Microbead analysis of cell binding to immobilized lectin. Part II: Quantitative kinetic profile assay for possible identification of anti-infectivity and anti-cancer reagents

Ghazarian, Anasheh; Oppenheimer, Steven B.

doi:10.1016/j.acthis.2014.07.015

Cited by 4 publications

(4 citation statements)

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“…This could confirm the potential biological activity, as deeper embedding is usually associated with better activity. 48 A previous report showed that melezitose could be used for the development of anticancer therapeutics. 31 Ghazarian and his collaborators performed quantitative kinetic profile assays and revealed that D-melezitose is the best inhibitor of yeast binding to lectin microbeads used as a model system to identify agents that might block cancer cell clumping.…”

Section: -Furanmethanol Is Previously Reported In Malaysianmentioning

confidence: 99%

Acacia Honey-derived Bioactive Compounds Exhibit Induction of p53-dependent Apoptosis in the MCF-7 Human Breast Cancer Cell Line

Hamadou

Bouali

Alhejaili

et al. 2023

Pharmacognosy Magazine

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Background Research studies have focused on discovering new anti-proliferative and pro-apoptotic agents derived from natural products from which honey constitutes a prominent candidate. The Acacia honey (AH) is known to display anticancer activity, but the mechanisms of action are still not well defined. Objectives Using in vitro and computational approaches, we aimed to assess the interaction among selected bioactive compounds derived from AH, with the apoptotic protein p53, which could trigger apoptosis. Methods The phytocompounds of AH were investigated via gas chromatography–mass spectrophotometry analysis. The cytotoxic effect and induced apoptosis on the MCF-7 breast cancer cell line were assessed by 3-(4,5-dimethylthiazolyl-2)-2,5 diphenyltetrazolium bromide and acridine orange-ethidium bromide staining approaches. The molecular docking analysis between AH compounds and p53 was carried out. Results The drug-likeness prediction revealed that most of the identified compounds meet Lipinski’s rules. We demonstrate that AH exerts an interesting cytotoxic effect in a dose-dependent manner against the MCF-7 cell line with IC50 5.053µg/mL. Significant cell alterations and notable induced apoptosis were detected when cells were treated with AH. The molecular docking analysis revealed that melezitose is among the most important potential bioactive compounds that interact with p53 leading to apoptosis. The binding affinity was −8.1 kcal/mol, and the closest molecular interactions in the active site included 10 residues, which could explain the potential biological activity. Conclusion This work sheds light on AH as a significant source of bioactive chemicals with potential for promoting apoptosis that may be exploited as an alternative therapy for breast cancer.

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Section: -Furanmethanol Is Previously Reported In Malaysianmentioning

confidence: 99%

Acacia Honey-derived Bioactive Compounds Exhibit Induction of p53-dependent Apoptosis in the MCF-7 Human Breast Cancer Cell Line

Hamadou

Bouali

Alhejaili

et al. 2023

Pharmacognosy Magazine

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“…They also function in the attachment of pathogens to cells and can cause cell clumping, such as in biofilms and cancer cells that helps cancer cells escape the body's defenses [7][8][9][10]. We use yeast (Saccharomyces cerevisiae) and lectin-derivatized agarose beads to identify glycans that can dissociate yeast from the beads as a model system for identifying glycans that can prevent binding of pathogens to cells [5]. This is done quantitatively by counting the numbers of yeast that remain bound to the lectin beads over time in hundreds of individual experiments in the presence and absence of specific glycans [5].…”

Section: The Yeast Modelmentioning

confidence: 99%

“…Because of its transparency and simplicity, the sea urchin is well suited for research in glycobiology. We have developed new assays using the sea urchin embryo to learn about the roles of specific glycans in mediating cellular interactions [2][3][4][5]. The first assay involves culturing sea urchin embryos in 96 well microplates with and without glycans or glycosidases over time and counting the different morphologies of the developing gut (archenteron) in tens of thousands of living embryos [2,4].…”

