Combinatorial targeting of telomerase and DNA-PK induces synergistic apoptotic effects against Pre-B acute lymphoblastic leukemia cells.

BACKGROUND: Due to the high demand for novel approaches for leukemia-targeted therapy, this study investigates the impact of DNA-PK inhibitor NU7441 on the sensitivity of pre-B ALL cells to the telomerase inhibitor MST-312. METHODS: The study involved NALM-6 cells treated with MST-312 and NU7441, assessing their viability and metabolic activity using trypan blue and MTT assays. The study also evaluated apoptosis, gene expression changes, and DNA damage using flow cytometry, qRT-PCR, and micronucleus assays. The binding energy of MST-312 in the active site of telomerase was calculated using molecular docking. RESULTS: The study's findings revealed a synergistic decline in both cell viability and metabolic activity in NALM-6 cells when exposed to the combined treatment of MST-312 and NU7441, and this decrease occurred without any adverse effects on healthy PBMC cells. Furthermore, the combination treatment exhibited a significantly higher induction of apoptosis than treatment with MST-312 alone, as observed through flow cytometry assay. qRT-PCR analysis revealed that this enhanced apoptosis was associated with a notable downregulation of Bcl-2 expression and an upregulation of Bax gene expression. Moreover, the combination therapy decreased expression levels of hTERT and c-Myc genes. The micronucleus assay indicated that the combination treatment increased DNA damage in NALM-6 cells. Also, a good conformation between MST-312 and the active site of telomerase was revealed by docking data. CONCLUSIONS: The study suggests that simultaneous inhibition of telomerase and DNA-PK in pre-B ALL presents a novel targeted therapy approach.

The Serum Level of CXCL9, CXCL10, and CXCL11 and the Expression of CXCR3 of Peripheral Blood Mononuclear Cells in Brucellosis Patients.

Brucella spp. can replicate in human endothelial cells, inducing an inflammatory response with increased expression of chemokines. Although Brucella infects humans, its ability to induce the production of chemokines by lung cells is unknown. Therefore, the current investigation was designed to examine the association between brucellosis and CXCL9, 10, and 11 chemokines. The patient group included 71 patients suffering from Brucella infection and the control group consisted of 50 healthy ranchers from the same geographical area. Serum levels of CXCL9, CXCL10, and CXCL11 were analyzed by ELISA. The fold changes of CXCR3 expression against ß-actin were determined by real-time-PCR technique. Western blotting analysis was also applied for evaluating the expression of CXCR3 at protein level. The results of this study showed that the serum levels of CXCL9, CXCL10, and CXCL11 are significantly increased in acute brucellosis patients in comparison to control as indicated by ELISA test, mRNA levels of CXCR3 by Real-time PCR as well as protein levels of CXCR3 by Western blot analysis. According to findings, these chemokines have the potential to serve as markers for brucellosis patients. Taken together, cytokine/chemokine network was active in acute brucellosis patients, and it is suggested to evaluate other cytokines in future studies.

Radioprotective Effect of Zinc Nanoparticles on Ionizing Radiation-induced Nephrotoxicity in Mice.

INTRODUCTION: Ionizing radiation (IR) causes oxidative stress in kidneys and subsequently disrupts renal function. The use of green synthesized zinc nanoparticles (Zn NPs) with antioxidant properties may reduce the damage caused by IR. METHODS: Thirty-six mice were kept in a standard situation and divided into 6 groups: 1: Control; 2-4: receiving 5 mg/kg, 10 mg/kg, and 25 mg/kg of Zn NPs with IR; 5: receiving 5 mg/kg of ZnSO4 with IR; and 6: IR. After 15 days, half of the animals in each group were sacrificed and their blood samples isolated to evaluate the plasma urea and creatinine levels. The kidneys were kept for evaluating the glutathione (GSH), malondialdehyde (MDA), superoxide dismutase (SOD), and catalase (CAT) levels; on 21st day, the rest of the animals were sacrificed and their kidneys removed for histological assessments. RESULTS: IR decreased GSH content, increased MDA level, and reduced SOD and CAT activity. On the other hand, Zn NPs at 10 and 25 mg/kg doses increased GSH, decreased MDA, and enhanced SOD and CAT activities. Zn NPs treatment at 10 and 25 mg/kg doses decreased the plasma urea and creatinine levels induced by IR. Moreover, Zn NPs significantly decreased the level of urea and creatinine in irradiated mice in comparison with IR alone (p < 0.05). The main histopathological results were tubular and glomerular atrophy and interstitial fibrosis in irradiated mice, while tubular degeneration and atrophy were less frequent in Zn NPs + IR group than in IR group alone. CONCLUSION: Zn NPs treatment, especially at 25 mg/kg dose, attenuates the side effect of IR on kidneys through reducing oxidative stress factors, biochemical, and histopathological changes.

