Experimental Approaches to Studying O-Glycosylation In Vivo

1997 ◽  
pp. 313-322
Keyword(s):  
Approaches To Studying ◽  
Experimental Approaches

scholarly journals Discovery of vanoxerine dihydrochloride as a CDK2/4/6 triple-inhibitor for the treatment of human hepatocellular carcinoma

2021 ◽  
Vol 27 (1) ◽  
Author(s):  
Ying Zhu ◽  
Kun-Bin Ke ◽  
Zhong-Kun Xia ◽  
Hong-Jian Li ◽  
Rong Su ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Cell Cycle Progression ◽  
G1 Arrest ◽  
Huh7 Cells ◽  
Synergistic Cytotoxicity ◽  
Combination Study ◽  
Experimental Approaches ◽  
Induced Apoptosis

Abstract Background Cyclin-dependent kinases 2/4/6 (CDK2/4/6) play critical roles in cell cycle progression, and their deregulations are hallmarks of hepatocellular carcinoma (HCC). Methods We used the combination of computational and experimental approaches to discover a CDK2/4/6 triple-inhibitor from FDA approved small-molecule drugs for the treatment of HCC. Results We identified vanoxerine dihydrochloride as a new CDK2/4/6 inhibitor, and a strong cytotoxicdrugin human HCC QGY7703 and Huh7 cells (IC50: 3.79 μM for QGY7703and 4.04 μM for Huh7 cells). In QGY7703 and Huh7 cells, vanoxerine dihydrochloride treatment caused G1-arrest, induced apoptosis, and reduced the expressions of CDK2/4/6, cyclin D/E, retinoblastoma protein (Rb), as well as the phosphorylation of CDK2/4/6 and Rb. Drug combination study indicated that vanoxerine dihydrochloride and 5-Fu produced synergistic cytotoxicity in vitro in Huh7 cells. Finally, in vivo study in BALB/C nude mice subcutaneously xenografted with Huh7 cells, vanoxerine dihydrochloride (40 mg/kg, i.p.) injection for 21 days produced significant anti-tumor activity (p < 0.05), which was comparable to that achieved by 5-Fu (10 mg/kg, i.p.), with the combination treatment resulted in synergistic effect. Immunohistochemistry staining of the tumor tissues also revealed significantly reduced expressions of Rb and CDK2/4/6in vanoxerinedihydrochloride treatment group. Conclusions The present study isthe first report identifying a new CDK2/4/6 triple inhibitor vanoxerine dihydrochloride, and demonstrated that this drug represents a novel therapeutic strategy for HCC treatment.

In vitro assessment of food-derived-glucose bioaccessibility and bioavailability in bicameral cell culture system

2020 ◽  
Vol 45 (5) ◽  
pp. 631-637
Author(s):  
Cansu Ozel-Tasci ◽  
Gozde Pilatin ◽  
Ozgur Edeer ◽  
Sukru Gulec
Keyword(s):  
Cell Culture ◽  
Culture System ◽  
Metabolic Diseases ◽  
Enzymatic Digestion ◽  
Cell Culture System ◽  
Culture Studies ◽  
Experimental Approaches ◽  
In Vitro Assessment

AbstractBackgroundFunctional foods can help prevent metabolic diseases, and it is essential to evaluate functional characteristics of foods through in vitro and in vivo experimental approaches.ObjectiveWe aimed to use the bicameral cell culture system combined with the in vitro digestion to evaluate glucose bioavailability.Materials and methodsCake, almond paste, and pudding were modified by adding fiber and replacing sugar with sweeteners and polyols. Digestion process was modeled in test tubes. Rat enterocyte cells (IEC-6) were grown in a bicameral cell culture system to mimic the physiological characteristics of the human intestine. The glucose bioaccessibility and cellular glucose efflux were measured by glucose oxidase assay.Results and discussionThe glucose bioaccessibilities of modified foods were significantly lower (cake: 2.6 fold, almond paste: 9.2 fold, pudding 2.8 fold) than the controls. Cellular glucose effluxes also decreased in the modified cake, almond paste, and pudding by 2.2, 4, and 2 fold respectively compared to their controls.ConclusionOur results suggest that combining in vitro enzymatic digestion with cell culture studies can be a practical way to test in vitro glucose bioaccessibility and bioavailability in functional food development.

