scholarly journals Effects of spermatozoa–oviductal cell coincubation time and oviductal cell age on spermatozoa–oviduct interactions

2014 ◽  
Vol 26 (2) ◽  
pp. 358 ◽  
Author(s):  
Ahmed Aldarmahi ◽  
Sarah Elliott ◽  
Jean Russell ◽  
Alireza Fazeli
Keyword(s):  
Gene Expression ◽  
Quantitative Polymerase Chain Reaction ◽  
Cell Aging ◽  
Control Group ◽  
Prostaglandin E ◽  
Cell Age ◽  
Cell Gene Expression ◽  
Passage Number ◽  
Cell Culture Passage ◽  
Oviductal Cell

The oviduct plays a crucial role in sperm storage, maintenance of sperm viability and sperm transport to the site of fertilisation. The aim of the present study was to investigate the effects of oviductal cell culture passage number, oviductal cell age and spermatozoa–oviduct coincubation times on gene expression in oviductal cells. Immortalised oviductal epithelial cells (OPEC) obtained from two different cell passages (36 and 57) were subcultured three times with and without spermatozoa for 24 h (control group). In a second study, OPEC were cocultured with spermatozoa for different time intervals (0, 4, 12 and 24 h). Expression of adrenomedullin (ADM), heat shock 70 kDa protein 8 (HSPA8) and prostaglandin E synthase (PGES) in OPEC was measured by quantitative polymerase chain reaction. The expression of ADM and HSPA8 was decreased significantly in OPEC cells from Passage 57, particularly in the later subculture group. These effects on HSPA8, but not ADM, expression in OPEC were further altered after coculture with spermatozoa for 24 h. We also demonstrated that spermatozoa–oviduct coculture for 12 and 24 h resulted in significantly higher expression of ADM, HSPA8 and PGES in OPEC. Overall, the data suggest that the OPEC lose some of their properties as a result of oviductal cell aging and that there are spermatozoa–oviduct interactions leading to increased oviductal cell gene expression.

ATF3 and EGR2 gene expression levels in sdLDL-treated macrophages of patients with coronary artery stenosis

2018 ◽  
Vol 42 (1-2) ◽  
pp. 23-29
Author(s):  
Sayed R. Hosseini-Fard ◽  
Mohsen Khosravi ◽  
Amaneh Yarnazari ◽  
Parisa Hassanpour ◽  
Abdollah Amirfarhangi ◽  
...  
Keyword(s):  
Gene Expression ◽  
Coronary Artery ◽  
Cholesterol Metabolism ◽  
Quantitative Polymerase Chain Reaction ◽  
Fold Change ◽  
Expression Level ◽  
Control Group ◽  
Expression Levels ◽  
Vessel Disease ◽  
Gene Expression Levels

AbstractBackground:The metabolism of cholesteryl esters (CEs) is under the control of a gene network in macrophages. Several genes such asATF3andEGR2are related to cholesterol metabolism.Methods:In this study, theATF3andEGR2gene expression levels were evaluated in differentiated macrophages of subjects undergoing coronary artery angiography [controls (<5% stenosis), patients (>70% stenosis)] after treatment with small dense low density lipoprotein (sdLDL) particles. Monocytes were isolated using a RosetteSep Kit and were differentiated into macrophages using the M-CSF factor. A modified heparin-MgSO4-PEG method was used for the sdLDL preparation. TheATF3andEGR2gene expression levels were measured by the real-time quantitative polymerase chain reaction (RT-qPCR) technique.Results:In contrast with the control group (p=0.4), theATF3expression level reduced significantly in the differentiated macrophages from all patients [single vessel disease (SVD), fold change 17 times, p=0.02; two vessel disease (2VD), fold change 1.5 times, p=0.05; three vessel disease (3VD), fold change 3.5 times, p=0.04]. Also, theEGR2expression level reduced significantly in all groups (p<0.02). The gene fold changes had no significant differences between the patients (p>0.8).Conclusions:We propose that the failure ofATF3gene expression improves the CE synthesis after sdLDL influx. Furthermore, the reducedEGR2gene expression level in the sdLDL-treated groups may be a negative factor in cholesterol homeostasis.

scholarly journals Interleukin gene expression in broiler chickens infected by different Escherichia coli serotypes

2021 ◽  
pp. 2727-2734
Author(s):  
Reham Elnagar ◽  
Rasha Elkenany ◽  
Gamal Younis
Keyword(s):  
Gene Expression ◽  
Escherichia Coli ◽  
Broiler Chickens ◽  
Quantitative Polymerase Chain Reaction ◽  
Clinical Signs ◽  
Negative Control Group ◽  
Negative Control ◽  
Control Group ◽  
E Coli ◽  
High Prevalence

