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Article ; Online: Carbonic Anhydrase II Activators in Osteopetrosis Treatment: A Review.

Alkhayal, Zikra / Shinwari, Zakia / Gaafar, Ameera / Alaiya, Ayodele

2023 Volume 45, Issue 2, Page(s) 1373–1386

Abstract: Osteopetrosis is a rare hereditary illness generated by failure in osteoclasts resulting in elevated bone densities. Patients with osteopetrosis possess several complications, like dental caries, earlier teeth loss, delayed eruption, malformed crowns and ...

Abstract	Osteopetrosis is a rare hereditary illness generated by failure in osteoclasts resulting in elevated bone densities. Patients with osteopetrosis possess several complications, like dental caries, earlier teeth loss, delayed eruption, malformed crowns and roots, and lamina dura thickening. Since deficiency of carbonic anhydrase II is a major cause behind osteopetrosis, carbonic anhydrase II activators have a large number of applications in osteopetrosis treatment. There is a lack of a comprehensive review on osteopetrosis, pathogenesis of dental abnormalities, and the role of carbonic anhydrase II activators in osteopetrosis treatment. To address this research gap, the authros perfomed a comprehensive review on osteopetrosis and its types, pathogenesis of dental abnormalities, and the role of carbonic anhydrase II activators in osteopetrosis treatment. A brief introduction to the pathogenesis of dental abnormalities and regeneration is provided in this survey. A discussion of types of osteopetrosis depending on genetic inheritance, such as autosomal dominant, autosomal recessive, and X-linked inheritance osteopetrosis, is presented in this survey. The paper also focuses on the importance of carbonic anhydrase II activators as a potential drug therapy for dental osteopetrosis. In addition, a brief note on the role of azole and fluconazole in treating osteopetrosis is given. Finally, future directions involving gene therapy for dental osteopetrosis are described.
Language	English
Publishing date	2023-02-06
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	2000024-8
ISSN	1467-3045 ; 1467-3037
ISSN (online)	1467-3045
ISSN	1467-3037
DOI	10.3390/cimb45020089
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Article ; Online: Fluconazole-Induced Protein Changes in Osteogenic and Immune Metabolic Pathways of Dental Pulp Mesenchymal Stem Cells of Osteopetrosis Patients.

Alkhayal, Zikra / Shinwari, Zakia / Gaafar, Ameera / Alaiya, Ayodele

International journal of molecular sciences

2023 Volume 24, Issue 18

Abstract: Osteopetrosis is a rare inherited disease caused by osteoclast failure, resulting in increasing bone density in humans. Patients with osteopetrosis possess several dental and cranial complications. Since carbonic anhydrase II (CA-II) deficiency is a ... ...

Abstract	Osteopetrosis is a rare inherited disease caused by osteoclast failure, resulting in increasing bone density in humans. Patients with osteopetrosis possess several dental and cranial complications. Since carbonic anhydrase II (CA-II) deficiency is a major cause of osteopetrosis, CA-II activators might be an attractive potential treatment option for osteopetrosis patients. We conducted comprehensive label-free quantitative proteomics analysis on Fluconazole-treated Dental Pulp Mesenchymal Stem/Stromal Cells from CA-II-Deficient Osteopetrosis Patients. We identified 251 distinct differentially expressed proteins between healthy subjects, as well as untreated and azole-treated derived cells from osteopetrosis patients. Twenty-six (26) of these proteins were closely associated with osteogenesis and osteopetrosis disease. Among them are ATP1A2, CPOX, Ap2 alpha, RAP1B and some members of the RAB protein family. Others include AnnexinA1, 5, PYGL, OSTF1 and PGAM4, all interacting with OSTM1 in the catalytic reactions of HCO3 and the Cl- channel via CAII regulation. In addition, the pro-inflammatory/osteoclast regulatory proteins RACK1, MTSE, STING1, S100A13, ECE1 and TRIM10 are involved. We have identified proteins involved in osteogenic and immune metabolic pathways, including ERK 1/2, phosphatase and ATPase, which opens the door for some CA activators to be used as an alternative drug therapy for osteopetrosis patients. These findings propose that fluconazole might be a potential treatment agent for CAII- deficient OP patients. Altogether, our findings provide a basis for further work to elucidate the clinical utility of azole, a CA activator, as a therapeutic for OP.
MeSH term(s)	Humans ; Fluconazole/pharmacology ; Fluconazole/therapeutic use ; Osteogenesis ; Dental Pulp ; Osteopetrosis/drug therapy ; Azoles ; Metabolic Networks and Pathways ; Mesenchymal Stem Cells ; rap GTP-Binding Proteins
Chemical Substances	Fluconazole (8VZV102JFY) ; Azoles ; RAP1B protein, human (EC 3.6.1.-) ; rap GTP-Binding Proteins (EC 3.6.5.2)
Language	English
Publishing date	2023-09-08
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2019364-6
ISSN	1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online)	1422-0067
ISSN	1422-0067 ; 1661-6596
DOI	10.3390/ijms241813841
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Article ; Online: Sex-Biased Expression of Genes Allocated in the Autosomal Chromosomes: Blood LC-MS/MS Protein Profiling in Healthy Subjects.