Radioprotective effect of a novel and green bio-nanohybrid, chitosan/silver/cobalt complex, based on Ferulago angulate plant.

A novel bio-nanocomposite was developed by incorporating the extracted nanochitosan from shrimp wastes with Schiff base cobalt complex (Chit-Co complex). The phytosynthesis of Chit-Co complex/Ag bio-nanocomposite was designed utilizing Chit-Co complex at the presence of Ferulago angulate extraction and characterized by AFM, SEM, EDAX, TEM, FT-IR, and elemental analysis. The radioprotective application of this bio-nanocomposite on human lymphocyte cells was evaluated using micronucleus (MN) assay. Total antioxidant activities of it were evaluated using FRAP and DPPH assays. Chit-Co complex/Ag bio-nanocomposite significantly decreased the frequency of micronuclei in human lymphocytes exposed to ionization irradiation (IR). The highest protection was observed at 200 µg/ml. Also, maximum antioxidant activities of bio-nanocomposite were provided at the same dose. These data exhibit the radioprotective effect of a bio-nanocomposite based on wastes of living organisms can be an excellent radioprotective agent, which can protect the normal cells of human against the genetic damage by IR.

Inhibition of DNA damage response pathway using combination of DDR pathway inhibitors and radiation in treatment of acute lymphoblastic leukemia cells.

An alarming increase in acute lymphoblastic leukemia cases among children and adults has attracted the attention of researchers to discover new therapeutic strategies with a better prognosis. In cancer cells, the DNA damage response (DDR) pathway elements have been recognized to protect tumor cells from various stresses and cause tumor progression; targeting these DDR members is an attractive strategy for treatment of cancers. The inhibition of the DDR pathway in cancer cells for the treatment of cancers has recently been introduced. Hence, effective treatment strategies are needed for this purpose. Chemotherapy in combination with radiotherapy is considered a potential therapeutic strategy for acute leukemia. This review aims to assess the synergistic effects of these inhibitors with irradiation for the treatment of leukemia.

Lay abstract Acute lymphoblastic leukemia (ALL) is a blood malignancy that is caused by the high proliferation of lymphoid precursors in bone marrow and peripheral blood, and it is one of the common malignancies among children and adults. Novel therapeutic strategies are increasingly being studied to suppress cancer cells. DNA damage response pathways have developed in cancer cells that protect tumor cells from chemotherapy and radiotherapy and cause relapse and treatment failure. Effective drugs in ALL treatment can suppress cancer cells through several mechanisms, such as inducing DNA damage in cancer cells. However, due to the active repair systems in cancer cells, treatment may fail. Recently, the inhibition of these repair pathways and the combination of DNA damage response pathway inhibitors with chemotherapy and radiotherapy in cancer treatment have been studied. The purpose of this review is to outline the synergistic effects of DNA damage response pathway inhibitors and radiation in the death of leukemic cells.

Comparative assessment of a 99mTc labeled H1299.2-HYNIC peptide bearing two different co-ligands for tumor-targeted imaging.

Peptides are a class of targeting agents that bind to cancer-specific cell surfaces. Since they specifically target cancer cells, they could be used as molecular imaging tools. In this study, the 15-mer peptide Ac-H1299.2 (YAAWPASGAWTGTAP) was conjugated with HYNIC via lysine amino acid on C-terminus and labeled with 99mTc using tricine and EDDA/tricine as the co-ligands. These radiotracers were evaluated for potential utilization in diagnostic imaging of ovarian cancer cells (SKOV-3). The cell-specificity of these radiolabeled peptides was determined based on their binding on an ovarian cancer cell line (SKOV-3), and displaying a low affinity for lung adenocarcinoma cell line (A549) and breast cancer cell line (MCF7). Biodistribution studies were conducted in normal mice as well as in nude mice bearing SKOV-3 ovarian cancer xenografts. HYNIC-peptide was labeled with 99mTc with more than 99% efficiency and showed high stability in buffer and serum. We observed nanomolar binding affinities for both radiolabeled peptides. The tumor uptakes were 3.27%±0.46% and 1.55%±0.20% for tricine and 2.34±1.1% and 1.09%±0.18% for EDDA/tricine at 1 and 4h after injection, respectively. A higher tumor to background ratio and lower radioactivity in the blood were observed for EDDA/tricine co-ligands, leading to clear tumor visualization in imaging with injection of this peptide. This new 99mTc-labeled peptide selectively targeted ovarian cancer and introduction of a (EDDA/tricine) as a co-ligand improved the pharmacokinetics of 99mTc-labeled H1299.2 for tumor imaging in animals.