Background and Aim: Escherichia coli is the cause of avian colibacillosis, a significant threat to the poultry industry and public health. Thus, this study investigated the prevalence of E. coli in diseased chicken broilers, pathological effects of these bacteria, and interleukin (IL) gene expression of different serotypes of E. coli (O78, O26, O44, and O55) on experimentally infected chickens. Materials and Methods: A total of 295 organ samples (liver, lungs, heart, and spleen) from 59 diseased broiler chickens were used for conventional identification of E. coli. Chickens were orally infected with one of the following E. coli serotypes (O78, O26, O44, or O55) and examined for clinical signs, mortality, macroscopic and microscopic lesions, and IL gene expression using real-time quantitative polymerase chain reaction. Results: E. coli was isolated from 53.2% of broiler chicken organs with a high prevalence in lungs (26.1%). The most prevalent serotypes were O78, O26, O44, O55, O157, and O127 prevalence of 27.8, 22.2, 16.7, 16.7, 5.6, and 5.6%, respectively. In the experimental design, five groups (G1-G5) of birds were established. G1 served as the negative control group, while G2-G5 were challenged orally with E. coli O78, O26, O55, or O44, respectively. Chickens infected with E. coli O78 or O26 showed significant clinical signs in comparison to the other infected birds. Mortality (13.3%) was only observed in birds infected with E. coli O78. Necropsy of dead birds after E. coli O78 infection showed pericarditis, enteritis, airsacculitis, and liver and lung congestion. More severe histopathological changes were observed in intestines, spleen, liver, and lung from chickens infected with either E. coli O78 or O26 than for birds infected with other serotypes. On the 2nd day post-infection, E. coli challenge, particularly with E. coli O78, displayed significantly upregulated levels of ileal IL-6 and IL-8, but ileal IL-10 level tended to be downregulated in comparison to the control group. Conclusion: This study assessed the application of cytokines as therapeutic agents against infectious diseases, particularly colibacillosis.

Fasting-induced changes in ECL cell gene expression

2007 ◽  
Vol 31 (2) ◽  
pp. 183-192 ◽  
Author(s):  
Nils W. G. Lambrecht ◽  
Iskandar Yakubov ◽  
George Sachs
Keyword(s):  
Gene Expression ◽  
Protein Content ◽  
Quantitative Polymerase Chain Reaction ◽  
Cell Types ◽  
Mucosal Cell ◽  
Ecl Cells ◽  
Ecl Cell ◽  
Histamine Uptake ◽  
Cell Gene Expression ◽  
Fasted Rats

Gastric enterochromaffin-like (ECL) cells release histamine in response to food because of elevation of gastrin and neural release of pituitary adenylate cyclase-activating peptide (PACAP). Acid secretion is at a basal level in the absence of food but is rapidly stimulated with feeding. Rats fasted for 24 h showed a significant decrease of mucosal histamine despite steady-state expression of the histamine-synthesizing enzyme histamine decarboxylase (HDC). Comparative transcriptomal analysis using gene expression oligonucleotide microarrays of 95% pure ECL cells from fed and 24-h fasted rats, thereby eliminating mRNA contamination from other gastric mucosal cell types, identified significantly increased gene expression of the enzymes histidase and urocanase catabolizing the HDC substrate l-histidine but significantly decreased expression of the cellular l-histidine uptake transporter SN2 and of the vesicular monoamine transporter 2 (VMAT-2) responsible for histamine uptake into secretory vesicles. This was confirmed by reverse transcriptase-quantitative polymerase chain reaction of gastric fundic mucosal samples from fed and 24-h fasted rats. The decrease of VMAT-2 gene expression was also shown by a decrease in VMAT-2 protein content in protein extracts from fed and 24-h fasted rats compared with equal amounts of HDC protein and Na-K-ATPase α1-subunit protein content. These results indicate that rat gastric ECL cells regulate their histamine content during 24-h fasting not by a change in HDC gene or protein expression but by regulation of substrate concentration for HDC and a decreased histamine secretory pool.