Giha, Hayder A / Abdulwahab, Rabab A / Abbas, Jaafar / Shinwari, Zakia / Alaiya, Ayodele

Genetics research

2023 Volume 2023, Page(s) 8822205

Abstract: Background: Sex and gender have a large impact in human health and disease prediction. According to genomic/genetics, men differ from women by a limited number of genes in Y chromosome, while the phenotypes of the 2 sexes differ markedly.: Methods: ... ...

Abstract	Background: Sex and gender have a large impact in human health and disease prediction. According to genomic/genetics, men differ from women by a limited number of genes in Y chromosome, while the phenotypes of the 2 sexes differ markedly. Methods: In this study, serum samples from six healthy Bahraini men and women were analyzed by liquid chromatography-mass spectrometry (LC-MS/MS). Bioinformatics databases and tools were used for protein/peptide (PPs) identification and gene localization. The PPs that differed significantly ( Results: Revealed 20 PPs, 11 were upregulated in women with very high FC (up to 8 folds), and 9 were upregulated in men but with much lower FC. The PPs are encoded by genes located in autosomal chromosomes, indicative of sex-biased gene expression. The only PP related to sex, the sex hormone-binding globulin, was upregulated in women. The remaining PPs were involved in immunity, lipid metabolism, gene expression, connective tissue, and others, with some overlap in function. Conclusions: The upregulated PPs in men or women are mostly reflecting the functon or risk/protection provided by the PPs to the specific sex, e.g., Apo-B100 of LDLC. Finally, the basis of sex-biased gene expression and sex phenotypic differences needs further investigation.
MeSH term(s)	Male ; Humans ; Female ; Chromatography, Liquid ; Healthy Volunteers ; Tandem Mass Spectrometry ; Y Chromosome ; Genome
Language	English
Publishing date	2023-03-11
Publishing country	England
Document type	Journal Article
ZDB-ID	2412681-0
ISSN	1469-5073 ; 0016-6723
ISSN (online)	1469-5073
ISSN	0016-6723
DOI	10.1155/2023/8822205
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Article ; Online: Proteomic Profiling of the First Human Dental Pulp Mesenchymal Stem/Stromal Cells from Carbonic Anhydrase II Deficiency Osteopetrosis Patients

Zikra Alkhayal / Zakia Shinwari / Ameera Gaafar / Ayodele Alaiya

International Journal of Molecular Sciences, Vol 22, Iss 380, p

2021 Volume 380

Abstract: Osteopetrosis is a hereditary disorder characterized by sclerotic, thick, weak, and brittle bone. The biological behavior of mesenchymal cells obtained from osteopetrosis patients has not been well-studied. Isolated mesenchymal stem/stromal cells from ... ...

Abstract	Osteopetrosis is a hereditary disorder characterized by sclerotic, thick, weak, and brittle bone. The biological behavior of mesenchymal cells obtained from osteopetrosis patients has not been well-studied. Isolated mesenchymal stem/stromal cells from dental pulp (DP-MSSCs) of recently extracted deciduous teeth from osteopetrosis (OP) patients and healthy controls (HCs) were compared. We evaluated whether the dental pulp of OP patients has a population of MSSCs with similar multilineage differentiation capability to DP-MSSCs of healthy subjects. Stem/progenitor cells were characterized using immunohistochemistry, flow cytometry, and proteomics. Our DP-MSSCs were strongly positive for CD44, CD73, CD105, and CD90. DP-MSSCs obtained from HC subjects and OP patients showed similar patterns of proliferation and differentiation as well as gene expression. Proteomic analysis identified 1499 unique proteins with 94.3% similarity in global protein fingerprints of HCs and OP patients. Interestingly, we observed subtle differences in expressed proteins of osteopetrosis disease-related in pathways, including MAPK, ERK 1/2, PI3K, and integrin, rather than in the stem cell signaling network. Our findings of similar protein expression signatures in DP-MSSCs of HC and OP patients are of paramount interest, and further in vivo validation study is needed. There is the possibility that OP patients could have their exfoliating deciduous teeth banked for future use in regenerative dentistry.
Keywords	human-exfoliated-deciduous-teeth ; mesenchymal stem/stromal cells (MSSCs) ; proteomics ; osteopetrosis ; carbonic anhydrase II ; dental pulp ; Biology (General) ; QH301-705.5 ; Chemistry ; QD1-999
Subject code	610
Language	English
Publishing date	2021-12-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
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Article ; Online: Proteomic Profiling of the First Human Dental Pulp Mesenchymal Stem/Stromal Cells from Carbonic Anhydrase II Deficiency Osteopetrosis Patients.

