Cytokine Gene Polymorphisms in Turkish Patients with Childhood Acute Lymphoblastic Leukemia.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4441-4441
Author(s):  
Kursat Ozdilli ◽  
Fatma Oguz ◽  
Hulya Bilgen ◽  
Yeliz Duvarci ◽  
Halim Issever ◽  
...  
Keyword(s):  
Acute Lymphoblastic Leukemia ◽  
Odds Ratio ◽  
Lymphoblastic Leukemia ◽  
The Other ◽  
Control Group ◽  
Target Cells ◽  
Cytokine Gene ◽  
Chi Square ◽  
Other Hand ◽  
Ifn Γ

Abstract Cytokines are chemical mediators between cells and they bind on their own specific receptors on the target cells. Cytokine gene polymorphism may be implicated in the pathogenesis of infections autoimmun disease and malignancies via their effect on cytokine production and regulation. Leukemic cells proliferate under the influence of cytokines. It is known that acute lymphoblastic leukemia (ALL) which is sensitive to some cytokine gene is one of the hematological malignances derived from lymphoid tissue. Our aim is to determined frequencies of selected cytokines (TNF-α, TGF-β, IL-10, IL-6, IFN-γ) in childhood ALL patients and unrelated healthy control groups. We also investigated weather these polymorphism might contribute to the pathogenesis of ALL. Method: Genotyping was performed by polymerase chain reaction with sequence specific primers (PCR-SSP) using “ One lamda” kit in 44 ALL patients and 50 unrelated healthy individuals. Allelic frequencies were determined 24 polymorphism located within 16 cytokine genes. Genotype frequencies between ALL patients and controls were compared using Chi-Square Yates, Fisher’s Exact Tests and Stepwise Logistic Regression Analysis (SLRA). Results: The frequency of TGF-β(TT/GG) allel (29,5% versus 8%, respectively;in chi-square p=0,007,odds ratio=4,82, 95% confidence interval (CI) 1,49–16,16; in SLRA p=0,002, OR=15,26,95% CI 2,77–84,05), TGF-β(TC/GG) allel (36,4% versus 10%, respectively; in chi-square p=0,002,odds ratio=5,14, 95% CI 1,69–15,59; in SLRA p=0,005,OR=10,21, 95% CI 2,02–51,61), IL-6(GC) allel (38,6% versus 8%, respectively; in chi-square p<0,001; odds ratio 7,24; 95% CI 2,20–23,76; in SLRA p=0,02,OR=6,37, 95% CI 1,22–33,15), IL-10(GCC/ATA) allel (36,4% versus 8%, respectively; in chi-square p=0,001, odds ratio 6,57, 95% CI 1,99–21,64; in SLRA p=0,01,OR=8,04, 95% CI 1,40–46,04), IFN-γ(TA) allel (54,5% versus 10%, respectively; in chi-square p<0,001, odds ratio 10,8, 95% CI 3,60–32,40; in SLRA p<0,001,OR=29,43, 95% CI 5,47–158,35) frequencies were found higher in patients with ALL compared to the control group. On the other hand IFN-γ(TT) allel (20% versus 48%, respectively; p=0,005; odds ratio 0,27; 95% CI 0,11–0,70), IFN-γ(AT)(0% versus 22%, respectively; p=0,001; odds ratio 1,28; 95%CI 1,10–1,48; fisher’s exact=0,001) and IL-6(CA)(0% versus 22%, respectively; in chi-square p<0,001; odds ratio 1,28; 95% CI 1,10–1,28; fisher’s exact=0,001) allel frequencies were found higher in the control group compared to the patients with ALL.TNF-α(GG) allel frequency was higher both in patients with ALL and in the control group (72% versus 85%). As a conclusion higher frequency in TGF-b(TT/GG),TGF-b(TC/GG),IL-6(GC), IL-10(GCC/ATA) and IFN-g(TA) allels may predispose to ALL.On the other hand IFN-g(TT), IFN-g(AT) and IL-6(CA) allels might be the preventive factor for ALL The present study is rather significant that it is the study which assessed the relation of the cytokines in patients with ALL compared to the control group taken from the genetic pool of The Turkish population but larger groups of studies must be done in future.