Curcumin Affects Adipose Tissue-Derived Mesenchymal Stem Cell Aging Through TERT Gene Expression

Drug Research ◽  
2017 ◽  
Vol 68 (04) ◽  
pp. 213-221 ◽  
Author(s):  
Samaneh Pirmoradi ◽  
Ezzatollah Fathi ◽  
Raheleh Farahzadi ◽  
Younes Pilehvar-Soltanahmadi ◽  
Nosratollah Zarghami
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Adipose Tissue ◽  
Free Radicals ◽  
Population Doubling ◽  
Cell Aging ◽  
Control Group ◽  
Tert Gene ◽  
Rat Adipose Tissue ◽  
Tert Expression

AbstractAging and losing cell survival is one of the main problems in cell therapy. Aging of Mesenchymal Stem Cells (MSCs) is associated with a rise in intracellular reactive oxygen species, decrease in telomerase reverse transcriptase (TERT) expression and finally eroded telomere ends. Given that the production of free radicals in the aging process is effective, the use of antioxidants can help in scavenging free radicals and prevent the aging of cells. The aim of this study is to evaluate the effects of curcumin on proliferation, aging and TERT expression of rat adipose tissue-derived stem cells (rADSC). rADSCs were isolated from inguinal rat adipose tissue and their viabilities were assessed by MTT after exposure to different concentrations of curcumin. Flow-cytometry was performed for investigating the cell surface markers. Adipogenic and osteogenic differentiation were carried out to evaluate the pluripotency of rADSCs. Telomerase expression and percentage of senescent cells were evaluated using real-time PCR and senescence-associated β-galactosidase activity, respectively. The results demonstrated significant proliferation of rADSCs after 48 h treatment with 1 and 5 µM curcumin. Additionally, these concentrations could significantly reduce the population doubling time and aging of rADSCs at different passages. The findings of SA-ß-gal staining showed that curcumin significantly decreased the number of senescent cells in the 5 and 7 cell passages. Moreover, expression levels of TERT increased in the presence of 1 and 5 µM curcumin than control group (P<0.001). As a conclusion, curcumin may be a good candidate to improve lifespan of rADSCs through promoting TERT gene expression.

scholarly journals Evaluation of Ginkgo biloba and Flunixin in BIM Gene Expression and Viability of Ovarian Cancer Cell A2780s

2021 ◽  
Vol 12 (3) ◽  
Author(s):  
Farnoosh Soleamani ◽  
Elham Salehi ◽  
Majid Morovati-Sharifabad ◽  
Fatemeh Sarkargar ◽  
Gholamhosein Pourghanbari
Keyword(s):  
Gene Expression ◽  
Ovarian Cancer ◽  
Real Time ◽  
Ginkgo Biloba ◽  
Gynecologic Cancer ◽  
Cell Aging ◽  
Control Group ◽  
Anti Cancer ◽  
Ic50 Values ◽  
Apoptosis Regulation

Background: Ovarian cancer is the deadliest gynecologic cancer. Studies on the therapeutic properties of Ginkgo biloba and flunixin showed that these drugs, singly or in combination with other drugs, have anti-cancer activities. Different genes are involved in apoptosis regulation. The BIM gene is one of the most important regulators of this process. BIM has different roles, including cell cycle regulation, apoptosis induction, deoxyribonucleic acid recombination, chromosomal segregation, and cell aging. Methods: This study evaluated the viability percentage of the A2780s cell line with Ginkgo biloba and flunixin at different concentrations, compared to that of the control group. Then, the half-maximal inhibitory concentration (IC50) values of Ginkgo biloba and flunixin were determined within 24 h. Then, the expression of the BIM gene was evaluated using a real-time polymerase chain reaction (PCR). Results: The IC50 results showed that Ginkgo biloba and flunixin significantly reduced cell life (P < 0.01) depending on time and concentration. The results of real-time PCR showed that cell treatment with Ginkgo biloba and flunixin significantly increased BIM expression. Conclusions: The results of this experiment indicated that BIM gene expression was increased in cancer cells treated with Ginkgo biloba and flunixin, compared to that reported for control cells. Therefore, with further research in the future, these compounds can be used for the development of ovarian anti-cancer drugs.

Regulation of Apoptosis and Oxidative Stress by LMP1 Oncoprotein of Epstein-Barr Virus in Patients with Low Grade B-Cell Leukemic Lymphomas

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 5212-5212
Author(s):  
Panagiotis Theodorou Diamantopoulos ◽  
Katerina Polonyfi ◽  
Nikolaos Spanakis ◽  
Athanassios Galanopoulos ◽  
Georgia Diamantopoulou ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Gene Expression ◽  
B Cell ◽  
Cell Lines ◽  
Peripheral Blood ◽  
Cell Aging ◽  
Control Group ◽  
Low Grade ◽  
Qrt Pcr ◽  
Epstein Barr