Alkhayal, Zikra / Shinwari, Zakia / Gaafar, Ameera / Alaiya, Ayodele

International journal of molecular sciences

2020 Volume 22, Issue 1

Abstract	Osteopetrosis is a hereditary disorder characterized by sclerotic, thick, weak, and brittle bone. The biological behavior of mesenchymal cells obtained from osteopetrosis patients has not been well-studied. Isolated mesenchymal stem/stromal cells from dental pulp (DP-MSSCs) of recently extracted deciduous teeth from osteopetrosis (OP) patients and healthy controls (HCs) were compared. We evaluated whether the dental pulp of OP patients has a population of MSSCs with similar multilineage differentiation capability to DP-MSSCs of healthy subjects. Stem/progenitor cells were characterized using immunohistochemistry, flow cytometry, and proteomics. Our DP-MSSCs were strongly positive for CD44, CD73, CD105, and CD90. DP-MSSCs obtained from HC subjects and OP patients showed similar patterns of proliferation and differentiation as well as gene expression. Proteomic analysis identified 1499 unique proteins with 94.3% similarity in global protein fingerprints of HCs and OP patients. Interestingly, we observed subtle differences in expressed proteins of osteopetrosis disease-related in pathways, including MAPK, ERK 1/2, PI3K, and integrin, rather than in the stem cell signaling network. Our findings of similar protein expression signatures in DP-MSSCs of HC and OP patients are of paramount interest, and further in vivo validation study is needed. There is the possibility that OP patients could have their exfoliating deciduous teeth banked for future use in regenerative dentistry.
MeSH term(s)	Acidosis, Renal Tubular/metabolism ; Acidosis, Renal Tubular/pathology ; Adolescent ; Biomarkers/analysis ; Biomarkers/metabolism ; Carbonic Anhydrases/deficiency ; Carbonic Anhydrases/metabolism ; Case-Control Studies ; Cell Differentiation ; Cell Proliferation ; Child ; Dental Pulp/cytology ; Dental Pulp/metabolism ; Female ; Humans ; Male ; Mesenchymal Stem Cells/cytology ; Mesenchymal Stem Cells/metabolism ; Osteopetrosis/metabolism ; Osteopetrosis/pathology ; Proteome/analysis ; Urea Cycle Disorders, Inborn/metabolism ; Urea Cycle Disorders, Inborn/pathology
Chemical Substances	Biomarkers ; Proteome ; Carbonic Anhydrases (EC 4.2.1.1)
Language	English
Publishing date	2020-12-31
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2019364-6
ISSN	1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online)	1422-0067
ISSN	1422-0067 ; 1661-6596
DOI	10.3390/ijms22010380
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Article ; Online: Poly-guanidine shows high cytotoxicity in glioma cell cultures and glioma stem cells.

Márquez, Marcela / Olausson, Karl Holmberg / Alaiya, Ayodele / Nilsson, Sten / Meurling, Lennart / Holmberg, Anders R

Investigational new drugs

2022 Volume 40, Issue 3, Page(s) 565–575

Abstract: Glioblastoma multiforme (GBM) is a malignant CNS tumor with a poor prognosis. GBM shows aberrant glycosylation with hypersialylation. This property is a potential target for therapy. This study investigates the growth inhibitory efficacy of poly- ... ...

