scholarly journals Investigating the Inhibitory Aspects of Metformin/Curcumin Co-Treatment through Convergence of In-Silico and In-Vitro Approaches

2019 ◽  
Author(s):  
Farzaneh Afzali ◽  
Zahra Nayeri ◽  
Zarrin Minuchehr ◽  
Mossa Gardaneh
Keyword(s):  
Treatment Strategy ◽  
In Silico ◽  
Treatment Optimization ◽  
Risk Of Death ◽  
Viability Assay ◽  
Mcf7 Cell Line ◽  
Underlying Mechanisms ◽  
Pathway Analyses ◽  
Functional Mechanisms

ABSTRACTNearly 16% of people with breast cancer (BC) have Diabetes Mellitus type 2 (DM2) and are at a higher risk of death worldwide. Their common regulatory factors and functional mechanisms can be targeted applying multi-target drugs including Metformin (MTFN) and Curcumin (CURC). In this study, we used in-silico approaches to study the potential underlying mechanisms of this co-treatment strategy on BC and DM2 in order to introduce novel therapeutic targets.The total number of 48 shared differentially expressed genes (17 up-regulated and 31 down-regulated) were identified through establishing diseases’ protein-protein network and BC RNA-sequencing expression data. The integration of functional clustering and pathway analyses revealed that the most involved cellular pathways and processes are regard to cells’ proliferation, death, migration, and response to external stimulus. Afterwards, the MTFN/CURC correlation and co-treatment optimization was probed through response surface methodology (RSM) based on MCF7 cell line and confirmed by MDA-MB-231. Combination index calculation by MTT viability assay proved supportive effects on both cell lines. The superior apoptotic potential of co-treatment compared to single treatments was shown on inhibition of MCF7 proliferation and induction of cell death demonstrated by cell body co-staining and flow cytometry as well as gene expression analysis via RT-PCR. Furthermore, wound-healing scratch assay showed that this co-treatment has a slightly higher effect on migration inhibition compared to single treatments.In conclusion, our study used in-silico and in-vitro approaches and introduced a potential regulatory panel between BC and DM2. We also provided a linear model and equation that show the positive relation of drugs’ co-treatment. The proposed co-treatment strategy successfully controlled the biological processes under investigation.

scholarly journals In vitro and in silico anti-dengue activity of compounds obtained from Psidium guajava through bioprospecting

2019 ◽  
Vol 19 (1) ◽  
Author(s):  
Andrea Isabel Trujillo-Correa ◽  
Diana Carolina Quintero-Gil ◽  
Fredyc Diaz-Castillo ◽  
Winston Quiñones ◽  
Sara M. Robledo ◽  
...  
Keyword(s):  
Gallic Acid ◽  
Treatment Strategy ◽  
In Silico ◽  
Antiviral Effect ◽  
Psidium Guajava ◽  
Caribbean Coast ◽  
Ethanolic Extracts ◽  
Pharmacological Potential ◽  
Experimental Strategies

Abstract Background For decades, bioprospecting has proven to be useful for the identification of compounds with pharmacological potential. Considering the great diversity of Colombian plants and the serious worldwide public health problem of dengue—a disease caused by the dengue virus (DENV)—in the present study, we evaluated the anti-DENV effects of 12 ethanolic extracts derived from plants collected in the Colombian Caribbean coast, and 5 fractions and 5 compounds derived from Psidium guajava. Methods The cytotoxicity and antiviral effect of 12 ethanolic extracts derived from plants collected in the Colombian Caribbean coast was evaluated in epithelial VERO cells. Five fractions were obtained by open column chromatography from the ethanolic extract with the highest selectivity index (SI) (derived from P. guajava, SI: 128.2). From the fraction with the highest selectivity (Pg-YP-I-22C, SI: 35.5), five compounds were identified by one- and two-dimensional nuclear magnetic resonance spectroscopy. The antiviral effect in vitro of the fractions and compounds was evaluated by different experimental strategies (Pre- and post-treatment) using non-toxic concentrations calculated by MTT method. The DENV inhibition was evaluated by plate focus assay. The results were analyzed by means of statistical analysis using Student’s t-test. Finally the antiviral effect in Silico was evaluated by molecular docking. Results In vitro evaluation of these compounds showed that three of them (gallic acid, quercetin, and catechin) were promising antivirals as they inhibit the production of infectious viral particles via different experimental strategies, with the best antiviral being catechin (100% inhibition with a pre-treatment strategy and 91.8% with a post-treatment strategy). When testing the interactions of these compounds with the viral envelope protein in silico by docking, only naringin and hesperidin had better scores than the theoretical threshold of − 7.0 kcal/mol (− 8.0 kcal/mol and − 8.2 kcal/mol, respectively). All ligands tested except gallic acid showed higher affinity to the NS5 protein than the theoretical threshold. Conclusion Even though bioprospecting has recently been replaced by more targeted tools for identifying compounds with pharmacological potential, our results show it is still useful for this purpose. Additionally, combining in vitro and in silico evaluations allowed us to identify promising antivirals as well as their possible mechanisms of action.

