Origanum vulgare extract induces apoptosis in Molt-4 leukemic cell line

OBJECTIVE: The purpose of this paper is to investigate the effect of Origanum vulgare extract as a plant with high anti-oxidant components on the induction of cell death in the Molt-4 cell line. BACKGROUND: Acute lymphocytic leukemia is characterized by the accumulation of a large number of lymphoblastic cells with high oxidant levels. METHODS: MTT assay was performed to determine the effect of O.vulgare extract on Molt-4 cells viability and the amount of 50%inhibitory concentration (IC50) was calculated. Changes in the expression of BAX and BCL-2 genes as involved in apoptosis and Nrf2 gene as a transcription factor of anti-oxidant genes in O.vulgare extract-treated Molt-4 cells were measured with Real-Time PCR. Treated Molt-4 cells were used to determine the stages of early and late apoptosis, and necrosis using acridine orange/ethidium bromide double staining. RESULTS: The results suggest survival inhibition and induction of apoptosis in Molt-4 cells treated with O.vulgare extract. Against Bax and Nrf2 genes expression, the expression of Bcl-2 gene has been reduced in Molt-4 cells following1/5 IC50 concentration of O. vulgare extract treatment. CONCLUSION: Given the oxidant drugs used in ALL treatment, and increased levels of oxidative stress in leukemic cells, induction of apoptosis by an anti-oxidant plant extract seems to be a promising way in leukemia treatment.

Simultaneous Exposure of Cultured Human Lymphoblastic Cells to Simulated Microgravity and Radiation Increases Chromosome Aberrations

During space travel, humans are continuously exposed to two major environmental stresses, microgravity (μG) and space radiation. One of the fundamental questions is whether the two stressors are interactive. For over half a century, many studies were carried out in space, as well as using devices that simulated μG on the ground to investigate gravity effects on cells and organisms, and we have gained insights into how living organisms respond to μG. However, our knowledge on how to assess and manage human health risks in long-term mission to the Moon or Mars is drastically limited. For example, little information is available on how cells respond to simultaneous exposure to space radiation and μG. In this study, we analyzed the frequencies of chromosome aberrations (CA) in cultured human lymphoblastic TK6 cells exposed to X-ray or carbon ion under the simulated μG conditions. A higher frequency of both simple and complex types of CA were observed in cells exposed to radiation and μG simultaneously compared to CA frequency in cells exposed to radiation only. Our study shows that the dose response data on space radiation obtained at the 1G condition could lead to the underestimation of astronauts’ potential risk for health deterioration, including cancer. This study also emphasizes the importance of obtaining data on the molecular and cellular responses to irradiation under μG conditions.

Photoluminescent Detection of Human T-Lymphoblastic Cells by ZnO Nanorods

The precise detection of cancer cells currently remains a global challenge. One-dimensional (1D) semiconductor nanostructures (e.g., ZnO nanorods) have attracted attention due to their potential use in cancer biosensors. In the current study, it was demonstrated that the possibility of a photoluminescent detection of human leukemic T-cells by using a zinc oxide nanorods (ZnO NRs) platform. Monoclonal antibodies (MABs) anti-CD5 against a cluster of differentiation (CD) proteins on the pathologic cell surface have been used as a bioselective layer on the ZnO surface. The optimal concentration of the protein anti-CD5 to form an effective bioselective layer on the ZnO NRs surface was selected. The novel biosensing platforms based on glass/ZnO NRs/anti-CD5 were tested towards the human T-lymphoblast cell line MOLT-4 derived from patients with acute lymphoblastic leukemia. The control tests towards MOLT-4 cells were performed by using the glass/ZnO NRs/anti-IgG2a system as a negative control. It was shown that the photoluminescence signal of the glass/ZnO NRs/anti-CD5 system increased after adsorption of T-lymphoblast MOLT-4 cells on the biosensor surface. The increase in the ZnO NRs photoluminescence intensity correlated with the number of CD5-positive MOLT-4 cells in the investigated population (controlled by using flow cytometry). Perspectives of the developed ZnO platforms as an efficient cancer cell biosensor were discussed.

Use of Four-Color Flow Cytometric Assay for Discrimination of Hematogone from Lymphoblast: Critical Issue for MRD Assessment in B-ALL Patients

Background: Hematogones are normal B-cell precursor which can be seen in different physiological and pathological conditions. Due to variation in B-cell acute lymphoblastic leukemia (B-ALL) blasts immunophenotyping and interference of hematogones in minimal residual disease (MRD) assessment, precise discrimination of hematogones is very crucial. The purpose of this study was to evaluate the expression pattern of surface markers in hematogones and compare them with lymphoblasts. Material and Methods: In this applied study, flow cytometric analysis was performed using Coulter FC-500 and MXP software in 4-color combination and 6 different tubes. In this study, 85 patients diagnosed with acute lymphoblastic leukemia were evaluated. Out of these patients, 45 were boys and 40 were girls. Patients aged from 1 to 15 years old. In addition, 27 bone marrow samples from other patients aged 4 to 18 years were included in this investigation. These samples had been obtained for other diagnostic purposes, such as immune thrombocytopenic purpura and juvenile idiopathic arthritis. Results: During flow cytometric analysis, hematogones showed expressions of CD19, CD20, CD22, CD10, CD45, CD81, CD123, CD9, CD34 (partial expression), and tdt (partial expression). In these patients, hematgones were negative for CD66c expression. Lymphoblastic cells were positive for CD19, CD20 (in some cases), CD22, CD10, CD45, CD81, CD123, CD58, CD9, CD66c, CD34 (in most cases), and TDT. CD81 mean fluorescence intensity (MFI) in hematogones was higher than that in lymphoblasts. (112.5 (30-251) vs. 17.5 (5-30); P<0.0001) Conclusion: According to findings of this study, it seems that the use of CD81, CD58, CD123, CD66c, CD9, and CD81 MFI in combination with B-Cells associated markers can be very effective in differentiating hematogone from lymphoblast.

A Case of Lymphocytic Leukemia in a Bearded Dragon (Pogona vitticeps) and a Review of Literature

The present paper reports the case of a 3 years old, female Bearded Dragon (Pogona vitticeps) presenting lethargy, anorexia, weight loss, and anemia and marked leukocytosis at CBC. The majority of leukocytes were lymphocytic/lymphoblastic cells (97%). Immunocytochemical staining of blood smears marked for CD3 (neg) and CD79a (pos) suggested immunophenotype B. The patient died after one month from diagnosis. Histology evidenced lymphoid infiltration in the heart, spleen, liver, kidneys and gut. In addition, in the bone marrow a massive infiltration of lymphoid cells confirmed the diagnosis of leukemia. Immunohistochemistry confirmed the CD79a positivity of a large part of infiltrating lymphoid cells indicating a B cells immunophenotype of the neoplastic population. The presence of lymphocytosis and multiorgan infiltration supported the diagnosis of lymphocytic leukemia. Finally, a revision of the literature has also been made.