Abstract	Glioblastoma multiforme (GBM) is a malignant CNS tumor with a poor prognosis. GBM shows aberrant glycosylation with hypersialylation. This property is a potential target for therapy. This study investigates the growth inhibitory efficacy of poly-guanidine (GuaDex), with an affinity for sialic acid (Sia). Glioma cell cultures and patient-derived glioma cell lines (PDGCLs) expressing Prominin-1 (CD133) were used. Human fibroblasts and astrocyte-derived cells were used as controls. Temozolomide (standard GBM drug, TMZ) and DMSO were used as a comparison. GuaDex at 1-10 µM concentrations, were incubated for 3.5-72 h and with PDGCLs cells for 6-24 h. The cytotoxicity was estimated with a fluorometric cytotoxicity assay (FMCA). Fluorescence-labelled GuaDex was used to study the cell interactions. Sia expression was confirmed with a fluorescence labelled Sia binding lectin. Expression of glial fibrillary acidic protein was determined. GuaDex induction of growth inhibition was fast, showing after less than 5 min incubation while the control cells were not affected even after 50 min incubation. The growth inhibitory effect on PDGCLs spheroids was persistent still showing after 4 weeks post-treatment. The growth inhibition of GuaDex was induced at low µM concentrations while TMZ induced only a slight inhibition at mM concentrations. GuaDex efficacy appears significant and warrants further studies.
MeSH term(s)	Brain Neoplasms/pathology ; Cell Culture Techniques ; Cell Line, Tumor ; Drug Resistance, Neoplasm ; Glioblastoma/drug therapy ; Glioma/metabolism ; Guanidine/pharmacology ; Guanidine/therapeutic use ; Humans ; Neoplastic Stem Cells ; Temozolomide/pharmacology ; Temozolomide/therapeutic use
Chemical Substances	Guanidine (JU58VJ6Y3B) ; Temozolomide (YF1K15M17Y)
Language	English
Publishing date	2022-03-21
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	604895-x
ISSN	1573-0646 ; 0167-6997
ISSN (online)	1573-0646
ISSN	0167-6997
DOI	10.1007/s10637-022-01233-7
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Article ; Online: Cytotoxicity of poly-guanidine in medulloblastoma cell lines.

Gallo-Oller, Gabriel / de Ståhl, Teresita Díaz / Alaiya, Ayodele / Nilsson, Sten / Holmberg, Anders R / Márquez-Méndez, Marcela

Investigational new drugs

2023 Volume 41, Issue 5, Page(s) 688–698

Abstract: Medulloblastoma (MB) is the most common pediatric brain tumor. The therapy frequently causes serious side effects, and new selective therapies are needed. MB expresses hyper sialylation, a possible target for selective therapy. The cytotoxic efficacy of ... ...

Abstract	Medulloblastoma (MB) is the most common pediatric brain tumor. The therapy frequently causes serious side effects, and new selective therapies are needed. MB expresses hyper sialylation, a possible target for selective therapy. The cytotoxic efficacy of a poly guanidine conjugate (GuaDex) incubated with medulloblastoma cell cultures (DAOY and MB-LU-181) was investigated. The cells were incubated with 0.05-8 µM GuaDex from 15 min to 72 h. A fluorometric cytotoxicity assay (FMCA) measured the cytotoxicity. Labeled GuaDex was used to study tumor cell interaction. FITC-label Sambucus nigra confirmed high expression of sialic acid (Sia). Immunofluorescence microscopy was used to visualize the cell F-actin and microtubules. The cell interactions were studied by confocal and fluorescence microscopy. Annexin-V assay was used to detect apoptosis. Cell cycle analysis was done by DNA content determination. A wound-healing migration assay determined the effects on the migratory ability of DAOY cells after GuaDex treatment. IC
MeSH term(s)	Child ; Humans ; Medulloblastoma/drug therapy ; Medulloblastoma/genetics ; Medulloblastoma/metabolism ; Guanidine/pharmacology ; Guanidine/therapeutic use ; Fluorescein-5-isothiocyanate/pharmacology ; Fluorescein-5-isothiocyanate/therapeutic use ; Cell Proliferation ; Cell Line, Tumor ; Apoptosis ; Cerebellar Neoplasms/drug therapy ; Cerebellar Neoplasms/metabolism ; Cerebellar Neoplasms/pathology ; DNA
Chemical Substances	Guanidine (JU58VJ6Y3B) ; Fluorescein-5-isothiocyanate (I223NX31W9) ; DNA (9007-49-2)
Language	English
Publishing date	2023-08-09
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	604895-x
ISSN	1573-0646 ; 0167-6997
ISSN (online)	1573-0646
ISSN	0167-6997
DOI	10.1007/s10637-023-01386-z
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Article ; Online: Anti-Inflammatory Effect of Specialized Proresolving Lipid Mediators on Mesenchymal Stem Cells: An In Vitro Study.