Evolution of moonlighting proteins: insight from yeasts

2014 ◽  
Vol 42 (6) ◽  
pp. 1715-1719 ◽  
Author(s):  
Carlos Gancedo ◽  
Carmen-Lisset Flores ◽  
Juana M. Gancedo
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Present Article ◽  
Catabolite Repression ◽  
Kluyveromyces Lactis ◽  
Approaches To Studying ◽  
Moonlighting Proteins ◽  
Experimental Approaches ◽  
Gal Genes

The present article addresses the possibilities offered by yeasts to study the problem of the evolution of moonlighting proteins. It focuses on data available on hexokinase from Saccharomyces cerevisiae that moonlights in catabolite repression and on galactokinase from Kluyveromyces lactis that moonlights controlling the induction of the GAL genes. Possible experimental approaches to studying the evolution of moonlighting hexose kinases are suggested.

scholarly journals Antitumor Activity and Mechanism of Action of the Hormonotoxin, an LHRH Analog Conjugated to Dermaseptin-B2, a Multifunctional Antimicrobial Peptide

2021 ◽  
Author(s):  
Mickael COUTY ◽  
Marie DUSAUD ◽  
Mickael MIRO-PADOVANI ◽  
Liuhui ZHANG ◽  
Patricia ZADIGUE ◽  
...  
Keyword(s):  
Antitumor Activity ◽  
Antimicrobial Peptide ◽  
Tumor Cell Line ◽  
Experimental Approaches ◽  
Prostate Cancers ◽  
Innovative Drugs ◽  
Therapeutic Alternatives ◽  
Prostate Tumor Cells

Abstract Prostate cancer represents the most common cancer in men. For patients with advanced or metastatic form, treatments will be able to slow down the progression but cannot cure it even with the used of new targeted therapies. In this context, the development of innovative drugs resulting from the exploration of biodiversity could open new therapeutic alternatives. Dermaseptin-B2 (DRS-B2), a natural multifunctional antimicrobial peptide isolated from the Amazonian frog skin, has been reported to possess antitumor and antiangiogenic activities. To improve DRS-B2 pharmacological properties and target prostate tumor cells more specifically, we have developed a chimeric molecule, called Hormonotoxin (H-B2) which is composed of a DRS-B2 combined with a hormonal analog, d-Lys6-LHRH, to target LHRH-Receptor which is overexpressed in more than 85% of prostate cancers. In vitro H-B2 has a significant antiproliferative effect on the PC3 tumor cell line, with an IC50 value close to that of DRS-B2. The antitumor activity of H-B2 was confirmed in vivo in mouse model xenografted with PC3 tumors and appears to be better tolerated than DRS-B2. Biophysical experiments showed that the addition of the hormonal analog to DRS-B2 did not alter either its secondary structure or its biological activity. Combination of different experimental approaches indicated that H-B2 induces cell death by an apoptotic mechanism whereas DRS-B2 was shown to induce it by necrosis. These results could explain the H-B2 less toxicity compared to DRS-B2. H-B2 represents a promising targeting approach for cancer therapy.

scholarly journals SPARC: a method to genetically manipulate precise proportions of cells

2019 ◽  
Author(s):  
Jesse Isaacman-Beck ◽  
Kristine C. Paik ◽  
Carl F. R. Wienecke ◽  
Helen H. Yang ◽  
Yvette E. Fisher ◽  
...  
Keyword(s):  
Gene Expression ◽  
Calcium Imaging ◽  
Transgene Expression ◽  
Cell Populations ◽  
Individual Animal ◽  
Cell Type ◽  
Targeted Expression ◽  
Experimental Approaches ◽  
Mitotic Cells