Cytokine Gene Polymorphisms in Turkish Patients with Childhood Chronic Myeloid Leukemia.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 4368-4368
Author(s):  
Kursat Ozdilli ◽  
Fatma Oguz ◽  
Yeliz Duvarci ◽  
Hulya Bilgen ◽  
Sema Anak ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Chronic Myeloid Leukemia ◽  
Myeloid Leukemia ◽  
The Other ◽  
Fine Tuning ◽  
Control Group ◽  
Cytokine Gene ◽  
Chi Square ◽  
Other Hand ◽  
Genotype Frequencies

Abstract Cytokines are necessary for normal hematopoiesis in the bone marrow and provide a means of fine-tuning bone marrow function in response to stimulation. Several of cytokines generated during both innate and adaptive immune responses stimulate the growth and differantiation of bone marrow progenitor cells. The vast majority of polymorphism found in cytokine genes and their reseptors are located in non coding regions. Cytokine gene polymorphism may be implicated in the pathogenesis of infections, autoimmun disease and malignancies via their effect on cytokine production and regulation. It is known that leukemic cells proliferate under the influence of cytokines. Our aim is to analyze cytokine gene profiles in patients with chronic myeloid leukemia (CML)in order to clarify the pathogenesis of CML. Genomic DNA from 26 CML patients were analysed. Genotyping was performed by polymerase chain reaction with sequence specific primers (PCR-SSP) using “ One lamda” kit in 26 childhood CML patients 60 unrelated healthy individuals. Genotype frequencies between CML patients and controls were compared using Chi-Square Yates, Fisher’s Exact Tests. In this study; 26 CML patients with a mean age of 15,6±5,3 and 60 healthy controls with a mean age of 18,2±5,4 were investigated. In CML patients the frequencies of TGF-b(TC/GG) genotype (in chi-square p=0,01,odds ratio(OR)=3,46, 95% confidence interval (CI) 1,3–9,04) and IL-10 (GCC/ATA) genotype (in chi-square p=0,04,OR=3,3, 95% CI 0,9–11,1 were found higher in patients with CML compared to the control group. On the other hand IL-6 (CC) genotype (in chi-square p=0,012,OR=0,17, 95% CI 0,4–0,76 frequency was found higher in the control group compared to the patients with CML. As a conclusion higher frequency in TGF-b(TC/GG) and IL-10(GCC/ATA) genotype polymorphism were significantly higher in the patients with CML so it may predispose to CML. On the other hand IL-6 (CC) genotype might be the preventive factor for CML. The present study is rather significant that it is the study which assessed the relation of the cytokines in patients with CML compared to the control group taken from the genetic pool of The Turkish population, we believe studies like this will eventually help to understand the pathogenesis of CML and the role cytokines play in CML,but larger groups of studies must be done in future.

Association of CYP2B6 c.516G>T Polymorphism with Acute Lymphoblastic Leukemia Susceptibility

2014 ◽  
Vol 926-930 ◽  
pp. 1073-1076
Author(s):  
Zhong Hai Yuan ◽  
Yi Ju Hou ◽  
Chen Zhao ◽  
Yan Li
Keyword(s):  
Acute Lymphoblastic Leukemia ◽  
Genomic Dna ◽  
Lymphoblastic Leukemia ◽  
Control Group ◽  
Study Group ◽  
Regression Analyses ◽  
Specific Primers ◽  
Blood Leukocytes ◽  
Chi Square ◽  
Chi Square Test

Abstract:Objective: To investigate whether any association exists between genetic polymorphism in CYP2B6 c.516G>T and individual susceptibility to acute lymphoblastic leukemia (ALL). Methods: Our study group consisted of 96 ALL patients(T-ALL 17 cases, B-ALL 79 cases) and 348 unrelated healthy newborn volunteers as a control group. Genomic DNA was extracted from peripheral blood and cord blood leukocytes. We genotyped CYP2B6 c.516G>T polymorphism by use of PCR with sequence-specific primers (PCR-SSP). The data were analyzed statistically using chi-square and logistic regression analyses. Results: The frequencies of GG genotype were 74.14%, 57.29%, 29.41% and 63.29%, and GT genotype were 23.85%, 37.50%, 64.71% and 31.65%, and TT genotype were 2.01%, 5.21%, 5.88% and 5.06% in control group, ALL, T-ALL, and B-ALL cases, respectively. Chi-square test showed a significant correlation between the CYP2B6 c.516G>T polymorphism GT genotype and ALL patients (OR=2.035, 95%CI=1.249-3.313, P=0.004); and T-ALL patients (OR=6.839, 95%CI=2.309-20.252, P=0.000); whereas and B-ALL patients (OR=1.554, 95%CI=0.906-2.667, P=0.108). Conclusions: This study revealed the CYP2B6 c.516GT genotype may be a risk factor to the development of ALL, especially T-ALL.

