QUALITATIVE PHYTOCHEMICAL COMPARISON BETWEEN FLAVONOIDS AND PHENOLIC ACIDS CONTENTS OF LEAVES AND FRUITS OF MELIA AZEDARACH (FAMILY: MELIACEAE) CULTIVATED IN IRAQ BY HPLC AND HPTLC

Objective: The aim of our study was to compare between flavonoids and phenolic acids contents of leaves and fruits of Melia azedarach since no phytochemical investigation had done previously in Iraq.Methods: The leaves and fruits of Melia azedarach were extracted by soxhlet using 80% ethanol then the dried extract was suspended in water and fractionated using petroleum ether, chloroform, ethyl acetate, and n-butanol. The n-butanol fraction was hydrolyzed by acid and partitioned with ethyl acetate. The different fractions containing flavonoids and phenolic acids were analyzed by HPLC and HPTLC.Results: The HPLC results revealed the presence catechin-7-O-glycoside in fruit only, while kaempferol-7-O-glycoside is found in the leaves only. Catechin and its glycosides are more abundant in the fruits than in the leaves. The HPTLC results revealed that kaempferol and quercetin are present in all fractions of leaves and fruits as aglycones and as glycosides. Free chlorogenic was found in both leaves and fruits.Conclusion: No major differences were found between the flavonoids and phenolic acids contents of the leaves and fruits of Melia azedarach

Differential expression patterns of leukaemia associated genes in leukaemia cell lines compared to healthy controls

Introduction: The phenotype and genotype of cancer cells portray hallmarks of cancer which may have clinical value. Cancer cell lines are ideal models to study and confirm these characteristics. We previously established two subtracted cDNA libraries with differentially expressed genes from an acute myeloid leukaemia patient with poor prognosis (PP) and good prognosis (GP). Objective: To compare gene expression of the leukaemia associated genes with selected biological characteristics in leukaemia cell lines and normal controls. Methodology: Expression of 28 PP genes associated with early fetal/embryonic development, HOX-related genes, hematopoiesis and aerobic glycolysis/hypoxia genes and 36 GP genes involved in oxidative phosphorylation, protein synthesis, chromatin remodelling and cell motility were examined in B-lymphoid (BV173, Reh and RS4;11) and myeloid (HL-60, K562) leukaemia cell lines after 72h in culture as well as peripheral blood mononuclear cells from healthy controls (N=5) using semi-quantitative polymerase chain reaction (PCR) method. Cell cycle profiles were analysed on flow cytometry while MTT cytotoxicity assay was used to determine drug resistance to epirubicin. Results: Genes expressed significantly higher in B-lymphoid leukaemia cell lines compared to healthy controls were mostly of the GP library i.e. oxidative phosphorylation (3/10), protein synthesis (4/11), chromatin remodelling (3/3) and actin cytoskeleton genes (1/5). Only two genes with significant difference were from the PP library. Cancer associated genes, HSPA9 and PSPH (GP library) and BCAP31 (PP library) were significantly higher in the B-lymphoid leukemia cell lines. No significant difference was observed between myeloid cell lines and healthy controls. This may also be due heterogeneity of cell lines studied. PBMC from healthy controls were not in cell cycle. G2/M profiles and growth curves showed B-lymphoid cells just reaching plateau after 72 hour culture while myeloid cells were declining. IC50 values from cytotoxicity assay revealed myeloid cell lines had an average 13-fold higher drug resistance to epirubicin compared to B-lymphoid cell lines. Only CCL1, was expressed at least two-fold higher in myeloid compared to B-lymphoid cell lines. In contrast, MTRNR2, EEF1A1, PTMA, HLA-DR, C6orf115, PBX3, ENPP4, SELL, and IL3Ra were expressed more than 2-fold higher in B-lymphoid compared to myeloid cell lines studied here. Conclusion: Thus, B-lymphoid leukaemia cell lines here exhibited active, proliferating characteristics closer to GP genes. Higher expression of several genes in B-lymphoid compared to myeloid leukaemia cell lines may be useful markers to study biological differences including drug resistance between lineages

Analysis of peripheral blood of ovarian cancer patients indicates higher sub-populations of natural killer and B cells compared to healthy volunteers