Pistacia and its Combination with Cisplatin: A Potential Anticancer Candidate by Modulating Apoptotic Genes.

INTRODUCTION: Many bioactive phytochemicals have essential significance for handling various diseases and developing new drugs. The aim was to investigate the anti-tumor activity and the underlying mechanisms of pistachio pericarp extract (PPE) and pistachio kernel extract (PKE) alone and combined with cisplatin (CP) in the treatment of prostate cancer. METHODS: The effects of the PPE, PKE, and CP alone and PPE and PKE in combination with CP (PPE+CP and PKE+CP) on the proliferation of PC-3 cells were determined using the MTT assay. The fold changes of BAX, BCL-2, P53, KLK2, TNF, TGF, and NANOG expression against ß-actin were determined by real-time technique. Data were analyzed by two-way ANOVA and repeated measure tests. RESULTS: These research results indicated that a greater anti-proliferative effect of the PPE and PKE was shown in combination with CP compared with treatments using the PPE and PKE or CP alone. The extracts and Cisplatin in vitro had good synergistic effects on the inhibition of the proliferation of PC-3 cells. The IC50 values of PKE+CP were 4.141, 2.140, and 0.884 ug/mL, and PPE+CP were 2.754, 2.061, and 0.753 ug/mL after 24h, 48 h, and 72h treatment, respectively. Also, this result presented that the mRNA expression of BAX and P53 increased, and BCL-2, KLK2, TNF, TGF, and NANOG decreased in PC-3 cells. CONCLUSIONS: The finding of this research showed for the first time the anti-carcinogenesis effects of separately and in the combination of PPE, PKE, and CP on the PC-3 prostate cancer cells via modulating some genes and that it may be nominated for the herbal anti-cancer medications.

The Glucose-Regulated Protein78 (GRP78) in the Unfolded Protein Response (UPR) Pathway: A Potential Therapeutic Target for Breast Cancer.

Amongst all types of cancers, breast cancer is recognized as the most common cancer and a principal cause of morbidity and mortality in women. Endoplasmic reticulum (ER) stress pathways are primarily activated in cancer cells and activate a signaling network called the unfolded protein response (UPR). Many tumors, by activating the UPR pathway, allow them to adapt and grow under stressful conditions. UPR is usually inactive in non-tumor cells, while it is active in tumor cells, so it is appropriate to develop new breast cancer therapies. A protein that regulates UPR is 78 KDa Glucose-Regulated Protein (GRP78). Usually, the GRP78 level in the cell is relatively low but increases significantly under stresses that affect the ER and calcium homeostasis, and increases resistance to chemotherapy. GRP78 drug suppressors could provide promising anticancer therapeutics. Therefore, understanding the molecular mechanism of GRP78 in cancer and identifying drugs that target GRP78 is essential for the treatment of breast cancer. In this review, we investigate the role of GRP78 in the pathogenesis of breast cancer.

Indole-3-carbinol (I3C) as Leukemia Therapeutic Agents: Review.

Leukemia or blood cancer was initially discovered in 1845 and this malignancy was reported in patients who had an amplified number of blood cells, in particular, White Blood Cells (WBC), due to this disease. The event of leukemia was further identified as a malignant hematopoietic disorder due to both uncontrolled and unlimited proliferation in combination with a lack of differentiation of the leukemic stem cells. Furthermore, 75 to 80% of the global population use herbal remedies as primary therapy, mainly because of their better efficiency and satisfaction, which elevate the human body symmetry with the minimum unwanted adverse effects. For the control of cancer, plant products, and fruits have been considered promising tools and are being consumed for centuries. Several plant extracts are also being used for the therapy and prevention of different types of known cancers. Indole-3-carbinol (I3C) is a natural material obtained from Brassica diversity of vegetables and has been reported to serve as a promising cancer preventative agent. In the present review, the authors mainly tried to focus on and emphasize I3C applications in leukemia treatment.

The Role of Genetics and Synergistic Effect of Targeting Common Genetic Mutations in Acute Lymphoblastic Leukemia (ALL).