Effects Of Metformin Addition To a Standard Chemotherapy Regimen For The Treatment Of Adult Acute Lymphoblastic Leukemia. An In Vitro and In Vivo study

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 5024-5024
Author(s):  
Christian Omar Ramos Peñafiel ◽  
Irma Olarte Carrillo ◽  
Jorge Zamora Domínguez ◽  
Humberto Baldemar Castellanos Sinco ◽  
Etta Rozen Fuller ◽  
...  
Keyword(s):  
Regression Analysis ◽  
Acute Lymphoblastic Leukemia ◽  
Odds Ratio ◽  
Cox Regression ◽  
Lymphoblastic Leukemia ◽  
Control Group ◽  
Cox Regression Analysis ◽  
Adult Acute Lymphoblastic Leukemia

Abstract Antitumor effects of metformin are widely known. Unfortunately their use in the treatment of acute lymphoblastic leukemia is limited. Study design The study consisted of two stages Phase 1: Cell assay adding metformin (40mM) assessing the viability and cell cycle in the cell line molt 4 Phase 2: Pilot study adding Metformin (850mg three time day) during the pre-treatment with steroids and the induction remission phase versus a control group (2:1 randomization) Statistical analysis Chi-square analysis was used to corroborate the hypothesis. The odds ratio was calculated for both, the absence of Good steroid response and refractory leukemia. Cox regression analysis and Kaplan-Meier curves were used for the survival analysis Results Celular assay. Metformin inhibited cell viability at 120hours reducing the percentage of cells in phase S. Clinical assay. 151 patients were studied, 44 (29.1%) on Metformin arm, of these 59.1% ( n=26) archived a GSR compared with the control group (26.2%, n=28). A greater number of complete remission were presented in Metformin arm (81.8% versus 57.9%) shown to be a protective factor for GSR and complete remissions (odds ratio ; 0.2454 y 0.3062 respectively). In the Cox regression analysis, Metformin significantly impact in the global survival (p=0,012,95% IC) versus the other variables studied. Conclusions Metformin is useful both in vivo and in vitro treatment of adult acute lymphoblastic leukemia. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Pancytopenia – (?) An obstacle in the diagnosis and outcome of pediatric acute lymphoblastic leukemia

2015 ◽  
Vol 04 (02) ◽  
pp. 068-071 ◽  
Author(s):  
Shruti Raja ◽  
Febe Renjitha Suman ◽  
Julius Xavier Scott ◽  
M. S. Latha ◽  
Aruna Rajenderan ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Acute Lymphoblastic Leukemia ◽  
Lymphoblastic Leukemia ◽  
Cross Sectional Study ◽  
Control Group ◽  
Clinical Expertise ◽  
Chi Square ◽  
Cross Sectional ◽  
Significant Difference ◽  
Chi Square Test

Abstract Context: Acute lymphoblastic leukemia (ALL) ranks first among pediatric malignancies. 8-12% of ALL present with pancytopenia and 2% with hypocellular marrow a. Diagnosis of ALL in the background of pancytopenia and aplastic bone marrow is difficult. Aims: This study was aimed to compare the clinicopathologic, genetic, and outcome of paediatric ALL patients with and without pancytopenia. Settings and Design: This is a retrospective cross-sectional study. Susbjects and Methods: The study included all ALL patients presenting with pancytopenia. The control group included equal number of randomly selected patients with ALL without pancytopenia treated during the same period. Ethics committee approved this study. The demographic, laboratory, and treatment-related details were retrieved from the records and entered in an Excel sheet. Statistical Analysis Used: Data was analyzed with Chi-square test with IBM SPSS statistics 16 software. Results: Diagnosis by peripheral smear is significantly lower (P = 0.015) in comparison with the control group. There is no significant difference in diagnosis between the groups by bone marrow aspirate (P = 0.731) and biopsy (P = 0.849). The diagnosis of leukemia is misdiagnosed as hypo cellular/aplastic marrow in 10% of the pancytopenic patients. Flow cytometry yielded the diagnosis in all the pancytopenic patients. Though cytogenetic abnormalities are more common in pancytopenic group, it is not statistically significant (P = 0.106). There is no significant difference in treatment outcome between the groups (P = 0.0827%). Conclusions: Clinical expertise is highly essential to evaluate a case of pancytopenia to diagnose leukemia. Pancytopenia is an obstacle in the diagnosis of ALL without immunophenotyping. There is no significant difference in the outcome between the two groups.