Ovarian cancer is a challenging disease to treat, and one of the potential treatments is by immunotherapy. NK cells have been shown to play a role in slowing tumour progression and cancer development. This study aims to investigate the numbers of NK cells and other lymphocyte sub-populations in ovarian cancer and their impact on ovarian cancer clinical outcome. This project aims to study the significance of different lymphocyte populations, particularly NK cells, involved in the peripheral blood of ovarian cancer patients. Venal blood was drawn from ovarian cancer patients before chemotherapy. PBMCs were isolated from 13 ovarian cancer patients and 11 age-matched healthy volunteers. Immunophenotyping was performed using a commercial kit to quantify the lymphocyte populations and RNA isolation performed to examine the expression of KIR genes using reverse transcription polymerase chain reaction. Immunophenotyping of PBMC was successfully performed on 13 ovarian cancer patients and 11 healthy controls. Significant increases in the mean of peripheral NK cells and B cells were found in ovarian cancer patients as compared to the healthy controls (P=0.0559). No other significant results were obtained for C D4 and CD8 lymphocytes. There was significant increase in numbers of NK cells and B cells in ovarian cancer patients as compared to the healthy volunteers. These results should be pursued with a larger sample size with the hopes of finding a significant difference between the two groups and to provide a keener insight into are promising preliminary results the immune defence against ovarian cancer

Evaluation of the Genotoxicity of the Aerial Parts of Iraqi Euphorbia cyathophora on Bone Marrow and Spleen Cells in Mice

The aim of the study was extraction of arial part of Euphorbia cyathophora constituents with methanol and evaluate its effect on mitotic index and total chromosomal aberration bone marrow cell and spleen cell in mice  200 gm of E. cyathophora fine powder was defatted then extracted by cold maceration 80% ethanol for seven days. The extract was filtered and dried in a rotary evaporator then the dried extract was suspended with water and consecutively extracted using chloroform, ethyl acetate for each. The aqueous layer was then mixed with 100ml methanol. These fractions are dried under reduced pressure to obtain the dry extract. Twenty-four Albino mice were used for the experiment. The animals were divided into four groups: Group 1: Mice were treated with distilled water. The dose was given daily for seven successive days. Group 2: Mice were treated with a single dose (20mg/kg) of methotrexate (positive control). Group 3: Mice were treated with (100mg/kg) of menthol fraction for seven successive days. Group 4: Mice were treated with (200mg/kg) of methanol fraction for seven successive days. Mice were sacrificed by (spinal dislocation). Samples of bone marrow cells and spleen cells were taken and genotoxic analyses. Methanol fraction of Euphorbia cyathophora at a dose of 100mg/kg demonstrated a significant decrease in mitotic index and a significant increase in total chromosomal aberrations as compared to distilled water in both bone marrow cells and spleen cells (p<0.05). 100 mg/kg and 200 mg/kg of methanol fraction of Euphorbia cyathophora that showed to be significantly higher in mitotic index and significantly lower in total chromosomal aberration as compared to methotrexate (p<0.05). In conclusion the current study revealed that the methanol fraction of aerial parts of Euphorbia cyathophora is genotoxic but its genotoxicity is less than that of methotrexate

ADAMTSL5 and CDH11: putative epigenetic markers for therapeutic resistance in acute lymphoblastic leukemia

Background and objectives: DNA hypermethylation has been linked to poor treatment outcome in childhood acute lymphoblastic leukemia (ALL). Genes differentially methylated in the chemoresponsive pre-B-ALL compared to chemoresistant pre-B-ALL cases provide potential prognostic markers. Methods: DNA methylation profiles of five B-ALL childhood patients who achieved morphological complete remission (chemoresponsive) and five B-ALL patients who did not (chemoresistant) after induction treatments as well as four normal controls were compared on 27 000 CpG sites microarray chips. Subsequently, methylation-specific polymerase chain reaction (MSP) on selected hypermethylated genes was conducted on an additional 37 chemoresponsive and 9 chemoresistant B-ALL samples and 2 normal controls. Results: Both methods were found to be highly correlated. Unsupervised principal component analysis showed that the chemotherapy-responsive and -resistant B-ALL patients could be segregated from one another. Selection of segregated genes at high stringency identified two potential genes (CDH11 and ADAMTSL5). MSP analysis on the larger cohort of samples (42 chemoresponsive, 14 chemoresistant B-ALL samples and 6 normal controls) revealed significantly higher rates of hypermethylation in chemoresistant samples for ADAMTSL5 (93 vs. 38%; p = 0.0001) and CDH11 (79% vs. 40%, p < 0.01). All control cases remained unmethylated. Conclusion: Chemoresistant B-ALL patients are associated with increased methylation in ADAMTSL5 and CDH11. These findings need to be validated in a larger group of patients, and the functional biological and prognostic significance of differential methylation needs to be studied further

Assessment of P-gp and MRP1 activities using MultiDrugQuant Assay Kit : a preliminary study of correlation between protein expressions and its functional activities in newly diagnosed acute leukaemia patients.

