL1A1, TIMP1, PAI1, α-SMA, and p-Smad3 as fibrogenic markers. These effects were shown to be dose-dependent. Conclusions Alpha mangostin inhibits hepatic stellate cells proliferation and activation through TGF-β/Smad and Akt signaling pathways in dose dependent manner.

Effect of Estradiol on miR-21& miR-155 Expression in promyelocytic leukemia-derived cell line NB4

2020 ◽  
Author(s):  
Robab Naghibzadeh ◽  
Narges Obeidi ◽  
Alireza Farsinejd ◽  
Gholamreza Khamisipour ◽  
Masoud TohidFar
Keyword(s):  
Effective Dose ◽  
Cell Line ◽  
Myeloid Leukemia ◽  
Cellular Proliferation ◽  
Promyelocytic Leukemia ◽  
Dependent Manner ◽  
Trypan Blue Exclusion ◽  
Nb4 Cells ◽  
Dose Dependent ◽  
Mtt Assays

Background: Due to the estrogen participation in modulating the proliferation and commitment of stem cells and the effects of miR-21 and miR-155 expression on reduced proliferation and colony formation of acute myeloid leukemia (AML), the aim of the present study was to evaluate the effect of estradiol on expression of miR-21 and miR-155 in the NB4 cell line, as an acute promyelocytic leukemia (APL). Materials and Methods: In the present experiment, NB4 cells were treated with different quantities of estradiol (5, 25, 50, 75, 100, 150, 200, 250 μg/ml) and vehicle control for 24 and 48 hours. Viability, apoptosis, and cellular proliferation were estimated by trypan blue exclusion, flow cytometry, and MTT assays, respectively. The level of miR-155 and miR-21 expression was studied using absolute quantitative real-time PCR. Results: Results showed that estradiol in the effective dose (200 μg/ml) led to decreased cellular viability (in a dose dependent manner, P = 0.004) and apoptosis of NB4 cells. In addition, the expressions of miR-155 and miR-21 were significantly and dose-dependently decreased (p<0.05). Conclusion: Estradiol at the effective dose caused apoptosis in NB4 cell line. This substance can be used as a drug for the treatment of APL. However, further assessments are needed to support the effectiveness of estradiol in the treatment of APL.

L(+)-Lactate Binding to a Protein in Rat Skeletal Muscle Plasma Membranes

1995 ◽  
Vol 20 (1) ◽  
pp. 112-124 ◽  
Author(s):  
Karl J. A. McCullagh ◽  
Arend Bonen
Keyword(s):  
Skeletal Muscle ◽  
Plasma Membranes ◽  
Mass Range ◽  
Dependent Manner ◽  
Rat Skeletal Muscle ◽  
Lactate Transport ◽  
Sds Page ◽  
Lactate And Pyruvate ◽  
Potential Inhibitors ◽  
Dose Dependent

Biochemical studies were conducted to determine the location of a putative lactate transport protein in rat skeletal muscle plasma membranes (PM). PM (50-100 μg protein) were incubated with [U-14C] L(+)-lactate, in the presence or absence of unlabeled monocarboxylates or potential inhibitors, after which proteins were separated by SDS-PAGE. Gel slices (2 mm) were cut and analyzed for14C. [U-14C] L(+)-lactate was bound to plasma membranes in the 30 to 40 kDa molecular mass range. Binding of [U-14C] L(+)-lactate was inhibited by N-ethylmaleimide, unlabeled L-lactate and pyruvate, and in a dose dependent manner by α-cyano-4-hydroxycinnamate (r = 0.995), but not by cytochalasin-B. The inhibition of [U-14C] L(+)-lactate binding was similar to the inhibition of lactate transport. Therefore the transport of L(+)-lactate across skeletal muscle plasma membranes involves a polypeptide of 30 to 40 kDa. Key words: transport, affinity labeling

scholarly journals Measurement of tissue transglutaminase activity in a permeabilized cell system: its regulation by Ca2+ and nucleotides

1996 ◽  
Vol 313 (3) ◽  
pp. 803-808 ◽  
Author(s):  
Peter A. SMETHURST ◽  
Martin GRIFFIN
Keyword(s):  
Tissue Transglutaminase ◽  
Cell System ◽  
Dependent Manner ◽  
Human Endothelial Cells ◽  
Permeabilized Cell ◽  
Protein Substrates ◽  
Endogenous Protein ◽  
Sds Page ◽  
Competitive Inhibitors ◽  
Dose Dependent

Electropermeabilized human endothelial cells (ECV-304) were used to study the regulation of tissue transglutaminase (tTGase) activity in the intracellular environment. An ELSA (enzyme-linked sorbent assay) plate assay was developed for intracellular tTGase activity, using the incorporation of a biotinylated primary amine, 5-{[(N-biotinoylamino)hexanoyl]amino}pentylamine (biotin-x-cadaverine; BTC), into endogenous protein substrates of tTGase. This incorporation process was inhibited by competitive inhibitors of tTGase, cystamine and monodansylcadaverine, in a dose-dependent manner. Over a 30 min period tTGase and its protein substrates did not leak out of the cell, and no incorporation of BTC occurred in unpermeabilized cells, indicating the reaction to be intracellular. In the presence of 10 nM or 10 μM Ca2+, when nucleotides ATP and GTP were added at concentrations mimicking cytosolic levels, tTGase activity was decreased virtually to zero. Only at 100 μM Ca2+, when nucleotides were low or absent was tTGase activity observed. Under these conditions a variety of proteins was labelled by the enzyme, with the major labelling found in a protein of molecular mass around 51 kDa when analysed by SDS/PAGE/Western blotting.

scholarly journals Anticancer Potential of L-Histidine-Capped Silver Nanoparticles against Human Cervical Cancer Cells (SiHA)

Nanomaterials ◽  
2021 ◽  
Vol 11 (11) ◽  
pp. 3154
Author(s):  
Rajmohamed Mohammed Asik ◽  
Chidhambaram Manikkaraja ◽  
Karuppusamy Tamil Surya ◽  
Natarajan Suganthy ◽  
Archunan Priya Aarthy ◽  
...  
Keyword(s):  
Cervical Cancer ◽  
Silver Nanoparticles ◽  
Microscopic Analysis ◽  
Dependent Manner ◽  
Apoptotic Pathway ◽  
Ic50 Value ◽  
Cervical Cancer Cells ◽  
Microscopic Studies ◽  
Human Cervical Cancer Cells ◽  
Dose Dependent

This study reports the synthesis of silver nanoparticles using amino acid L-histidine as a reducing and capping agent as an eco-friendly approach. Fabricated L-histidine-capped silver nanoparticles (L-HAgNPs) were characterized by spectroscopic and microscopic studies. Spherical shaped L-HAgNPs were synthesized with a particle size of 47.43 ± 19.83 nm and zeta potential of −20.5 ± 0.95 mV. Results of the anticancer potential of L-HAgNPs showed antiproliferative effect against SiHa cells in a dose-dependent manner with an IC50 value of 18.25 ± 0.36 µg/mL. Fluorescent microscopic analysis revealed L-HAgNPs induced reactive oxygen species (ROS) mediated mitochondrial dysfunction, leading to activation of apoptotic pathway and DNA damage eventually causing cell death. To conclude, L-HAgNPs can act as promising candidates for cervical cancer therapy.

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