Modulation of wheatgrass (Triticum aestivum Linn) toxicity against breast cancer cell lines by simulated microgravity.

This study scrutinizes the effects of simulated microgravity on the antioxidant and cytotoxic potential, along with the phytochemical content of wheatgrass (Triticum aestivum Linn). To imitate microgravity, wheatgrass seeds were germinated in a 3D-clinostat at different rotations per minute (5, 10, 15, and 20 rpm), together with terrestrial gravity control, over 10 days. After germination, the methanolic extracts were analyzed using UPLC-Triple Quad LCMS for their phytochemical composition and tested for their hydrogen peroxide, nitric oxide, and DPPH scavenging activities. The cytotoxic effects of these extracts were evaluated against normal skin fibroblasts, normal breast cells (MCF-10), and breast cancer cells (MCF-7 and MDA-231). The findings showed an extended root growth in wheatgrass germinated under microgravity (WGM) compared to under gravity (WGG). Additionally, WGM extracts demonstrated increased H2O2-, NO-, and DPPH-scavenging activities and a higher content of polyphenols and flavonoids than WGG extracts. These effects were amplified with an increase in clinostat rotations. Moreover, WGM extracts were found to contain a unique set of bioactive compounds (compounds that were detected in the microgravity-germinated wheatgrass but were either absent or present in lower concentrations in wheatgrass germinated under standard gravity conditions.), including pyridoxine, apigenin, and tocopherol, among others, which were absent in WGG. The UPLC-Triple Quad LCMS analysis revealed these unique bioactive compounds in WGM. Notably, WGM extracts showed enhanced cytotoxic effects against normal skin fibroblasts, normal MCF-10, MCF-7, and breast cancer MDA-231 cell lines, with increased cytotoxicity correlating with the number of clinostat rotations. Particularly, WGM extract (at 20 rpm) demonstrated significantly stronger cytotoxicity against MCF-7 breast cancer cells. Further in-depth gene expression analysis of MCF-7 cells exposed to WGM revealed a significant downregulation of genes integral to breast cancer pathways, tyrosine kinase signaling, and DNA repair, complemented by upregulation of certain cell survival and cytotoxic genes. These alterations in genetic pathways associated with cell survival, hormone responses, and cancer progression may elucidate the enhanced cytotoxicity observed in WGM extracts. Our findings underscore the potential of microgravity as a tool to enhance the cytotoxic capabilities of wheatgrass against cancer cell lines, presenting a promising direction for future research in the field of space biology and its implications for terrestrial health.

Correlates of SARS-CoV-2 Variants on Deaths, Case Incidence and Case Fatality Ratio among the Continents for the Period of 1 December 2020 to 15 March 2021.

The outbreak of coronavirus disease 2019 (COVID-19), by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has quickly developed into a worldwide pandemic. Mutations in the SARS-CoV-2 genome may affect various aspects of the disease including fatality ratio. In this study, 553,518 SARS-CoV-2 genome sequences isolated from patients from continents for the period 1 December 2020 to 15 March 2021 were comprehensively analyzed and a total of 82 mutations were identified concerning the reference sequence. In addition, associations between the mutations and the case fatality ratio (CFR), cases per million and deaths per million, were examined. The mutations having the highest frequencies among different continents were Spike_D614G and NSP12_P323L. Among the identified mutations, NSP2_T153M, NSP14_I42V and Spike_L18F mutations showed a positive correlation to CFR. While the NSP13_Y541C, NSP3_T73I and NSP3_Q180H mutations demonstrated a negative correlation to CFR. The Spike_D614G and NSP12_P323L mutations showed a positive correlation to deaths per million. The NSP3_T1198K, NS8_L84S and NSP12_A97V mutations showed a significant negative correlation to deaths per million. The NSP12_P323L and Spike_D614G mutations showed a positive correlation to the number of cases per million. In contrast, NS8_L84S and NSP12_A97V mutations showed a negative correlation to the number of cases per million. In addition, among the identified clades, none showed a significant correlation to CFR. The G, GR, GV, S clades showed a significant positive correlation to deaths per million. The GR and S clades showed a positive correlation to number of cases per million. The clades having the highest frequencies among continents were G, followed by GH and GR. These findings should be taken into consideration during epidemiological surveys of the virus and vaccine development.