Abstract Abstract 5212 Background: In all Epstein-Barr (EBV)-associated malignancies, the virus displays a latency program of infection and a restricted pattern of gene expression. Among the products of these genes, latent membrane protein 1 (LMP1) is a potent transforming protein with several different roles. LMP1 has been shown in cell lines to stimulate apoptosis. Survivin, a member of the inhibitor of apoptosis (IAP) family, is an important regulator of the mitochondrial apoptotic pathway, while oxidative stress (OS) is a cellular condition particularly relevant to cell aging. In the present study we enrolled patients with non-EBV-related low grade B-cell lymphoproliferative diseases. The aim was to detect (1) the viral load of EBV-positive patients, (2) the expression of LMP1 oncoprotein, (3) the possible apoptotic properties of LMP1 by correlating the levels of survivin with LMP1 expression, and (4) the levels of oxidative stress in LMP1-positive and negative patients. Patients and Methods: Forty eight Greek patients with EBV-unrelated low grade B-cell leukemic lymphomas, were enrolled in the study (chronic lymphocytic leukemia: 27, marginal zone lymphoma: 12, mantle cell lymphoma: 4, hairy cell leukemia: 2, follicular lymphoma: 2, lymphoplasmacytic lymphoma: 1). The majority of patients (61.2%) were treatment-naïve, while the rest had not received any treatment for at least 6 months. DNA from peripheral blood was tested by quantitative real time (qRT) PCR for the EBV-R gene. RNA from EBV-positive patients was examined by RT-PCR and qRT PCR for LMP-1, while using qRT PCR we measured survivin expression in all patients. Densitometric analysis (DA) was used for semi-quantification of the survivin gene expression. The results were expressed relative to the expression of ABL housekeeping gene. The control group included 30 EBV-negative healthy adults. Oxidative stress was measured in the serum of all patients using the PerOx (TOS/TOC) Kit, by Immunodiagnostik. Non parametric methods (Mann-Whitney test) were used for statistical analysis of the results. Results: Twenty five (25) men and 23 women, with a median age of 74 (51–87 years old) were studied. EBV positivity was detected in 19/48 (39.6%) patients, and LMP1 was expressed in 13/19 (68.4%) EBV-positive patients. Survivin levels were lower in LMP1-positive patients vs LMP1-negative patients (2-tailed p=0.009). The oxidative stress was lower (261.4 μmol/L) in LMP1-positive patients vs LMP1-negative patients (372.3 μmol/L), (2-tailed p=0.014). Discussion: The literature lacks information about the expression of LMP1 in the peripheral blood of patients with non-EBV-related low grade B-cell leukemic lymphomas. Previous studies in LMP1-positive lymphoma cell lines have shown the apoptotic functions of LMP1 during type II latency. In this study LMP1-positive patients express statistically significant lower levels of survivin vs LMP1-negative patients. This finding is in accordance to the hypothesis that LMP1 oncoprotein can induce apoptosis. LMP1-positive patients had lower levels of oxidative stress compared to LMP1-negative patients. According to our findings, in non-EBV-related lymphomas, LMP1 may increase apoptosis and decrease the levels of oxidative stress. Disclosures: No relevant conflicts of interest to declare.

scholarly journals MEASUREMENT OF DNA DAMAGE, OXIDATIVE STRESS, AND GENE EXPRESSION OF β-CATENIN AND P53 GENES IN LIVER AND BRAIN OF MALE MICE RECEIVING MONOSODIUM L-GLUTAMATE MONOHYDRATE

2020 ◽  
pp. 127-132
Author(s):  
NOHA IBRAHIM SAID SALEM ◽  
HANAN R.H. MOHAMED ◽  
AREEG MOHAMED ABD-ELRAZEK
Keyword(s):  
Oxidative Stress ◽  
Gene Expression ◽  
Dna Damage ◽  
Oxidative Dna Damage ◽  
Quantitative Polymerase Chain Reaction ◽  
Significant Decline ◽  
Control Group ◽  
Group I ◽  
Significant Difference ◽  
P53 Genes

Introduction: Monosodium L-glutamate (MSG) monohydrate is a widespread nutritional additive and flavoring agent frequently consumed all over the world. In this study, we investigate the action of daily oral intake of MSG monohydrate in vivo using mammalian systems. Methods: Mice divided as follows: Group I (normal control), Group II, and Group III treated with MSG for 2 and 4 weeks, respectively. Brain and liver dissected out for the detection of fragmented DNA, DNA damage, and assay of oxidative stress markers. Moreover, expression levels of ß-Cat and p53 genes were measured by a real-time quantitative polymerase chain reaction. Results: The results showed a significant difference in MSG-treated group at the 2-time intervals than the control one regarding parameters of oxidative stress, and these were accompanied by a significant decline in glutathione (GSH) and a ratio of oxidized and reduced GSH in both tissues. Significant elevation of laddered DNA and oxidative DNA damage was observed in groups treated with MSG. In addition, a significant decline in gene expression of ß-Catenin in liver and brain tissues with elevations in the gene expression of p53 in the brain. Furthermore, the p53 gene in liver tissue was significantly upregulated in mice administered MSG for 15 days and was downregulated after 30 days of MSG intake compared with the control. Conclusion: According to our results, oral consumption of MSG leads to oxidative stress-mediated DNA damage and apoptosis.