Multidrug resistance (MDR) is believed to be responsible for poor response of patients towards chemotherapy particularly patients with acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL). The best-characterized resistance mechanism is the one mediated by permeability-glycoprotein (P-gp) encoded by MDR1 gene, which is responsible for drug efflux. We studied P-gp and multidrug resistance-associated protein 1 (MRP1) expression and functional activities in 43 newly diagnosed acute leukemia cases (19 paediatric ALL cases and 24 adult AML cases). The expression and functional activities were examined using flow cytometry and MultiDrugQuant assay kit (involving calcein AM uptake and efflux). P-gp and MRP1 expression and its functional activities were observed in 68.4% of paediatric ALL. In adult AML cases, all cases expressed MRP1 and its functional activities but only 58.3% were positive for P-gp and its functional activities. We were able to show a significant correlation between the expression of the multidrug resistant protein (P-gp and MRP1) and their functional activity in adult AML and paediatric ALL samples

Comparison of decompression alone versus decompression with fusion for stenotic lumbar spine: A systematic review and meta-analysis

The first line of treatment for lumbar spinal stenosis (with or without lumbar degenerative spondylolisthesis) involves conservative options such as anti-inflammatory drugs and analgesics. Approximately, 10%-15% of patients require surgery. Surgical treatment aims to decompress the spinal canal and dural sac from degenerative bony and ligamentous overgrowth. Different studies have given conflicting results. The aim of our study is to clear the confusion by comparing two surgical techniques. This meta-analysis was conducted in accordance with the preferred reporting items for systematic reviews and meta-analysis (PRISMA) guidelines. A literature search was conducted of the Ovid Embase, Scopus, Pubmed, Ovid Medline, Google Scholar, and Cochrane library databases. A quality and risk of bias assessment was also done. The analysis was done using Revman software (The Nordic Cochrane Centre, The Cochrane Collaboration, 2014, Copenhagen, Denmark). A total of 76 studies were extracted from the literature search and 29 studies with relevant information were shortlisted. Nine studies were included in the meta-analysis after a quality assessment and eligibility. Fusion with decompression surgery was found to be a better technique when compared to decompression alone for spinal stenosis in terms of the Oswestry Disability index and the visual analog pain scale for back and leg pain. On the basis of the meta-analysis of the recent medical literature, the authors concluded that decompression with fusion is a 3.5-times better surgical technique than decompression alone for spinal stenosis

Phytochemical study and thin layer chromatography of Ficusreligiosa leaves extract cultivated in Iraq

Ficusreligiosa Linn, (Moraceae), is a large evergreen or deciduous, irregularly shaped tree. Traditionally the leaves are used for the treatment of constipation, vomiting, hiccup, and others. Leaves were extracted by two methods; maceration and soxhelt using hexane and   80% aqueous methanol, then subjected to preliminary phytochemical examination, fractionation with chloroform, ethyl acetate, and n.butanol, then TLC. Soxhelt was the suitable extraction method. Sterols, alkaloids, saponins, tannins, and flavonoids were identified in leaves. TLC examination demonstrates the possible presence of stigmasterol, chlorogenic acid, caffeic acid, rutin, and luteolin or apigenin

Overview of Metal Ions Released from Fixed Orthodontic Appliance: In Vitro Studies

Intraoral fixed orthodontic appliances are frequently used in the clinical practice of dentistry. They are made from alloys containing different metals at various percentages. The use of these appliances leads to the long-term exposure of patients to these materials, and the potential toxic effects of this exposure raises concerns about patient safety. Thus, the biocompatibility (corrosion behaviour and toxicity) of these materials has to be evaluated prior to clinical use. In the present report, the most recent studies in the scientific literature examining metal ion release from orthodontic appliances and the toxic effects of these ions have been reviewed with a special focus on cytotoxicity and genotoxicity. Previous studies suggest that a case-by-case safety evaluation is required to take into account the increasing variability of materials, their composition and the manufacturing processes. Moreover, in vitro toxicity studies in regard to metal release, cytotoxicity and genotoxicity are still scarce. Therefore, in vitro monitoring studies are needed to establish cause-effect relationships between metal ion release and biomarkers of cytotoxicity and genotoxicity. Further investigations could be performed to elucidate the toxic mechanisms involved in the observed effects with a special emphasis on oxidative damage