Continuous Hypoxia and Glucose Metabolism: The Effects on Gene Expression in Mcf7 Breast Cancer Cell Line.

BACKGROUND: Hypoxia (deprived oxygen in tissues) may induce molecular and genetic changes in cancer cells. OBJECTIVE: To Investigate the genetic changes of glucose metabolism in breast cancer cell line (MCF7) after exposure to continuous hypoxia (10 and 20 cycles exposure of 72 hours continuously on a weekly basis). METHODS: Gene expression of MCF7 cells was evaluated using real-time polymerase chain reactionarray method. Furthermore, cell migration and wound healing assays were also applied. RESULTS: It was found that 10 episodes of continuous hypoxia activated the Warburg effect in MCF7 cells, via the significant up-regulation of genes involved in glycolysis (ANOVA, p value < 0.05). The molecular changes were associated with the ability of MCF7 cells to divide and migrate. Interestingly, after 20 episodes of continuous hypoxia, the expression glycolysis mediated genes dropped significantly (from 30 to 9 folds). This could be attributed to the adaptive ability of cancer cells. CONCLUSION: It is concluded that 10 hypoxic episodes increased the survival rate and aggressiveness of MCF7 cells and induced the Warburg effect by the up-regulation of the glycolysis mediating gene expression.

AOX1 and XDH Enzymes Genotyping and its Effect on Clinical Response to Azathioprine in Inflammatory Bowel Disease Patients Among Jordanian Population.

BACKGROUND AND OBJECTIVES: Inflammatory bowel disease (IBD) is a set of chronic inflammatory gastrointestinal disorders, which include ulcerative colitis (UC) and Crohn's disease (CD) that affects many patients worldwide with a peak incidence in early adult life. The immunosuppressant drug Azathioprine (AZA) represents one of the most useful drugs in the management of IBD. It is metabolized by many enzymes like AOX1, and XDH enzymes, the variation in the metabolism of AZA may contribute to inter-individual variation in response to this treatment. This study aims to find out if there is an association between certain AOX1 and XDH polymorphisms and AZA response in Jordanian IBD patients. METHODS: One hundred IBD patients aged between (17-72) years and taking AZA were enrolled and genotyped for AOX13404G, XDH1936C and XDH2107C polymorphisms using DNA Sequencing (Sanger) method. RESULTS AND CONCLUSION: This study revealed that 16% of our patients were non-responders to AZA; they needed an alternative therapy (biological agent) or steroids along with AZA. There was no statistically significant association (p-value>0.05) between the AOX1 3404G, XDH 1936C and XDH 2107C polymorphisms and the response to AZA among Jordanian IBD patients. Finally, the study showed an association between the age of the patient and the response to AZA (p-value=0.013).

Effect of simulated microgravity on the antidiabetic properties of wheatgrass (Triticum aestivum) in streptozotocin-induced diabetic rats.

Microgravity affects plant growth and content. A three-dimensional clinostat was used at 4 rotations/min to rotate the seeds of Triticum aestivum cultivar (Ammon) in three dimensions for 7 days, following which the antioxidant activities of ethanolic extracts were evaluated using both nitric oxide- and hydrogen peroxide-scavenging activities. The antidiabetic activities of ethanolic extracts were evaluated by measuring the concentration of plasma glucose, insulin, C peptide, and glycated hemoglobin (HbA1c); determining the number of ß cells in the pancreatic islets; and performing the glucose tolerance test. Furthermore, the effects of the ethanolic extracts on the lipid profile and liver function were estimated. After rats were sacrificed, their pancreases were isolated and used for histopathological processing. The results indicated that the antioxidant potential and antioxidant metabolite content were significantly increased under microgravity conditions in comparison to those under normal gravity conditions. Rats treated with an extract of wheatgrass (T. aestivum) germinated over a period of 6-10 days under microgravity (WGM) showed a significant reduction in the levels of serum glucose, HbA1C, urea, creatinine, aspartate aminotransferase and alanine aminotransferase, and insulin resistance compared to rats treated with an extract of wheatgrass germinated under gravity. Additionally, the total cholesterol and low-density lipoprotein cholesterol levels were significantly decreased. In contrast, high-density lipoprotein cholesterol, C-peptide, and insulin levels rose significantly after treatment with T. aestivum germinated under microgravity. WGM is a promising potential diabetic treatment without side effects with a low manufacturing cost.