Evaluation of EZH2 SNPs in cholangiocarcinoma patients.

2012 ◽  
Vol 30 (15_suppl) ◽  
pp. 10611-10611
Author(s):  
Elisa Paolicchi ◽  
Paola Pacetti ◽  
Elisa Giovannetti ◽  
Andrea Mambrini ◽  
Massimo Orlandi ◽  
...  
Keyword(s):  
Clinical Outcome ◽  
In Silico ◽  
Functional Characterization ◽  
Polycomb Group ◽  
Missense Mutations ◽  
Nucleotide Polymorphisms ◽  
Polycomb Group Protein ◽  
Treatment Optimization ◽  
Logrank Test ◽  
Risk Of Death

10611 Background: Cholangiocarcinoma (CCA) is an aggressive tumor arising from biliary tract epithelium.CCA is the second most common primary hepatic malignancy, with a progressive increasing incidence in western countries. Polycomb group protein Enhancer of Zeste homolog 2 (EZH2) is overexpressed in several human carcinomas, including CCA, where EZH2 overexpression is associated with tumor progression. The aim of this study is to evaluate the correlation between candidate EZH2 Single Nucleotide Polymorphisms (SNPs) with clinical outcome in CCA patients. Methods: Genomic DNA was extracted from blood samples of 75 patients [44 male and 31 female, with average age of 62.3 (range, 26-80 years)] affected by hystologically confirmed advanced CCA, treated with the epirubicin-cisplatin-xeloda (ECX) regimen. We performed an in silico characterization to select EZH2 SNPs (rs2302427 C/G, rs6464926 C/T, rs17171119 T/G and rs887569 C/T) from 26 EZH2 SNPs described previously. Genotyping was performed through Taqman PCR. Prognostic value of selected EZH2 SNPs was assesses by correlation with time to progression (TTP) and overall survival (OS). OS and TTP curves were obtained through Kaplan-Meier method, and comparison with survival distribution was evaluated with logrank test. Results: Through specific software (PROMO 3.0, MicroSNiper and Gene Card) we performed an in silico analysis based on functional characterization criteria (transcription factor binding-TFB, miRNA binding and missense mutations). We selected 4 EZH2 SNP alleles showing specific relevance in CCA because of their differential TFB affinity (PPARα/RXRα, E2F-1, Pax-5 and p53). The rs887569 C/T SNP was significantly associated with clinical outcome. The TT genotype predicted a significantly longer OS in CCA patients (TT vs CT-CC p=0.026). Moreover, the TT genotype showed a trend-like association with reduced risk of death (HR=0.59, 95%CI=0.33-1.05, p=0.075), at multivariate analysis. Conclusions: These results suggest the role of rs887569 EZH2 SNP as a possible predictive marker of OS in advanced CCA patients treated with ECX regimen, and offer a potential new tool for treatment optimization.

