scholarly journals Investigation of a Potential Scintigraphic Tracer for Imaging Apoptosis: Radioiodinated Annexin V-Kunitz Protease Inhibitor Fusion Protein

2011 ◽  
Vol 2011 ◽  
pp. 1-8 ◽  
Author(s):  
Mei-Hsiu Liao ◽  
Tong-Rong Jan ◽  
Chao-Chih Chiang ◽  
Kuo-Chen Yen ◽  
Tse-Zung Liao ◽  
...  
Keyword(s):  
Fusion Protein ◽  
Protease Inhibitor ◽  
Annexin V ◽  
High Yield ◽  
Apoptotic Cells ◽  
Erythrocyte Ghosts ◽  
Kunitz Protease Inhibitor ◽  
Apoptosis Detection ◽  
Iodine 123 ◽  
Iodine 125

Radiolabeled annexin V (ANV) has been widely used for imaging cell apoptosis. Recently, a novel ANV-Kunitz-type protease inhibitor fusion protein, ANV-6L15, was found to be a promising probe for improved apoptosis detection based on its higher affinity to phosphatidylserine (PS) compared to native ANV. The present paper investigates the feasibility of apoptosis detection using radioiodinated ANV-6L15. Native ANV and ANV-6L15 were labeled with iodine-123 and iodine-125 using Iodogen method. The binding between the radioiodinated proteins and erythrocyte ghosts or chemical-induced apoptotic cells was examined. ANV-6L15 can be radioiodinated with high yield (40%−60%) and excellent radiochemical purity (>95%).123I-ANV-6L15 exhibited a higher binding ratio to erythrocyte ghosts and apoptotic cells compared to123I-ANV. The biodistribution of123I-ANV-6L15 in mice was also characterized.123I-ANV-6L15 was rapidly cleared from the blood. High uptake in the liver and the kidneys may limit the evaluation of apoptosis in abdominal regions. Our data suggest that radiolabled ANV-6L15 may be a better scintigraphic tracer than native ANV for apoptosis detection.

P4-153: The disruption of A-Beta peptide by transthyretin: A mechanism sensitive to Kunitz Protease Inhibitor

2009 ◽  
Vol 5 (4S_Part_16) ◽  
pp. P476-P476
Author(s):  
Rita Costa ◽  
Frederico Ferreira-da Silva ◽  
Maria Joao Saraiva ◽  
Isabel Cardoso
Keyword(s):  
Protease Inhibitor ◽  
Kunitz Protease Inhibitor ◽  
Beta Peptide

A Bacillus subtilis fusion protein system to produce soybean Bowman–Birk protease inhibitor

2007 ◽  
Vol 55 (1) ◽  
pp. 40-52 ◽  
Author(s):  
Gudrun Vogtentanz ◽  
Katherine D. Collier ◽  
Michael Bodo ◽  
Judy H. Chang ◽  
Anthony G. Day ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Fusion Protein ◽  
Protease Inhibitor ◽  
Protein System

High Yield of Active STb Enterotoxin from a Fusion Protein (MBP-STb) Expressed in Escherichia coli

1993 ◽  
Vol 4 (4) ◽  
pp. 275-281 ◽  
Author(s):  
C.E. Handl ◽  
J. Harel ◽  
J.I. Flock ◽  
J.D. Dubreuil
Keyword(s):  
Escherichia Coli ◽  
Fusion Protein ◽  
High Yield

scholarly journals Total Phenols, Identification of Active Compounds and Anticancer Activity of Salvia judaica Boiss against the breast Cancer Cell MDA-231

2021 ◽  
Keyword(s):  
Breast Cancer ◽  
Cell Cycle ◽  
Essential Oil ◽  
Anticancer Activity ◽  
Breast Cancer Cell ◽  
Annexin V ◽  
Ethanol Extract ◽  
G1 Phase ◽  
Apoptotic Cells ◽  
Total Phenols

Salvia judaica is an annual herb from genus Salvia L.; the largest genera of Lamiaceae. It’s a medicinal plant prominent in pharmaceutical applications in many countries around the world. This study aimed to explore bioactive compounds likely to be responsible for the plant anticancer activity, and evaluate anticancer effects, after determining the total content of phenols in the ethanol extract and essential oil in this species. Ethanol extract (EE) and essential oil (EO) were prepared from dried aerial parts (leaves and the flower). GC-MS analysis of EO showed the presence of/43/ effective compounds in varying proportions, the major compounds were sesquiterpenes like delta-cadinene, alpha-Gurjunene, beta-humulene, and alpha-caryophyllene. This is the first study revealed that S.judaica is so rich in phenols which proceeded S.officinalis, noting the superiority of the EE over the EO samples in the total phenols. Anticancer properties of EE and EO of S. judaica against MDA-231 breast cancer cell line were studied -for the first time - by cell cycle analysis and Annexin V/PI apoptosis assay using Flow cytometry technique. Cells were treated with EE (0.001, 0.01, 0.02, 0.1mg/ml) and EO (0.005, 0.01, 0.02, 0.03, 0.04 mg/ml) at various concentrations for48 h. The results revealed that both EE and EO induced cell cycle arrest at G1-phase. Cells treated with EE and EO for 48h showed increasing the percentage of cells in G1-phase and decreasing the percentage of cells in S-phase with increasing concentration compared with untreated cells (control). Annexin V-FITC/PI assay confirmed that EO and EE were able to induce apoptosis. Cells treated with EOat (0.04 mg/ml) for 48h resulted in apoptotic cells at 96.68%, and necrotic cells at 0.12%, compared with untreated cells. On the other hand, Cells treated with EE at (0.1 mg/ml) for 48h resulted in apoptotic cells at 94.43%, and necrotic cells at 0.47%, compared with control. Results revealed that EO is better than EE as anticancer; treatment with EO resulted in more apoptotic cells and less necrotic cells, and there were significant differences between them. This confirmed that EO contains specific anticancer compounds as showed by GC-MS analysis. However, more studies should be performed to explore antioxidants present in S.judaica and determine the underlying mechanism of their anti-breast cancer properties.