scholarly journals Ghrelin and orexin levels in infertile male: evaluation of effects on varicocele pathophysiology, relationship seminal and hormonal parameter

2020 ◽  
Vol 0 (0) ◽  
Author(s):  
Üçler Kısa ◽  
M. Murad Başar ◽  
Timuçin Şipal ◽  
Özlem Doğan Ceylan
Keyword(s):  
The Other ◽  
Control Group ◽  
Infertile Male ◽  
Seminal Parameters ◽  
Other Hand ◽  
Healthy Control ◽  
Regulatory Effects ◽  
Serum Ghrelin ◽  
Infertile Patients ◽  
Group 1

AbstractObjectiveThe aim of the present study was to investigate serum ghrelin and orexin levels in patients with varicocele and compare these levels with idiopathic infertile male and healthy control cases.MethodsThis study enrolled 24 men with varicocele, 24 males having idiopathic infertility, and 21 fertile men as the control group. Hormonal analyses, ghrelin and orexin levels were measured samples. Semen was analyzed after 3 and 5 days of sexual abstinence.ResultsSerum ghrelin levels were statistically different among the three groups (p=0.015), and it was due to a statistically lower level in group-1 than the level in the control cases (p=0.012). On the other hand, serum orexin levels were lower than healthy subjects in infertile groups with/without varicocele, but there was no difference (p=0.685) among three groups. Serum ghrelin level showed a negative and significant correlation only with sperm motility (r=−0.646, p=0.022), there was no correlation with other parameters. On the other hand, serum orexin levels did not show a significant correlation with seminal parameters.ConclusionBoth new investigated peptides ghrelin and orexin have regulatory effects on testicular function. However, ghrelin has a more obvious and complex effect on spermatogenesis. Impaired seminal parameters, especially motility was associated with increased serum ghrelin levels in infertile patients, especially with varicocele.

scholarly journals The Effect of Geogebra on Students’ Conceptual and Procedural Knowledge: The Case of Applications of Derivative

2017 ◽  
Vol 7 (2) ◽  
pp. 67 ◽  
Author(s):  
Mehmet Fatih Ocal
Keyword(s):  
Conceptual Knowledge ◽  
Teaching And Learning ◽  
Procedural Knowledge ◽  
Dynamic Geometry ◽  
The Other ◽  
Control Group ◽  
Other Hand ◽  
Significant Difference ◽  
Before And After ◽  
Post Test

Integrating the properties of computer algebra systems and dynamic geometry environments, Geogebra became an effective and powerful tool for teaching and learning mathematics. One of the reasons that teachers use Geogebra in mathematics classrooms is to make students learn mathematics meaningfully and conceptually. From this perspective, the purpose of this study was to investigate whether instruction with Geogebra has effect on students’ achievements regarding their conceptual and procedural knowledge on the applications of derivative subject. This study adopted the quantitative approach with pre-test post-test control group true experimental design. The participants were composed of two calculus classrooms involving 31 and 24 students, respectively. The experimental group with 31 students received instruction with Geogebra while the control group received traditional instruction in learning the applications of derivative. Independent samples t-test was used in the analysis of the data gathered from students’ responses to Applications of Derivative Test which was subjected to them before and after teaching processes. The findings indicated that instruction with Geogebra had positive effect on students’ scores regarding conceptual knowledge and their overall scores. On the other hand, there was no significant difference between experimental and control group students’ scores regarding procedural knowledge. It could be concluded that students in both groups were focused on procedural knowledge to be successful in learning calculus subjects including applications of derivative in both groups. On the other hand, instruction with Geogebra supported students’ learning these subjects meaningfully and conceptually.

scholarly journals Identification and Validation of Serum Autoantibodies in Children with B-cell Acute Lymphoblastic Leukemia by Serological Proteome Analysis

2021 ◽  
Author(s):  
Runhong Yu ◽  
Shiwei Yang ◽  
Yufeng Liu ◽  
Zunmin Zhu
Keyword(s):  
Acute Lymphoblastic Leukemia ◽  
Western Blot ◽  
B Cell ◽  
Lymphoblastic Leukemia ◽  
Immunohistochemical Analysis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Healthy Children ◽  
Expression Levels ◽  
Cell Acute Lymphoblastic Leukemia