AbstractMany experimental approaches rely on controlling gene expression in select subsets of cells within an individual animal. However, reproducibly targeting transgene expression to specific fractions of a genetically-defined cell-type is challenging. We developed Sparse Predictive Activity through Recombinase Competition (SPARC), a generalizable toolkit that can express any effector in precise proportions of post-mitotic cells in Drosophila. Using this approach, we demonstrate targeted expression of many effectors and apply these tools to calcium imaging of individual neurons and optogenetic manipulation of sparse cell populations in vivo.

scholarly journals Antitumor Activity and Mechanism of Action of Hormonotoxin, an LHRH Analog Conjugated to Dermaseptin-B2, a Multifunctional Antimicrobial Peptide

2021 ◽  
Vol 22 (21) ◽  
pp. 11303
Author(s):  
Mickael Couty ◽  
Marie Dusaud ◽  
Mickael Miro-Padovani ◽  
Liuhui Zhang ◽  
Patricia Zadigue ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Antitumor Activity ◽  
Antimicrobial Peptide ◽  
Tumor Cell Line ◽  
Ic50 Value ◽  
Experimental Approaches ◽  
Innovative Drugs ◽  
Therapeutic Alternatives ◽  
Apoptotic Mechanism

Prostate cancer is the most common cancer in men. For patients with advanced or metastatic prostate cancer, available treatments can slow down its progression but cannot cure it. The development of innovative drugs resulting from the exploration of biodiversity could open new therapeutic alternatives. Dermaseptin-B2, a natural multifunctional antimicrobial peptide isolated from Amazonian frog skin, has been reported to possess antitumor activity. To improve its pharmacological properties and to decrease its peripheral toxicity and lethality we developed a hormonotoxin molecule composed of dermaseptin-B2 combined with d-Lys6-LHRH to target the LHRH receptor. This hormonotoxin has a significant antiproliferative effect on the PC3 tumor cell line, with an IC50 value close to that of dermaseptin-B2. Its antitumor activity has been confirmed in vivo in a xenograft mouse model with PC3 tumors and appears to be better tolerated than dermaseptin-B2. Biophysical experiments showed that the addition of LHRH to dermaseptin-B2 did not alter its secondary structure or biological activity. The combination of different experimental approaches indicated that this hormonotoxin induces cell death by an apoptotic mechanism instead of necrosis, as observed for dermaseptin-B2. These results could explain the lower toxicity observed for this hormonotoxin compared to dermaseptin-B2 and may represent a promising targeting approach for cancer therapy.

SECRETION OF PROGESTERONE BY HUMAN OVARIES PERFUSED IN VITRO

1975 ◽  
Vol 79 (1) ◽  
pp. 111-121 ◽  
Author(s):  
H. Fukunishi ◽  
H. Mickan ◽  
J. Zander
Keyword(s):  
Ovarian Function ◽  
Sodium Lactate ◽  
Bicarbonate Buffer ◽  
Ldh Activity ◽  
Progesterone Secretion ◽  
Experimental Approaches ◽  
Lactate And Pyruvate ◽  
Human Ovaries

ABSTRACT A system for the perfusion of isolated human ovaries is described. Krebs-Ringer bicarbonate buffer (pH 7.4) containing glucose, sodium lactate and pyruvate was perfused at a flow rate of 5–8 ml/min at 37°C through the ovarian artery. Oxygen uptake, LDH-activity, pH and progesterone secretion were determined as parameters of viability of the perfused organ. Our results demonstrate that a relatively simple system used for perfusion was able to preserve ovarian function for at least two hours. This experimental design should not be regarded as physiological but in addition to other experimental approaches it might provide information on the physiology of the human ovary. Though lower values of progesterone secretion than in vivo were found, a correlation of secretion and functional stage of the ovaries could be demonstrated.

Experimental approaches to studying the photostability of selected pesticides in water and soil

Chemosphere ◽  
1997 ◽  
Vol 35 (1-2) ◽  
pp. 39-50 ◽  
Author(s):  
M. Mansour ◽  
E.A. Feicht ◽  
A. Behechti ◽  
I. Scheunert
Keyword(s):  
Approaches To Studying ◽  
Pesticides In Water ◽  
Experimental Approaches
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