Section: The Sea Urchin Nih Designated Model System Microplate Assaymentioning

confidence: 99%

“…We use yeast (Saccharomyces cerevisiae) and lectin-derivatized agarose beads to identify glycans that can dissociate yeast from the beads as a model system for identifying glycans that can prevent binding of pathogens to cells [5]. This is done quantitatively by counting the numbers of yeast that remain bound to the lectin beads over time in hundreds of individual experiments in the presence and absence of specific glycans [5]. In this way glycans are identified that may be clinically useful in anti-adhesion venues.…”

Section: The Yeast Modelmentioning

confidence: 99%

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Simple Novel Assays in Glycobiology

Oppenheimer

2014

J Glycobiol

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In response to an invitation from this journal, I am providing this mini-review of recent work from the Oppenheimer lab. Over the past 4 decades we have developed many assays that help us examine the role of specifc glycans in cellular interactions. Recently we have used two model systems for this work, the sea urchin embryo and yeast (Saccharomyces cerevisiae). We have developed two assays using the sea urchin embryo. One involves a microplate method where living sea urchin embryos are incubated with specific glycans or glycosidases. A specific set of cellular interactions, development of the primitive gut…archenteron, is examined over time in the presence and absence of the sugars or enzymes in living embryos. L-rhamnose and polyglucans have been identified as playing a role in mediating these cellular interactions. The second assay involves microdissection of the primitive gut away from the blastocoel roof to which it adheres.Using independently characterized glycosidases, we showed that polyglucans appear to mediate this cellular interaction. In the second system using yeast, we examine yeast disaggregation from lectin-derivatized agarose beads in the presence and absence of specific glycans using a quantitative, kinetic graphic profile assay.We found that D-melezitose was the best adhesion inhibitor and may be therapeutically useful in anti-adhesion venues of pathogen binding to cells and in cancer cell binding. The Sea Urchin, NIH Designated Model System Microplate assayWe have used the sea urchin embryo in our glycobiology research for over 4 decades. The sea urchin has been designated by NIH as a model system for understanding mechanisms of importance in human health and disease [1]. Because of its transparency and simplicity, the sea urchin is well suited for research in glycobiology. We have developed new assays using the sea urchin embryo to learn about the roles of specific glycans in mediating cellular interactions [2][3][4][5]. The first assay involves culturing sea urchin embryos in 96 well microplates with and without glycans or glycosidases over time and counting the different morphologies of the developing gut (archenteron) in tens of thousands of living embryos [2,4]. The developing gut is a simple model for investigating cellular interactions. Because molecules easily enter the interior of sea urchin embryos [6], we are able to precisely quantify the effects of the added reagents on specific cellular interactions. Using these methods we have developed statistically evaluated, quantitative, kinetic graphic profiles of the effects of glycans and glycosidases on the development of the sea urchin archenteron, a model set of cellular interactions, and found that L-rhamnose and polyglucans appear to be involved in these cellular interactions. Of importance is that we independently characterize enzymes used in our studies and test for unit activity and contaminants.Assay scheme: Grow sea urchin embryos to 24 hrs or 32 hrs post fertilization in normal artificial sea water. In 96 well micropl...

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The forgotten sugar: A review on multifarious applications of melezitose

Balaji

2021

Carbohydrate Research

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Microbead analysis of cell binding to immobilized lectin. Part II: Quantitative kinetic profile assay for possible identification of anti-infectivity and anti-cancer reagents

Cited by 4 publications

References 7 publications

Acacia Honey-derived Bioactive Compounds Exhibit Induction of p53-dependent Apoptosis in the MCF-7 Human Breast Cancer Cell Line

Acacia Honey-derived Bioactive Compounds Exhibit Induction of p53-dependent Apoptosis in the MCF-7 Human Breast Cancer Cell Line

Simple Novel Assays in Glycobiology

The forgotten sugar: A review on multifarious applications of melezitose

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