Abstract Purpose: Study was by intention to screen serum autoantibodies that may contribute to the early detection of B-cell acute lymphoblastic leukemia (B-ALL) in children.Patients and methods: The total protein from three pooled B-ALL cell lines(NALM-6, REH and BALL-1 cells) was separated using two-dimensional gel electrophoresis(2-DE), which was followed by Western blot by mixed serum from B-ALL patients (n=20) or healthy children(n=20). We obtained and analyzed the images of 2-D gel and Western blot by PDQuest software,and then identify the spots of immune responses in B-ALL samples compared with those in control samples.The proteins from spots were identified using mass spectrometry (MS). The autoantibodies against α-enolase and voltage-dependent anion-selective channel protein 1(VDAC1) were further validated on the use of enzyme-linked immunosorbent assay(ELISA). The protein expression levels of the candidate antigens α-enolase and VDAC1 in B-ALL were thoroughly studied by immunohistochemical analysis.Results: Six protein dots were identified with MS as Aconitase,apoptosis-inducing factor(AIF),dihydrolipoamide dehydrogenase(DLD), α-enolase,medium-chain acyl-CoA dehydrogenase(MCAD) and VDAC 1.The frequencies of autoantibodies against α-enolase and VDAC1 in children with B-ALL were 27% and 23%, respectively, which were significantly higher than those in normal controls(4% and 0). Immunohistochemical analysis showed the expression of α-enolase and VDAC1 was positive in 95% and 85% of B-ALL patients, respectively, but negative expression levels were showed in the control group. Conclusion: This study incidates that α-enolase and VDAC1 may be the antigen associated with B-ALL .α-enolase and VDAC1 autoantibodies may develop into potential serological markers of B-ALL in children.Other proteins also need to be confirmed in a large number of serum samples.

scholarly journals To determine how the effect of station technique on students’ achievement in astronomy subjectsAstronomi konularında istasyon tekniğinin öğrencilerin akademik başarısına nasıl etki ettiğini belirlemek

2017 ◽  
Vol 14 (4) ◽  
pp. 4561 ◽  
Author(s):  
Hamza Albayrak ◽  
Paşa Yalçın ◽  
Sema Altun Yalçın
Keyword(s):  
Quantitative Methods ◽  
T Test ◽  
The Other ◽  
Control Group ◽  
Science Lesson ◽  
Test Model ◽  
Meaningful Difference ◽  
Control Groups ◽  
Other Hand ◽  
Post Test

In this research, it was aimed to determine the effect of learning stations designed for astronomy subjects on the academic achievement of students and give an alternative method for teaching astronomy in the literature. The mixed approach in which mixed quantitative and qualitative methods were used together was preferred in the study. Among the quantitative methods, the pre-test and post-test model of the semi-experimental method was used; in addition, the protocols were applied about the method, which was used. The research was carried out with 98 seventh class students, who were under education in a secondary school located in the Eastern Anatolia Region of Turkey in the education year of 2015-2016. The quantitative data were analysed via the Astronomy Achievement Test (AAT) developed by the researchers and had 0,795 Cronbach Alpha reliability coefficient. The astronomy subjects in science lesson of the seventh class were studied with the experimental group through the learning stations; on the other hand, they were studied with the students in the control group through the activities stated in the textbook prepared by the Ministry of Education. The AAT was applied for the experimental, control groups using the pre- and post-tests, and data were analysed with the t-test. Meaningful difference was determined in AAT pre-test average scores of experimental and control groups; on the other hand, a meaningful difference was determined between the groups according to the independent samples t-test results applied with the average scores of the post-test(p<0.05). As a result of the analysis of the data gathered with the Station Observation Form (SOF), developed for the research, it was found out that the students recognise the learning stations as a technique of useful, entertaining, engaging, and easy-to-learn that enable them to learn astronomy topics effectively.Extended English abstract is in the end of Full Text PDF (TURKISH) file. ÖzetBu araştırmada, astronomi konuları için tasarlanan öğrenme istasyonlarının öğrencilerin akademik başarılarına etkisini belirlemek ve literatüre astronomi öğretimi için alternatif bir yöntem sunmak amaçlanmıştır.  Araştırmada karma nicel ve nitel yöntemlerin birlikte kullanıldığı karma yaklaşım tercih edilmiştir. Nicel yöntemlerden yarı deneysel yöntemin ön test- son test modeli kullanılmış olup ayrıca uygulanan yöntem hakkında öğrenci görüşmeleri yapılmıştır. Araştırma 2015- 2016 eğitim öğretim döneminde Doğu Anadolu Bölgesinde yer alan bir ortaokulda öğretim gören 98 yedinci sınıf öğrenci ile yürütülmüştür.  Nicel veriler için araştırmacılar tarafından geliştirilen ve cronbac’h alpha güvenlik katsayısı 0.795 olan Astronomi Başarı Testi (ABT) kullanılmıştır.  Yedinci sınıf fen bilimleri dersi içerisindeki astronomi konuları deney grubu ile öğrenme istasyonları yardımıyla, kontrol grubundaki öğrencilerle MEB’in ön gördüğü ders kitabında yer alan etkinlikler takip edilerek işlenmiştir. ABT deney ve kontrol gruplarına ön test- son test olarak uygulanarak t testi ile analiz edilmiştir. Araştırmada deney ve kontrol gruplarının ABT ön test ortalama puanları arasında anlamlı bir farklılık belirlenmişken, son test puan ortalamaları ile yapılan bağımsız örnekler t testi sonuçlarına göre gruplar arasında anlamlı bir farklılık belirlenmiştir (p<0.05). Araştırma için geliştirilen İstasyon Gözlem Formu (İGF) ile elde edilen verilerinin analizi sonucunda öğrenciler öğrenme istasyonlarını, astronomi konuları için etkili öğrenmeyi sağlayan, faydalı, eğlenceli, ilgi çekici ve kolay öğrenmeyi destekleyen bir teknik olarak gördükleri bulunmuştur.