Effects of Intermittent Hypoxia on Expression of Glucose Metabolism Genes in MCF7 Breast Cancer Cell Line.

BACKGROUND: Hypoxic condition induces molecular alterations which affect the survival rate and chemo-resistant phenotype of cancer cells. OBJECTIVE: The aim of this study is to investigate the influence of intermittent hypoxic conditions on the expression of glucose metabolism genes in breast cancer MCF7 cell line. METHODS: The gene expression was analyzed using a polymerase chain reaction-array method. In addition, the cell resistance, survival and migration rates were examined to assure the hypoxic influence on the cells. RESULTS: 30 hypoxic episodes induced the Warburg effect through significant (p-value < 0.05) upregulation of the expression of PCK2, PHKG1, ALDOC, G6PC, GYS2, ALDOB, HK3, PKLR, PGK2, PDK2, ACO1 and H6PD genes that are involved in glycolysis, were obtained. Furthermore, the expression of the major gluconeogenesis enzyme genes was significantly (ANOVA, p-value < 0.05) downregulated. These molecular alterations were associated with increased MCF7 cell division and migration rate. However, molecular and phenotypic changes induced after 30 episodes were normalized in MCF7 cells exposed to 60 hypoxic episodes. CONCLUSION: It is concluded, from this study, that 30 intermitted hypoxic episodes increased the survival rate of MCF7 breast cancer cells and induced the Warburg effect through upregulation of the expression of genes involved in the glycolysis pathway. These results may increase our understanding of the molecular alterations of breast cancer cells under hypoxic conditions.

TPMT Genotype and Adverse Effects of Azathioprine among Jordanian Group.

BACKGROUND: Inflammatory Bowel Disease (IBD) is a common disease affecting many patients. This disease is treated by azathioprine and TPMT genetic polymorphism affecting the patient's tolerance. The aim of this study is to investigate the importance of TMPT genotyping in reducing the incidence of adverse effects of azathioprine. METHODS: One hundred and forty-one IBD patients were followed for azathioprine Adverse Drug Reaction (ADR). Patients were genotyped for TPMT*2, TPMT*3A, TPMT*3B, TPMT*3C. RESULTS: The frequency of Azathioprine adverse effect was about 35.5%. An association between TPMT genotypes 1/3A and 3B/3B and azathioprine related bone marrow suppression was found (P value ≤ 0.05). CONCLUSION: The findings suggest that there was a significant association between TPMT genotypes 1/3A and 3B/3B and azathioprine related bone marrow suppression.

The effect of green tea consumption on the expression of antioxidant- and inflammation-related genes induced by nicotine.

The aim of this study is to investigate the protective effect of green tea (GT) against the toxicity of nicotine. BALB/c mice were divided into four groups. Group I received food and water intake ad libidium, Group II received GT solution at a dose of 1 ml/kg body weight orally twice a day via gastric gavage, Group III was injected intraperitoneally with nicotine (2.5 mg/kg) once per day for 4 weeks, and Group IV received both nicotine and GT; GT was introduced using gastric gavage 1 hr before and 1 hr after the nicotine injection. The administration of nicotine altered the cellular antioxidant defense system by inducing inflammation and damage in the tissues of liver, lungs, and kidneys. In addition, nicotine treatment significantly enhanced the expression antioxidant- and inflammation-related genes. There were significant improvements when the nicotine-exposed mice treated with GT. PRACTICAL APPLICATIONS: In this study, it is revealed that the administration of nicotine altered the cellular antioxidant defense system by inducing inflammation manifested by the infiltration of inflammatory cells and damage seen in liver, lungs, and kidneys. GT contributed to the reduction of toxicity of nicotine, probably mediated by free radicals, through downregulation of nicotine-induced upregulated antioxidant- and inflammation-related genes. Never the less, further in depth investigation on characterization of the active constituents of GT responsible for their effect seen here and the mechanism that contributes to the effects seen in this reports is highly demanded. Furthermore, GT extract could be considered as a dietary supplement for the reduction of nicotine toxicity among cigarette smoker.

The effect of cycling hypoxia on MCF-7 cancer stem cells and the impact of their microenvironment on angiogenesis using human umbilical vein endothelial cells (HUVECs) as a model.