Increasing concern regarding non-treatment and relapse in Acute Lymphoblastic Leukemia (ALL) among children and adults has attracted the attention of researchers to investigate the genetic factors of ALL and discover new treatments with a better prognosis. Nevertheless, the survival rate in children is more than in adults; therefore, it is necessary to find new potential molecular targets with better therapeutic results. Genomic analysis has enabled the detection of different genetic defects that are serious for driving leukemogenesis. The study of genetic translocation provides a better understanding of the function of genes involved in disease progression. This paper presents an overview of the main genetic translocations and dysregulations in the signaling pathways of ALL. We also report the inhibitors of these main translocations and evaluate the synergistic effect of chemical inhibitors and gamma-ray irradiation on ALL.

Investigating Association Level of CXCL12 with its SDF-1α 3'A Genetic Variant and CXCL10 with its 1443 Promoter Polymorphism in Type-1 Diabetes.

BACKGROUND: Type-1 Diabetes Mellitus (T1DM) is an autoimmune and heterogeneous disorder. In the present study, we aimed to examine whether there exists an association between serum CXCL10 (IP-10) level and its promoter polymorphism at position-1443 along with CXCL12 and its known SDF-1 3' A genetic variant as an angiogenesis chemokine in T1DM patients. METHODS: Blood specimens were collected from 209 unrelated T1DM patients and 189 healthy subjects. The DNA samples were extracted from the subjects and analyzed for CXCL10 and CXCL12 polymorphisms using PCR-RLFP. The serum concentrations of CXCL10 and CXCL12 were also analyzed with ELISA. RESULTS: Following expert opinion and data analysis, we found significant differences between A/A, A/G, and G/G genotypes with A and G alleles of polymorphisms at position +801 (SDF-1α3'A) in CXCL12. No association was reported between CXCL10/-1443 promoter polymorphism and T1DM. In our assessment of promoter polymorphism, both T1DM patients and controls had GG genotypes in CXCL10/-1443. When patients were compared with controls, both serum CXCL10 and CXCL12 levels were found to be increased in type 1 diabetes with complications. Levels were not increased in patients without complications. CONCLUSION: Both CXCL10 and CXCL12 play fundamental roles in T1DM pathogenesis. Only the CXCL12 3'A (SDF-1α3'A) polymorphism is possibly necessary for the pathogenesis of T1DM, while the CXCL10-1443 promoter polymorphism is not.

Ferulago angulata as a Good Radioprotector Against Genotoxicity.

INTRODUCTION: Natural products can be used as radioprotector agents because of containing phenolic compounds and several flavonoids with antioxidant properties. When the normal cells are exposed to ionizing radiation, they generate free radicals and reactive oxygen species that can cause damage in DNA, which leads to cellular dysfunction or even cell death. However, it is necessary to identify new radioprotective agents to protect normal cells. Ferulago angulata (F. angulata), a medicinal plant, can be used as a new radioprotective agent. PURPOSE: In this study, the radioprotective effect of F. angulata was evaluated against genotoxicity and oxidative stress induced by ionizing radiation in human blood lymphocytes. METHODS: The antioxidant activity of F. angulata was assayed using FRAP and DPPH methods. Then, the human blood samples were incubated with F. angulata at different concentrations (25, 50, 100, and 200 µM) and subsequently exposed to IR at a dose of 2Gy. The radioprotective effect of F. angulata on the exposed cells was assessed by the micronucleus (MN) method. Also, biomarkers of oxidative stress in the exposed cells were evaluated by malondialdehyde (MDA) and superoxide dismutase (SOD) methods. RESULTS: Our findings showed that F. angulata reduced the frequency of MN induced by IR in exposed cells. At a 200 µM concentration of F. angulata, the maximum reduction in the frequency of MN (63.11%) was observed that demonstrated a high degree of radioprotection. Afterward, pretreatment at 200 µM concentration of F. angulata inhibited oxidative stress in irradiated lymphocytes, leading to a reduction in MN frequency and MDA levels while SOD activity was enhanced in the exposed cells. CONCLUSION: F. angulata as a natural radioprotective agent can protect normal cells against reactive oxygen species and genetic damage induced by IR.

Effect of Methadone Maintenance on Expression of BDNF and CREB Genes in Brain VTA of Male Morphine Treated Rats.