scholarly journals Reductive regulation of BECN1 gene in adult Egyptian patients with do novo AML

2020 ◽  
Vol 21 (1) ◽  
Author(s):  
Manal Fawzy Ghozlan ◽  
Botheina Ahmed Thabet Farweez ◽  
Nesma Ahmed Safwat ◽  
Noha Bassiouny Hassan ◽  
Walaa Ali Elsalakawy
Keyword(s):  
Gene Expression ◽  
Bone Marrow ◽  
Peripheral Blood ◽  
De Novo ◽  
Quantitative Polymerase Chain Reaction ◽  
Control Group ◽  
Significant Difference ◽  
Egyptian Patients ◽  
Blast Cells ◽  
De Novo Aml

Abstract Background Acute myeloid leukaemia (AML) is a clonal haematopoietic disease characterized by the proliferation of immature blast cells in the bone marrow and peripheral blood. Autophagy is an inherent cellular route by which waste macromolecules are engulfed within autophagosomes prior to their fusion with cytoplasmic lysosomes for degradation. The BECN1 gene encodes the Beclin-1 protein, which regulates autophagy. Few reports have investigated BECN1 gene expression and its value in AML patients. Results This randomized case-control study included 50 newly diagnosed AML patients, in addition to 20 subjects as a control group. BECN1 gene expression was assessed using real-time quantitative polymerase chain reaction (qRT-PCR). The median level of BECN1 gene expression in AML patients was 0.41 (IQR 0.29–1.03) in comparison to 1.12 (IQR 0.93–1.26) in the control group (P = 0.000). Seventy-two percent of AML patients showed reduced BECN1 gene expression, which was highly significantly associated with intermediate and adverse cytogenetic risk. Reduced BECN1 gene expression was associated with older age, higher total leukocyte counts, the presence of peripheral blood blast cells, a higher percentage of bone marrow blast cells, and higher expression of CD34 and CD117. FLT3-ITD mutation was detected in 14 patients (38.9%), all of whom showed reduced BECN1 gene expression (P = 0.006). BECN1 gene expression was also reduced in non-responder AML patients, with a highly statistically significant difference (P = 0.002). Conclusion A reduction in BECN1 gene expression might indicate a poor prognosis in adult Egyptian patients with de novo AML. Decreased BECN1 gene expression is associated with a higher risk of resistance to treatment. Targeting autophagy pathways may help in the treatment of AML patients.

scholarly journals RUNX1 gene expression in Egyptian acute myeloid leukemia patients: may it have therapeutic implications?

2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Fadwa Said ◽  
Roxan E. Shafik ◽  
Naglaa M. Hassan
Keyword(s):  
Gene Expression ◽  
Acute Myeloid Leukemia ◽  
Myeloid Leukemia ◽  
Quantitative Polymerase Chain Reaction ◽  
Gene Expression Level ◽  
Expression Level ◽  
Control Group ◽  
Therapeutic Implications ◽  
Number Of Patients ◽  
Acute Myeloid

Abstract Background Acute myeloid leukemia represents the highest percentage of all adult acute leukemia variants. Runt-related transcription factor1 (RUNX1), a transcription factor with a known tumor suppressor function, was recently reported as a tumor promoter in acute myeloid leukemia (AML). We investigated the role of RUNX1 gene expression level in Egyptian AML patients and delineated its clinical significance. Results We measured RUNX1 gene expression level using reverse transcription-quantitative polymerase chain reaction and found that the RUNX1 gene expression level was significantly higher than the control group (p < 0.001). Patients with FMS-like tyrosine kinase 3 internal tandem duplication (FLT3-ITD) mutations had a higher expression level of RUNX1 (p = 0.023). The male patients expressed a significantly higher level of RUNX1 (p = 0.046). Conclusions The RUNX1 gene is highly expressed in Egyptian AML patients. It has a relation to FLT3-ITD, which may give a clue that patients carrying this mutation may benefit from new treatments that target RUNX1 in the future. Further studies on a larger number of patients with different ethnic groups may give a clearer vision of the therapeutic implications of a new molecular target.

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