Hematopathology
Shaghayegh Rostami Yasuj; Narges Obeidi; Gholamreza Khamisipour; Zeynab Gharehdaghi; Zivar Zangeneh
Abstract
Background & Objective: Acute lymphoblastic leukemia (ALL) is a malignant disease that arises from various mutations in B or T-lymphoid progenitors. MicroRNAs (miRNAs) regulate gene expression by binding to the 3' untranslated region of protein-coding genes. Dysregulation of miRNA expression ...
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Background & Objective: Acute lymphoblastic leukemia (ALL) is a malignant disease that arises from various mutations in B or T-lymphoid progenitors. MicroRNAs (miRNAs) regulate gene expression by binding to the 3' untranslated region of protein-coding genes. Dysregulation of miRNA expression may result in the development of cancerous phenotypes. Therefore, for the first time in this field, the present study aims to investigate the effect of overexpression of miR-506 in Jurkat (acute T cell leukemia) cell line. Methods: In this study, Jurkat cell lines were cultured in RPMI-1640 medium. Next, miR-506 was transfected with concentrations of 50 and 100 nM with Lipofectamine 2000. The accuracy of the transfection was confirmed by the transfection of siRNA conjugated with FITC. 48 h after transfection, the cells were prepared for other tests (flow cytometry, MTT assay, and RNA extraction). The expression level of miR-506 in the cells was analyzed using the quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR). Finally, SPSS 21 software was used for the data analysis. Results: According to our results, the viability of cells in concentrations of 50 and 100 nM was significantly higher than the control group. By overexpression of miR-506, the expressions of pro-apoptotic genes (p53, p21) and anti-apoptotic gene B-cell lymphoma-2 (BCL-2) are decreased and increased, respectively. Conclusion: This study showed that miR-506 may function as an oncogenic miRNA in the T- ALL cell line. In conclusion, overexpression of miR-506 leads to an increase in viable cancer cells.
Oral Pathology
Pooja Sharma; Anjali Narwal; Mala Kamboj
Abstract
Background & Objective: Cell population and turnover are controlled by a balance between cell proliferation and apoptosis. Detection of apoptosis in oral cancer contributes to its better prognosis and improved management. This study aimed to quantify apoptotic cells in leukoplakia and oral squamous ...
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Background & Objective: Cell population and turnover are controlled by a balance between cell proliferation and apoptosis. Detection of apoptosis in oral cancer contributes to its better prognosis and improved management. This study aimed to quantify apoptotic cells in leukoplakia and oral squamous cell carcinoma (OSCC) using methyl green-pyronin (MGP) and hematoxylin and eosin (H & E) staining. Methods: The sample included a total of 130 subjects (comprising 108 males and 22 females). Formalin fixed and paraffin embedded tissues were used and categorized into three groups of normal oral mucosa (n=10), leukoplakia with dysplasia (n=60), and OSCC (n=60). The number of apoptotic cells and apoptotic index (AI) were calculated after staining with MGP and routine H & E stained slides. Results: MGP stained the condensed chromatin of apoptotic cells. Statistically significant difference (P≤0.001) was observed among various study groups in terms of numbers of AI and apoptotic cells. Also, AI increased with increasing grades of dysplasia, and it was the highest in well differentiated OSCC. Results were statistically significant in both H & E and MGP stained sections (P≤0.001). A good correlation was found between MGP and H & E staining results. Conclusion: MGP is more specific and can lead to intense staining for chromatin in apoptotic cells. Accordingly, it can provide a good alternative to H&E in identifying apoptotic cells.
Molecular Pathology
Armin Attar; Mohsen Khosravi Maharlooei; Mohammad Nazarnia; Ahmad Hosseini; Zohre Bajalli; Yalda Sadat Moeini; Ahmad Monabati; Fatemeh Amirmoezi; Mansooreh Jaberipour; Mojtaba Habibagahi
Abstract
Background & Objective: It is not clear whether activated lymphocytes of patients with systemic lupus erythematosus (SLE) are more proliferative or less apoptotic. We aimed to delineate potential differences between B and T cells of SLE patients compared to healthy controls regarding the telomerase ...
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Background & Objective: It is not clear whether activated lymphocytes of patients with systemic lupus erythematosus (SLE) are more proliferative or less apoptotic. We aimed to delineate potential differences between B and T cells of SLE patients compared to healthy controls regarding the telomerase activity and apoptosis status. Methods: In this cross-sectional case control study, Blood samples were taken from 10 SLE patients and 10 healthy controls. B and T cells were separated using magnetic cell sorting system. Telomeric repeat amplification protocol (TRAP) assay and real-time PCR were used to determine the telomerase activity and the expression of alternatively spliced variants. Result: Four patients under treatment showed significant telomerase activity in their T cells. Four of the newly diagnosed patients showed telomerase activity in their B cells (20% of all patients and 40% of new onset patients). There was no specific pattern of human telomerase reverse transcriptase variant expression within the patients’ lymphocytes. A significantly reduced expression of Bcl-2 was detected in B cells (P=0.018) and a trend toward lower Bcl-2 expression in T cells was seen in SLE patients compared to healthy controls. Conclusion: Although not definitive, our results may suggest that B cells may have more active roles during the earlier phases of the disease attack, while T cells take over when the disease reaches its chronic stages.