BACKGROUND: Breast cancer is the most common type of cancer among females. Hypoxia mediates cancer hallmarks and results from reduced oxygen level due to irregularities in tumor vascularization or when the tumor size prevents oxygen diffusion and triggers angiogenesis to compensate for low oxygen. Cancer stem cells (CSCs) are a rare subpopulation, able to self-renew and to give rise to tumor-initiating cells. It is proposed that CSCs' secretions help to recruit endothelial cells via angiogenic factors to establish tumor vascularization. In the tumor microenvironment, the effect of hypoxia on CSCs and the impact of their secretions on triggering angiogenesis and tumor vascularization remain questionable. In this study, three-dimensional (3D) CSCs derived from MCF-7 were directly exposed to repetitive long-term cycles of hypoxia to assess its effect on CSCs and then to evaluate the role of the hypoxic CSCs' (CSCsHYP) secretions in angiogenesis using (HUVECs) as a model for tumor neovascularization response. METHODS: CSCs derived from MCF-7 cell-line were expanded under repetitive, strictly optimized, long-term/continuous and intermittent hypoxic shots for almost four months to assess hypoxic effect on CSCs, sorted based on CD44+/CD24- biomarkers. Hypoxic phenotype of CSCsHYP was evaluated by assessing the acquired chemoresistance using MTT assay and elevated stemness properties were assessed by flow cytometry. To evaluate the effect of the secretions from CSCsHYP on angiogenesis, HUVECs were exposed to CSCsHYP conditioned-medium (CdM)-in which CSCs had been previously grown-to mimic the tumor microenvironment and to assess the effect of the secretions from CSCsHYP on the HUVECs' capability of tube formation, migration and wound healing. Additionally, co-culture of CSCsHYP with HUVECs was performed. RESULTS: CSCsHYP acquired higher chemoresistance, increased stemness properties and obtained greater propagation, migration, and wound healing capacities, when compared to CSCs in normoxic condition (CSCsNOR). HUVECs' tube formation and migration abilities were mediated by hypoxic (CSCs) conditioned media (CdM). DISCUSSION: This study demonstrates that chemoresistant and migrational properties of CSCs are enhanced under hypoxia to a certain extent. The microenvironment of CSCsHYP contributes to tumor angiogenesis and migration. Hypoxia is a key player in tumor angiogenesis mediated by CSCs.

Phytochemical analyses and antibacterial activities of Erodium, Euphorbia, Logoecia and Tamarix species.

INTRODUCTION: Resistance against commonly used antibacterial agents has become a globally recognized threat to human health. Therefore, the development of new and effective antibacterial agents is necessary to treat infections caused by resistant bacterial strains; plants are a promising source of new agents to be tested. METHODOLOGY: The minimum inhibitory concentrations (MIC) of ethanolic extracts of Erodium gruinum, Euphorbia hierosolymitana, Logoecia cuminoides, and Tamarix tetragyna against 10 Gram-negative and 5 Gram-positive bacteria were determined using agar well diffusion and microtiter plate dilution methods, respectively. The phytochemical composition of the crude extracts of the plants was determined using HPLC. RESULTS: Pseudomonas aeruginosa ATCC 27853, Klebsiella pneumoniae, Proteus mirabilis, and Acinetobacter baumannii were sensitive to E. gruinum and E. hierosolymitana extracts. P. aeruginosa ATCC 27853 and M. catarrhalis were sensitive to L. cuminoides extract. P. aeruginosa ATCC 27853, P. mirabilis, and K. pneumoniae were sensitive to T. tetragyna extracts. For Gram-positive bacteria, Staphylococcus aureus ATCC 33591 and ATCC 43300 were sensitive to E. gruinum and E. hierosolymitana extracts. S. aureus ATCC 43300 and ATCC 33591 and Group D Streptococcus were sensitive to T. tetragyna extract. All Gram-positive bacteria were completely resistant to the extract of L. cuminoides. The major phytochemical components of the plant extracts belonged to flavonoids, tannins, terpenes, quinones, phytosterols, phytoestrogens, carbohydrates, fatty acids, and coumarin. CONCLUSION: The study showed the potential of the development of antibacterial agents from these plants. Phytochemical analysis revealed compounds that are candidates for new antibacterial drugs.