BACKGROUND: Morphine independently reduces the expression level of Brain-derived Neurotrophic Factor (BDNF) and Cyclic-AMP Response Element Binding protein (CREB). BDNF and CREB play a vital role in protecting and regulating the proper functioning of neurons. There has not been any study on the effect of methadone maintenance treatment and its comparison with morphine. Therefore, this study was conducted to examine the effect of methadone maintenance on the expression of BDNF and CREB genes in brain VTA of male morphine treated rats. METHODS: In this study, 24 Wistar rats (200-250g) were assigned to three experimental groups: 1) Animals without morphine treatment (control); 2) Morphine treated animals (10 mg/kg, twice/day through subcutaneous injection for 21 days); 3) Animals under methadone maintenance after treatment with morphine (maintenance dose of methadone was achieved during 14 days equal to 1 mg per 100 ml at the first week and 2.5 mg per 100 ml at second week). To evaluate the expression of BDNF and CREB genes, real time PCR method was used, and ELISA was applied to measure the serum level of BDNF protein at the end of the experiment. RESULTS: According to the findings of this study, similar to morphine treated group, methadone maintenance in morphine treated animals led to a significant reduction in the expression of BDNF and CREB genes at VTA as well BDNF serum level compared with the control group. CONCLUSION: It was concluded that methadone, like morphine, causes a significant reduction in the expression of BDNF and CREB genes in the brain VTA area of rats as well as BDNF serum level compared with the control group.

The first and low cost copper Schiff base/manganese oxide bio nanocomposite from unwanted plants as a robust industrial catalyst.

In this paper, the first bio-nano colloid including manganese oxide nanoparticles and copper nanocomplex in the presence of Amaranthus spinosus as one unwanted plant was prepared (Mn3O4/CuL bio-nanocolloid). As-prepared bio nanocolloid was analysed completely by different techniques such as FT-IR, ICP-AES, SEM, EDX, TEM and elemental analysis to have the size, structure, morphology and elements in this compound. This bio-nanocolloid showed high catalytic activity towards green oxidation reactions of alcohols using hydrogen peroxide under solvent free conditions. The longevity, easy and practical recoverability of the solid catalyst was also confirmed for six times. The natural starting material for a long-term stability catalyst preparation, using ideal oxidant (H2O2), solventless and easy work up show the great potential in scalability for actual industries applications.

Radioprotective Potential of Sulindac Sulfide to Prevent DNA Damage Due to Ionizing Radiation.

Introduction: The ionizing radiation exposure of the normal cell causes damage to DNA, which leads to cell dysfunction or even cell death. However, it is necessary to identify new radio protectives in order to protect normal cells. Sulindac sulfide (SS) is a metabolite of sulindac (a non-steroidal anti-inflammatory drug) known as a cyclooxygenase inhibitor. Free radicals and reactive oxygen species are generated in the IR-exposed cells. Also, the induced inflammation process causes damage in DNA. PURPOSE: In this research, the radioprotective effect of SS was investigated against genotoxicity and lipid peroxidation induced by ionizing radiation in the human blood lymphocytes. METHODS: In this study, the human blood samples were pretreated with SS at different concentrations (10, 25, 50, 100 and 250 µM) and then were exposed to IR at a dose of 1.5 Gy. The micronucleus (MN) assay was used to indicate the radioprotective effects of SS on exposed cells. Total antioxidant activity of the SS was measured by using FRAP and DPPH assay. Also, the malondialdehyde (MDA) levels and the activity of superoxide dismutase (SOD) on the exposed cells were evaluated. RESULTS: It was found that SS decreased the percentage of MN induced by IR in exposed cells. Maximum reduction in the frequency of MN was observed at 250 µM of SS (87%) that provides the highest degree of protection against IR. On the other hand, pretreatment at 250 µM of SS inhibited IR-induced oxidative stress, which led to a decrease in the MN frequencies and MDA levels, while SOD activity showed an increase in the exposed cells. CONCLUSION: It could be concluded that SS as a good radioprotective agent protects the human normal cells against the oxidative stress and genetic damage induced by IR.

The Influence of Co-Ligands on Improving Tumor Targeting of 99mTc-HYNIC Conjugated Peptides.

Technetium-99m (99Tc) is widely used as a radionuclide for labeling of biomolecules in nuclear medicine practice. 6-hydrazinopyridine-3-carboxylic acid namely HYNIC is one of the best bifunctional chelating agents for attachment of 99mTc to the biomolecules such as peptides and proteins. Since HYNIC can only occupy one or two coordination sites of 99mTc, co-ligands are needed to complete the coordination sphere of the technetium. Selection of co-ligands can be impressive on the stability, lipophilicity and biodistribution of 99mTc-HYNIC conjugated peptides. The aim of this review was to explain the chemical and biological effects of various co-ligands on characteristics of 99mTc-HYNIC conjugates for tumor or diseases imaging.