Culture and Maintenance of Human Ovarian Carcinoma Cells for Scrutinizing Anti-cancerous Activities of Various Compounds via Some Potent Molecular Markers

2020 ◽  
pp. 120-125
Keyword(s):  
Molecular Markers ◽  
Ovarian Carcinoma ◽  
Genital Tract ◽  
Female Genital Tract ◽  
Annexin V ◽  
Anticancer Efficacy ◽  
Carcinoma Cell Lines ◽  
Cell Transplants ◽  
Apoptosis Detection ◽  
Human Ovarian Carcinoma

Ovarian carcinoma is the 5th most common type of cancer of gynecologic origin and accounts for about one-fourth of the total malignancies of the female genital tract. Ovarian carcinoma accounts for highest mortality in females due to the development of chemo-resistance against drugs and lack of symptoms and undetectable biomarkers in the early stages of diagnosis. Tumour debulking, chemotherapies, radiotherapies, targeted therapies, immunotherapies and stem cell transplants are some of the measures that have been adopted by the experts for curing the disease but still, full control over the problem has not been achieved. Research on various herbal and chemosynthetic nano-compounds have shown a new light in this regard, as the studies on them so far have revealed that they have anti-proliferative and apoptotic properties that will help in finding new ways to develop drugs for cancer patients. This chapter deals how to culture and maintain the human ovarian carcinoma cell lines in the laboratory which are being procured from cell repositories and then to study the anticancer efficacy of various promising compounds by potent molecular markers like cellcycle progression and annexin V- FITC apoptosis detection.

scholarly journals Comparison of Apoptosis Detection Markers Combined with Macrophage Immunostaining to Study Phagocytosis of Apoptotic Cells in Situ

2006 ◽  
Vol 1 ◽  
pp. 117727190600100 ◽  
Author(s):  
Dorien M. Schrijvers ◽  
Guido R.Y. De Meyer ◽  
Mark M. Kockx ◽  
Arnold G. Herman ◽  
Wim Martinet
Keyword(s):  
Lupus Erythematosus ◽  
Caspase 3 ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Apoptotic Cells ◽  
Immunological Responses ◽  
Apoptosis Detection ◽  
Apoptosis Markers ◽  
Suitable Marker ◽  
Parp 1

Efficient phagocytosis of cells undergoing apoptosis by macrophages is important to prevent immunological responses and development of chronic inflammatory disorders such as systemic lupus erythematosus, cystic fibrosis and atherosclerosis. To study phagocytosis of apoptotic cells (AC) by macrophages in tissue, we validated different apoptosis markers (DNA fragmentation, caspase-3 activation and cleavage of its substrate poly(ADP-ribose)polymerase-1) in combination with macrophage immunostaining. Human tonsils were used as a model because they show a high apoptosis frequency under physiological conditions as well as efficient phagocytosis of AC by macrophages. On the other hand, advanced human atherosclerotic plaques were examined since plaques show severely impaired phagocytosis of AC. Our results demonstrate that the presence of non-phagocytized terminal deoxynucleotidyl transferase end labelling (TUNEL)-positive AC represents a suitable marker of poor phagocytosis by macrophages in situ. Other markers for apoptosis, such as cleavage of caspase-3 or PARP-1, should not be used to assess phagocytosis efficiency, because activation of the caspase cascade and cleavage of their substrates can occur in AC when they have not yet been phagocytized by macrophages.

scholarly journals Annexin V for flow cytometric detection of phosphatidylserine expression on B cells undergoing apoptosis

Blood ◽  
1994 ◽  
Vol 84 (5) ◽  
pp. 1415-1420 ◽  
Author(s):  
G Koopman ◽  
CP Reutelingsperger ◽  
GA Kuijten ◽  
RM Keehnen ◽  
ST Pals ◽  
...  
Keyword(s):  
B Cells ◽  
Chromatin Condensation ◽  
Membrane Damage ◽  
Annexin V ◽  
Fluorescein Isothiocyanate ◽  
General Mechanism ◽  
Apoptotic Cells ◽  
Flow Cytometric ◽  
Burkitt Lymphoma Cell ◽  
Cell Expression

Abstract Apoptosis, or programmed cell death, is a general mechanism for removal of unwanted cells from the immune system. It is characterized by chromatin condensation, a reduction in cell volume, and endonuclease cleavage of DNA into oligonucleosomal length fragments. Apoptosis is also accompanied by a loss of membrane phospholipid asymmetry, resulting in the exposure of phosphatidylserine at the surface of the cell. Expression of phosphatidylserine at the cell surface plays an important role in the recognition and removal of apoptotic cells by macrophages. Here we describe a new method for the detection of apoptotic cells by flow cytometry, using the binding of fluorescein isothiocyanate-labeled annexin V to phosphatidylserine. When Burkitt lymphoma cell lines and freshly isolated germinal center B cells are cultured under apoptosis inducing conditions, all cells showing chromatin condensation strongly stain with annexin V, whereas normal cells are annexin V negative. Moreover, DNA fragmentation is only found in the annexin V-positive cells. The nonvital dye ethidium bromide was found to stain a subpopulation of the annexin V-positive apoptotic cells, increasing with time. Our results indicate that the phase in apoptosis that is characterized by chromatin condensation coincides with phosphatidylserine exposure. Importantly, it precedes membrane damage that might lead to release from the cells of enzymes that are harmful to the surrounding tissues. Annexin V may prove important in further unravelling the regulation of apoptosis.

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