Naturally Occurring Level of Aflatoxin B1 Injures Human, Canine and Bovine Leukocytes Through ATP Depletion and Caspase Activation

2019 ◽  
Vol 39 (1) ◽  
pp. 30-38 ◽  
Author(s):  
Jalil Mehrzad ◽  
Fatemeh Fazel ◽  
Nazaninzeynam Pouyamehr ◽  
Saman Hosseinkhani ◽  
Hesam Dehghani
Keyword(s):  
Flow Cytometry ◽  
Caspase 3 ◽  
Companion Animals ◽  
Atp Depletion ◽  
Caspase Activation ◽  
Viability Assay ◽  
Naturally Occurring ◽  
Old Persian ◽  
Underlying Mechanisms

Aflatoxin (AF) B1 is a potent hepatotoxic, mutagenic, teratogenic mycotoxin and may cause immune suppression/dysregulation in humans and animals. Toxic effects of AFB1 on key mammalian immune cells (ie, leukocytes) needs to be mechanistically elucidated. In this study, along with the determination of AFB1’s LC50 for certain leukocytes, we analyzed the effect of naturally occurring levels of AFB1 on apoptosis/necrosis of neutrophils, lymphocytes, and monocytes from healthy young humans (20- to 25-year-old male), dogs (1- to 2-year-old Persian/herd breed), and cattle (1- to 2-year-old cattle). Leukocytes were incubated for approximately 24 hours with naturally occurring levels of AFB1 (10 ng/mL). Intracellular adenosine triphosphate (ATP) depletion and caspase-3/7 activity were then determined by luciferase-dependent bioluminescence (BL). Furthermore, the necrotic leukocytes were measured using propidium iodide (PI)-related flow cytometry. A significant decrease (24%-45%, 33.2% ± 2.7%) in intracellular ATP content was observed in AFB1-treated neutrophils, lymphocytes, and monocytes in all studied mammals. Also, with such a low level (10 ng/mL) of AFB1, BL-based caspase-3/7 activity (BL intensity) in all 3 tested mammalian leukocyte lineages was noticeably increased (∼>2-fold). Flow cytometry-based PI staining (for viability assay) of the AFB1-treated leukocytes showed slightly/insignificantly more increase of necrotic (PI+) neutrophils, lymphocytes, and monocytes in human, dogs, and cattle. Even though in vitro LC50s for AFB1’ (∼20,000-40,000 ng/mL) were approximately 2,000 to 4,000 times higher than background, these studies demonstrate leukocytes from human and farm/companion animals are sensitive to naturally occurring levels of AFB1. The observed in vitro ATP depletion and caspase activation in AFB1-exposed leukocytes can partially explain the underlying mechanisms of AFB1-induced immune disorders in mammals.

In vitro and in silico anti-osteoporosis activities and underlying mechanisms of a fructan, ABW90-1, from Achyranthes bidentate

2021 ◽  
pp. 118730
Author(s):  
Tianyu Li ◽  
Xin Hou ◽  
Yihua Huang ◽  
Changsheng Wang ◽  
Haiyun Chen ◽  
...  
Keyword(s):  
In Silico ◽  
Underlying Mechanisms

scholarly journals Structural and functional consequences of succinate dehydrogenase subunit B mutations

2015 ◽  
Vol 22 (3) ◽  
pp. 387-397 ◽  
Author(s):  
E Kim ◽  
E M Rath ◽  
V H M Tsang ◽  
A P Duff ◽  
B G Robinson ◽  
...  
Keyword(s):  
Succinate Dehydrogenase ◽  
In Silico ◽  
Homology Model ◽  
Crystallographic Structure ◽  
Structural Modelling ◽  
Gene Encoding ◽  
Underlying Mechanisms ◽  
Succinate Dehydrogenase Subunit B ◽  
Subunit B