AlZahrani, Shahd / Shinwari, Zakia / Gaafar, Ameera / Alaiya, Ayodele / Al-Kahtani, Ahmed

Cells

2022 Volume 12, Issue 1

Abstract: An interconnection between tissue inflammation and regeneration has been established through the regulation of defense and repair mechanisms within diseased dental tissue triggered by the release of immune-resolvent mediators. To better our understanding ...

Abstract	An interconnection between tissue inflammation and regeneration has been established through the regulation of defense and repair mechanisms within diseased dental tissue triggered by the release of immune-resolvent mediators. To better our understanding of the role of specific pro-resolving mediators (SPMs) in inflamed human bone marrow-derived mesenchymal stem cells (hBMMSCs), we studied the effects of Resolvin E1 (RvE1) and Maresin 1 (MaR1) in lipopoly-saccharide (LPS) stimulated hBMMSCs. The hBMMSCs were divided into five different groups, each of which was treated with or without SPMs. Group-1: negative control (no LPS stimulation), Group-2: positive control (LPS-stimulated), Group-3: RvE1 100 nM + 1 μg/mL LPS, Group-4: MaR1 100 nM + 1 µg/mL LPS, and Group-5: RvE1 100 nM + MaR1100 nM + 1 μg/mL LPS. Cell proliferation, apoptosis, migration, colony formation, Western blotting, cytokine array, and LC/MS analysis were all performed on each group to determine the impact of SPMs on inflammatory stem cells. According to our data, RvE1 plus MaR1 effectively reduced inflammation in hBMMSCs. In particular, IL-4, 1L-10, and TGF-β1 activation and downregulation of RANKL, TNF-α, and IFN-γ compared to groups receiving single SPM were shown to be significantly different (Group 3 and 4). In addition, the LC/MS analysis revealed the differentially regulated peptide's role in immunological pathways that define the cellular state against inflammation. Inflamed hBMMSCs treated with a combination of Resolvin E1 (RvE1) and Maresin 1 (MaR1) promoted the highest inflammatory resolution compared to the other groups; this finding suggests a potential new approach of treating bacterially induced dental infections.
MeSH term(s)	Humans ; Eicosanoids ; Inflammation/metabolism ; Cytokines ; Anti-Inflammatory Agents/pharmacology ; Anti-Inflammatory Agents/therapeutic use ; Mesenchymal Stem Cells/metabolism
Chemical Substances	Eicosanoids ; Cytokines ; Anti-Inflammatory Agents
Language	English
Publishing date	2022-12-28
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2661518-6
ISSN	2073-4409 ; 2073-4409
ISSN (online)	2073-4409
ISSN	2073-4409
DOI	10.3390/cells12010122
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Article ; Online: Crosstalk between aryl hydrocarbon receptor (AhR) and BCL-2 pathways suggests the use of AhR antagonist to maintain normal differentiation state of mammary epithelial cells during BCL-2 inhibition therapy.

Al-Dhfyan, Abdullah / Alaiya, Ayodele / Al-Mohanna, Falah / Attwa, Mohamed W / AlAsmari, Abdullah F / Bakheet, Saleh A / Korashy, Hesham M

Journal of advanced research

2022 Volume 50, Page(s) 177–192

Abstract: Introduction: Activating the aryl hydrocarbon receptor upon exposure to environmental pollutants promotes development of breast cancer stem cell (CSCs). BCL-2 family proteins protect cancer cells from the apoptotic effects of chemotherapeutic drugs. ... ...