Zeeba Jairajpuri; Rekha Ghai; Sumita Saluja; Sujala Kapur; K.T Bhowmick
Abstract
Background & objective: The current study aimed to perform an immunohistochemical analysis of patterns of apoptotic and cell proliferative related protein expression in different histological grades and immune phenotypes of malignant lymphomas and other lymphoproliferative disorders Methods:This ...
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Background & objective: The current study aimed to perform an immunohistochemical analysis of patterns of apoptotic and cell proliferative related protein expression in different histological grades and immune phenotypes of malignant lymphomas and other lymphoproliferative disorders Methods:This observational study was carried on 60lymph node biopsies of lymphoproliferative disorders. The biopsies were analyzed histologically and immunohistochemically. Results:A total of 60 lymph node biopsies were included in the study, of which 81.6% were of malignant lympho-proliferative lesions. The majority of the biopsies were B-cell (66%) and were grouped in the intermediate grade. Bax and BCL-2 protein expression was presented by percentage of immune positive neoplastic cells per 10fields and graded on a scale of 1 to4. A Bcl-2, Bax Protein Ratio (BBPR) was determined for each case by dividing the estimated Bcl-2 protein (percentage of Bcl-2 positive cells x Bcl-2 staining intensity) by the estimated Bax protein (percentage of Bax positive cells x Bax immunostaining intensity). The mean BBPR was found to be significantly higher in indolent lymphomas (2.64 ± 1.3) as compared to aggressive lymphomas (0.47 ± 0.9) (P<0.01). The expression of P53 and PCNA in 35 biopsies of Non Hodgkin Lymphomas (NHL) was found to increase from low to high grade tumors. Conclusions: A significant correlation was found between BBPR and predicted biological behavior of indolent and aggressive lymphomas. This indicates the important role of Bcl-2 and Bax in biological behavior of lymphomas. Furthermore, P53 and PCNA expression were found to increase from low to high-grade tumors suggesting their prognostic value in NHL.
Ali Reza Khalatbary; Behrooz Mohammadnegad; Ghazaleh Goudarzi; Ali Fazlollahpour Balef
Abstract
Background: There is accumulating evidence that a polyphenol present in olive oil, oleuropein, has antioxidant, anti-inflammatory and anti-apoptotic effects. This study aimed at determining the anti-apoptotic effect of Oleuropein (Ole) on dexamethasone-induced apoptosis of mouse thymocytes. Method: Mice ...
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Background: There is accumulating evidence that a polyphenol present in olive oil, oleuropein, has antioxidant, anti-inflammatory and anti-apoptotic effects. This study aimed at determining the anti-apoptotic effect of Oleuropein (Ole) on dexamethasone-induced apoptosis of mouse thymocytes. Method: Mice were randomly divided to four groups as follow: Dexamethasone (Dex)-treated group (20 mg/kg; single dose), Ole-treated group (20 mg/kg per day), Dex plus Ole-treated group, and vehicle group. Sections of thymus were taken 16 hours after dexamethasone injection and studied for histopathological and immunohistochemistry assessment. Result: Further characteristics of degeneration in thymocytes were observed in the Dex group compared with the Dex plus Ole group. Compared with the Dex group (10.94±3.35), positive staining for Bax in thymocytes decreased in Dex plus Ole group (2.64±1.26), but remained higher than the Ole (0.65±0.30) and vehicle (0.67±0.29) groups. Compared with the Dex group (2.94±0.42), positive staining for Bcl-2 in thymocytes increased in Dex plus Ole group (12.24±1.84) yet was lower than the Ole (14.94±1.54) and vehicle (18.93±3.54) groups. Conclusion: Our results suggest that dexamethasone-induced apoptosis is subsided by oleuropein.
Alireza Azizzadeh Delshad; Mohammad Hossein Ghaini; Marjan Heshmati
Volume 9, Issue 2 , April 2014, , Pages 124-132
Abstract
Background and Objective: The management of apoptotic cell death has been considered as a putative therapeutic strategy for cancer treatment. In the present study we investigated the putative pro-apoptotic effect of allicin, the main garlic organosulfur component with repeatedly claimed chemopreventive ...
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Background and Objective: The management of apoptotic cell death has been considered as a putative therapeutic strategy for cancer treatment. In the present study we investigated the putative pro-apoptotic effect of allicin, the main garlic organosulfur component with repeatedly claimed chemopreventive potency, on the human adenocarcinoma cell line HT29 as an apoptosis resistant cell line, in vitro.
Materials and Methods: The HT29 cells were incubated with different concentrations of allicin (0-40µg/ml) and for different time periods (6-48h) to investigate its effect on cell proliferation and apoptotic cell death.
Results: Five and 10µg/ml allicin could induce a significant cell death only after 12h, whereas concentrations of 20 and 40µg/ml resulted in a significant cell loss as soon as 6h. The results of the TUNEL assay, presented as percentage of apoptotic cells to total cell loss, indicated that concentrations ≥5µg/ml significantly increased the apoptotic features in time periods 6-24h, but after 48h no significant changes could be detected. The ratio of the sum of the apoptotic features of the four studied time points to the total cell loss calculated after 48h was about 0.5.