Mitochondrial dysfunction, due to mutations of the gene encoding succinate dehydrogenase (SDH), has been implicated in the development of adrenal phaeochromocytomas, sympathetic and parasympathetic paragangliomas, renal cell carcinomas, gastrointestinal stromal tumours and more recently pituitary tumours. Underlying mechanisms behind germline SDH subunit B (SDHB) mutations and their associated risk of disease are not clear. To investigate genotype–phenotype correlation of SDH subunit B (SDHB) variants, a homology model for humanSDHwas developed from a crystallographic structure.SDHBmutations were mapped, and biochemical effects of these mutations were predictedin silico. Results of structural modelling indicated that many mutations withinSDHBare predicted to cause either failure of functionalSDHBexpression (p.Arg27*, p.Arg90*, c.88delC and c.311delAinsGG), or disruption of the electron path (p.Cys101Tyr, p.Pro197Arg and p.Arg242His). GFP-tagged WTSDHBand mutantSDHBconstructs were transfected (HEK293) to determine biological outcomes of these mutantsin vitro. According toin silicopredictions, specificSDHBmutations resulted in impaired mitochondrial localisation and/or SDH enzymatic activity. These results indicated strong genotype–functional correlation forSDHBvariants. This study reveals new insights into the effects ofSDHBmutations and the power of structural modelling in predicting biological consequences. We predict that our functional assessment ofSDHBmutations will serve to better define specific consequences for SDH activity as well as to provide a much needed assay to distinguish pathogenic mutations from benign variants.

IN VITRO/IN SILICO CHARACTERIZATION OF ACTIVE AND PASSIVE STRESSES IN CARDIAC MUSCLE

2012 ◽  
Vol 10 (2) ◽  
pp. 171-188 ◽  
Author(s):  
Markus Boel ◽  
Oscar J. Abilez ◽  
Ahmed N Assar ◽  
Christopher K. Zarins ◽  
Ellen Kuhl
Keyword(s):  
Cardiac Muscle ◽  
In Silico ◽  
In Silico Characterization

Role of 4-O Galloylchlorogenic Acid in Lung Cancer- An Insilico Approach

2017 ◽  
Vol 9 (5) ◽  
Author(s):  
Jaynthy C. ◽  
N. Premjanu ◽  
Abhinav Srivastava
Keyword(s):  
Lung Cancer ◽  
Cell Proliferation ◽  
In Silico ◽  
Computational Study ◽  
Regulation Mechanism ◽  
Down Regulation ◽  
Fruit Extract ◽  
In Vitro Techniques ◽  
Discovery Studio

Cancer is a major disease with millions of patients diagnosed each year with high mortality around the world. Various studies are still going on to study the further mechanisms and pathways of the cancer cell proliferation. Fucosylation is one of the most important oligosaccharide modifications involved in cancer and inflammation. In cancer development increased core fucosylation by FUT8 play an important role in cell proliferation. Down regulation of FUT8 expression may help cure lung cancer. Therefore the computational study based on the down regulation mechanism of FUT8 was mechanised. Sapota fruit extract, containing 4-Ogalloylchlorogenic acid was used as the inhibitor against FUT-8 as target and docking was performed using in-silico tool, Accelrys Discovery Studio. There were several conformations of the docked result, and conformation 1 showed 80% dock score between the ligand and the target. Further the amino acids of the inhibitor involved in docking were studied using another tool, Ligplot. Thus, in-silico analysis based on drug designing parameters shows that the fruit extract can be studied further using in-vitro techniques to know its pharmacokinetics.

Discovery of New AKT1 Inhibitors by Combination of In silico Structure Based Virtual Screening Approaches and Biological Evaluations

2019 ◽  
Author(s):  
Filip Fratev ◽  
Denisse A. Gutierrez ◽  
Renato J. Aguilera ◽  
suman sirimulla
Keyword(s):  
Virtual Screening ◽  
Success Rate ◽  
Protein Interactions ◽  
In Silico ◽  
Biological Data ◽  
In Vitro Binding ◽  
Vitro Binding ◽  
Screening Protocol ◽  
Screening Approaches

AKT1 is emerging as a useful target for treating cancer. Herein, we discovered a new set of ligands that inhibit the AKT1, as shown by in vitro binding and cell line studies, using a newly designed virtual screening protocol that combines structure-based pharmacophore and docking screens. Taking together with the biological data, the combination of structure based pharamcophore and docking methods demonstrated reasonable success rate in identifying new inhibitors (60-70%) proving the success of aforementioned approach. A detail analysis of the ligand-protein interactions was performed explaining observed activities.<br>

Sign in / Sign up

Export Citation Format

Share Document