scholarly journals Transcriptional, Mitochondrial Activity, and Viability of Egyptian Buffalo’s Granulosa Cells In Vitro Cultured under Heat Elevation

2021 ◽  
Vol 11 (2) ◽  
pp. 193-201
Author(s):  
Nasser Ghanem ◽  
Marwa Said Faheem ◽  
Romysa Samy ◽  
Ashraf Hesham Barkawi
Keyword(s):  
Heat Stress ◽  
Granulosa Cells ◽  
Transcriptional Activity ◽  
The Other ◽  
Control Group ◽  
Mitochondrial Activity ◽  
Fluorescent Intensity ◽  
Stressed Group ◽  
Other Hand

It is documented that heat stress caused impairment on the reproductive performance of dairy animals. However, there are few reports that have focused on the molecular and intracellular responses of in vitro cultured buffalo granulosa cells during heat elevation. The present study was conducted to investigate the effect of heat elevation during in vitro culture of buffalo granulosa cells on their viability, quality, mitochondrial activity, and transcriptional activity. Granulosa cells were harvested after aspiration of cumulus-oocytes complexes that were collected from abattoir ovaries. The granulosa cells were cultured in vitro either at a normal physiological temperature suitable for oocyte maturation and embryo development (38.5°C) or exposed to the elevated temperature of 40.5°C on day 3 of culture (the first two days were for confluence) for two hours of culture then continued at 38.5°C up to day 7 of culture. The viability of granulosa cells was measured using trypan blue and quality was estimated by measuring the level of intracellular reactive oxygen species (ROS) on day 7. Moreover, metabolic activity was performed by measuring the fluorescent intensity of mitochondria. Moreover, transcriptional activity was done by profiling four selected candidate genes using quantitative real-time PCR. The results indicated that the granulosa cells viability rate significantly decreased in the heat stress group (25.1 ± 3.7), compared to the control group (36.6 ± 5.3) on confluence day (day 3). In addition, the viability rate on the last day of culture (day 7) decreased in heat stress, compared to control (83.7 ± 4.5 and 97.4 ± 0.4, respectively). On the other hand, there was a nonsignificant difference in ROS profile between the control (21.7*104 ± 1.3) and the heat-stressed group (15.7 ± 0.7) on day 7 of culture. However, the mitochondrial fluorescent intensity was higher in the control (21.9 ± 1.9) than in the heat-stressed group (15.4 ± 0.8) on day 7 of culture. The expression of cellular defense (HSF1) and apoptosis-inducing gene (P53) were significantly up-regulated in granulosa cells exposed to heat elevation, compared to the control group. On the other hand, the steroidogenesis-regulating gene (StAR) was down-regulated in granulosa cells cultured under heat shock, compared to the control group. In conclusion, heat stress reduced the viability of granulosa cells by inducing the expression of an apoptosis-related gene (P53) and compromised expression of genes regulating the steroid biosynthesis, which resulted in up-regulation of cell defense gene (HSF1) in an attempt to ameliorate the deleterious effect of heat stress on the biological activity of the granulosa cells.

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