Abstract	Introduction: Activating the aryl hydrocarbon receptor upon exposure to environmental pollutants promotes development of breast cancer stem cell (CSCs). BCL-2 family proteins protect cancer cells from the apoptotic effects of chemotherapeutic drugs. However, the crosstalk between AhR and the BCL-2 family in CSC development remains uninvestigated. Objectives: This study explored the interaction mechanisms between AhR and BCL-2 in CSC development and chemoresistance. Methods: A quantitative proteomic analysis study was performed as a tool for comparative expression analysis of breast cancer cells treated by AhR agonist. The basal and inducible levels of BCL-2, AhR, and CYP1A1 in vitro breast cancer and epithelial cell lines and in vivo mice animal models were determined by RT-PCR, Western blot analysis, immunofluorescence, flow cytometry, silencing of the target, and immunohistochemistry. In addition, an in silico toxicity study was conducted using DEREK software. Results: Activation of the AhR/CYP1A1 pathway in mice increased EpCAM Conclusions: This is the first study to report crosstalk between AhR and BCL-2 in breast CSCs and provides the rationale for using a combined treatment of BCL-2 inhibitor and AhR antagonist for more effective cancer prevention and treatment.
MeSH term(s)	Mice ; Animals ; Receptors, Aryl Hydrocarbon/metabolism ; Epithelial Cell Adhesion Molecule ; Integrin alpha6 ; Cell Line, Tumor ; Cytochrome P-450 CYP1A1/metabolism ; Proteomics ; Epithelial Cells/metabolism ; Cell Differentiation
Chemical Substances	Receptors, Aryl Hydrocarbon ; Epithelial Cell Adhesion Molecule ; Integrin alpha6 ; Cytochrome P-450 CYP1A1 (EC 1.14.14.1)
Language	English
Publishing date	2022-10-26
Publishing country	Egypt
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2541849-X
ISSN	2090-1224 ; 2090-1224
ISSN (online)	2090-1224
ISSN	2090-1224
DOI	10.1016/j.jare.2022.10.006
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Article ; Online: Inhibition of signaling downstream of beta-2 adrenoceptor by propranolol in prostate cancer cells.

Alaskar, Aljoharah / Abdulraqeb Ali, Amaal / Hassan, Sazzad / Shinwari, Zakia / Alaiya, Ayodele / von Holzen, Urs / Miller, Lance / Kulik, George

The Prostate

2022 Volume 83, Issue 3, Page(s) 237–245

Abstract: Background: There is accumulating evidence that propranolol, an antagonist of beta-1 and beta-2 adrenoreceptors, extends survival of patients with prostate cancer; yet it is not known whether propranolol inhibits beta-adrenergic signaling in prostate ... ...

Abstract	Background: There is accumulating evidence that propranolol, an antagonist of beta-1 and beta-2 adrenoreceptors, extends survival of patients with prostate cancer; yet it is not known whether propranolol inhibits beta-adrenergic signaling in prostate cancer cells, or systemic effects of propranolol play the leading role in slowing down cancer progression. Recently initiated clinical studies offer a possibility to test whether administration of propranolol inhibits signaling pathways in prostate tumors, however, there is limited information on the dynamics of signaling pathways activated downstream of beta-2 adrenoreceptors in prostate cancer cells and on the inactivation of these pathways upon propranolol administration. Methods: Western blot analysis was used to test the effects of epinephrine and propranolol on activation of protein kinase (PKA) signaling in mouse prostates and PKA, extracellular signal-regulated kinase (ERK), and protein kinase B/AKT (AKT) signaling in prostate cancer cell lines. Results: In prostate cancer cell lines epinephrine induced robust phosphorylation of PKA substrates pS133CREB and pS157VASP that was evident 2 min after treatments and lasted for 3-6 h. Epinephrine induced phosphorylation of AKT in PTEN-positive 22Rv1 cells, whereas changes of constitutive AKT phosphorylation were minimal in PTEN-negative PC3, C42, and LNCaP cells. A modest short-term increase of pERK in response to epinephrine was observed in all tested cell lines. Incubation of prostate cancer cells with 10-fold molar excess of propranolol for 30 min inhibited all downstream pathways activated by epinephrine. Subjecting mice to immobilization stress induced phosphorylation of S133CREB, whereas injection of propranolol at 1.5 mg/kg prevented the stress-induced phosphorylation. Conclusions: The analysis of pS133CREB and pS157VASP allows measuring activation of PKA signaling downstream of beta-2 adrenoreceptors. Presented results on the ratio of propranolol/epinephrine and the time needed to inhibit signaling downstream of beta-2 adrenoreceptors will help to design clinical studies that examine the effects of propranolol on prostate tumors.
MeSH term(s)	Humans ; Male ; Animals ; Mice ; Propranolol/pharmacology ; Propranolol/metabolism ; Proto-Oncogene Proteins c-akt/metabolism ; Prostate/pathology ; Prostatic Neoplasms/pathology ; Phosphorylation ; Epinephrine/pharmacology ; Epinephrine/metabolism
Chemical Substances	Propranolol (9Y8NXQ24VQ) ; Proto-Oncogene Proteins c-akt (EC 2.7.11.1) ; Epinephrine (YKH834O4BH)
Language	English
Publishing date	2022-11-14
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	604707-5
ISSN	1097-0045 ; 0270-4137
ISSN (online)	1097-0045
ISSN	0270-4137
DOI	10.1002/pros.24455
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