Conclusion: Allicin can induce apoptosis in a concentration- and time-dependent manner with most considerable effects achieved at 24h and by concentrations higher than 10µg/ml.
Majid Asadi-Shekaari; Hassan Eftekhar Vaghefi; Masoud Ezzat Abadi pour; Vahid Sheibani; Ali Shams Ara; Parisima Behbahani
Volume 6, Issue 4 , September 2011, , Pages 187-192
Abstract
Background and Objective: As one of the widely used drugs, aspirin (acetyl-salicylic acid, ASA) plays an important role in stroke treatment and prevention. In a previous study, we demonstrated ASA injection at 30 min after ischemia onset is neuroprotective. To determine whether the neurons protected ...
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Background and Objective: As one of the widely used drugs, aspirin (acetyl-salicylic acid, ASA) plays an important role in stroke treatment and prevention. In a previous study, we demonstrated ASA injection at 30 min after ischemia onset is neuroprotective. To determine whether the neurons protected by ASA had a normal ultrastructure, hippocampal CA1 pyramidal neurons were examined by Transmission Electron Microscope (TEM).
Material and Methods: Adult male wistar rats were divided into three different groups (6 animals/group): Sham-operated, control (48 MCAO+vehicle) and aspirin (48 MCAO + ASA). ASA (30 mg/kg) was injected 30 min after ischemia onset. The animals were killed 2 days after ischemia induction and their brain removed, processed, and examined under a TEM.
Results: Apoptotic changes were observed in rats not treated with ASA. In contrast, pyramidal neuron ultrastructure appeared normal in rats that exhibited neuroprotection (defined at the light microscope level) by ASA when studied two days after ischemia.
Conclusion: We conclude that administration of ASA after permanent focal cerebral ischemia remains a considerable therapeutic strategy.
Ghasem Azimi; Hesam Amini; Niosha Andalibi
Volume 5, Issue 1 , January 2010, , Pages 18-21
Abstract
Background and Objectives: Apoptosis of eosinophils is of significant value in assessing the airway inflammation in patients with asthma. Our purpose was to investigate the degree of expression of the Bcl-2 protein in sputum eosinophils during acute asthma exacerbation and its relationship with ...
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Background and Objectives: Apoptosis of eosinophils is of significant value in assessing the airway inflammation in patients with asthma. Our purpose was to investigate the degree of expression of the Bcl-2 protein in sputum eosinophils during acute asthma exacerbation and its relationship with exacerbation severity. Materials and Methods: The study was carried out in Mostafa Khomeini Hospital, Tehran, Iran in March 2008. Sputum was obtained from 15 asthmatic patients and 13 healthy subjects as a control group. Number of eosinophils was counted and Bcl-2+ eosinophils were stained using immunocytochemistry. Results: Sputum eosinophils and Bcl-2+ eosinophils were significantly higher in patients with acute exacerbation than controls (P<0.05). Conclusion: Bcl-2 prolongs survival and decreases apoptosis of airway eosinophils during acute asthma exacerbation. Eosinophil apoptosis and inhibition of Bcl-2 represent a target for new and effective therapeutic strategies of asthma.
Alireza Azizzadeh Delshad; Marjan Heshmati; Taki Tiraihi
Volume 3, Issue 2 , March 2008, , Pages 67-74
Abstract
Background and Objective: As apoptotic cell death is extremely involved in physiological development and many pathological situations such as cancer and neurodegenerative diseases, the understanding of its molecular machinery can be useful in designing new therapeutic strategies. The present ...
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Background and Objective: As apoptotic cell death is extremely involved in physiological development and many pathological situations such as cancer and neurodegenerative diseases, the understanding of its molecular machinery can be useful in designing new therapeutic strategies. The present study investigated the temporal expression of the proapoptotic protein Bax in adult spinal motoneurons. Materials and Methods: Following unilateral mid-thigh sciatic transection in adult rats, the incidence and nature of spinal motoneuron loss were evaluated by means of light microscopic cell count and electron microscopy 1 day, 1 week, 1 month and 3 months post-operatively. In all groups the temporal expression of Bax was immunohistochemically determined and the findings were compared with the results of the cell count. Results: Following axotomy the related motoneurons underwent chromatolytic changes which increased up to one month and diminished in the 3-month group. One day following axotomy the number of motoneurons did not show any significant reduction, but thereafter a progressive cell loss occurred, which was most prominent after three months. Electron microscopic study confirmed the ultrastructural apoptotic nature of cell death. Bax immunohistochemistry indicated an increasing immunoreactivity up to one month post-axotomy, but in 3-month group it was clearly diminished. Conclusion: Following transection of a peripheral nerve in adult animals, related motoneurons undergo chromatolytic changes which in some neurons may proceed to apoptotic cell death. Although the proapoptotic protein Bax has long been believed as the main apoptotic factor, other Bax-independent pathways may also participate in the axotomy-induced neuronal apoptosis which must not be ignored.
