This research was aimed to the purification and characterization of cytosine deaminase as a medically important enzyme from locally isolated Escherichia coli; then studying its cytotoxic anticancer effects against colon cancer cell line. Cytosine deaminase was subjected to three purification steps including precipitation with 90% ammonium sulfate saturation, ion exchange chromatography on DEAE-cellulose column, and gel filtration chromatography throughout Sephadex G-200 column. Specific activity of the purified enzyme was increased up to 9 U/mg with 12.85 folds of purification and 30.85% enzyme recovery. Characterization study of purified enzyme revealed that the molecular weight of cytosine deaminase produced by E. coli was about 48 KDa, the highest enzyme activity at pH 8.5, and is most stable at pH 7.5 - 9, the enzyme also showed a full activity at a range of temperatures between 45-60 0C. Enzyme activity was strongly inhibited in the presence of mercuric chloride and copper sulphate, when added individually at a constant concentration. However, calcium chloride, manganese chloride and ferric chloride caused a little increase in enzyme activity while sodium azide had no effect on enzyme activity. Upon cytotoxic effect study through micro-cultured tetrazolium assay (MTT) against Caco-2 cell line. Purified cytosine deaminase was found to inhibit the growth of Caco-2 cancer cell line with an IC50 of 242.5 ?g/ml in a comparison to an IC50 of 1864 ?g/ml for crude enzyme. Besides, the enzyme didn’t show significant effect on WRL normal cell line.
The aim of this research to show the role of some enzymes in pathological mechanism of rheumatoid arthritis (RA) disease. Sixty patients with RA and matched number of apparently healthy volunteers were included in the study. Spectrophotometric methods were used to determine Peroxy nitrite (ONOO), Nitric oxide (NO), Nitric oxide synthase activity (NOS) cycloxygenase-2 activity (COX-2), glutathione peroxidase (GPX) activity and superoxide dismutase (SOD) activity in serum of both groups. Colorimetric assay kits were used to determine Iron. Rheumatoid factor (RF) was determined using Imuno-Latex kit. ONOO, NO levels, and NOS activity were significantly higher in the patients compared to the control group. Conversely, Iron level, SOD
... Show MoreStimulative effect of 10 mW He-Ne laser on the phagocytic activity of human polymorphonuclear leukocytes( PMNs) has been studied in vitro. Normal polymorphonuclear leucocytes were isolated from the human peripheral blood. A mixture of 0.25 ml of Hanks solution, 0.25 ml of serum, 0.25 ml of Candida albicans suspension and 0.25 ml of PMNs suspension was prepared. The samples of mixture of PMNs and Candida were subdivided in 1 ml ependrof tubes and irradiated to He-Ne laser for 1, 3, 5, 10 and 20 min. The diameter of the irradiated area was 0.8 cm. For calculation of Phagocytic index before and after irradiation, the samples were incubated (37°C) at 5, 15, 30, 60 min. The slides of samples were prepared and stained using Giemsa stain. The
... Show MoreBackground: The potential use of zinc oxide and other metal oxide nanoparticles in biomedical are gaining interest in the scientific and medical communities, largely due to the physical and chemical properties of these nanomaterials. The present work revealed the effect of zinc oxide nanoparticles (ZnONPs) on the total salivary peroxidase enzyme activity of human saliva in comparison to de-ionized water. Materials and methods: Forty eight unstimulated saliva samples were collected from dental students/University of Baghdad 18-22 years. Then measure the total salivary peroxidase activity first without any addition to human saliva as a control, second with dilution the saliva with de-ionized water, and third with zinc oxide nanoparticles in c
... Show MoreThe present study was performed to evaluate the anti-fungal effect of alcoholic extract of Solanum nigrum (AESn) on the growth of Microsporum canis, the causes agent of ring worm. The results of this work referred to the inhibitory effect of the studied extract on the growth of tested fungi. The percentages of inhibition were (7.88 %, 19.88%, 23.41%, 57.65%), in comparison to the control, when (2%, 4%, 6%,8% ) of tested extract were used, respectively. The data illustrated that the higher concentrations of the extract are applied, the more inhibition of fungal growth is produced.
In this work, Schiff base ligands L1: N, N-bis (2-hydroxy-1-naphthaldehyde) hydrazine, L2: N, N-bis (salicylidene) hydrazine, and L3:N –salicylidene- hydrazine were synthesized by condensation reaction. The prepared ligands were reacted with specific divalent metal ions such as (Mn2+, Fe2+, Ni2+) to prepare their complexes. The ligands and complexes were characterized by C.H.N, FT-IR, UV-Vis, solubility, melting point and magnetic susceptibility measurements. The results show that the ligands of complexes (Mn2+, Fe2+) have octahedral geometry while the ligands of complexes (Ni2+) have tetrahedral geometry.
Four electrodes were synthesized based on molecularly imprinted polymers (MIPs). Two MIPs were prepared by using the diclofenac sodium (DFS) as the template, 2-hydroxy ethyl metha acrylate(2-HEMA) and 2-vinyl pyridine(2-VP) as monomers as well as divinyl benzene and benzoyl peroxide as cross linker and initiator respectively. The same composition used for prepared non-imprinted polymers (NIPs) but without the template (diclofenac sodium). To prepared the membranes electrodes used different plasticizers in PVC matrix such as: tris(2-ethyl hexyl) phosphate (TEHP), tri butyl phosphate (TBP), bis(2-ethyl hexyl) adipate (BEHA) and tritolyl phosphate (TTP). The characteristics studied the slop, detection limit, life time and linearity range of DF
... Show MoreIn this work lactone (1) was prepared from the reaction of p-nitro phenyl hydrazine with ethylacetoacetate, which upon treatment with benzoyl chloride afforded the lactame (2). The reaction of (2) with 2-amino phenol produced a new Schiff base (L) in good yield. Complexes of V(IV), Zr(IV), Rh(III), Pd(II), Cd(II) and Hg(II) with the new Schiff base (L) have been prepared. The compounds (1, 2) were characterized by FT-IR and UV spectroscopy, as well as characterizing ligand (L) by the same techniques with elemental analysis (C.H.N) and (1H-NMR). The prepared complexes were identified and their structural geometries were suggested by using elemental analysis (C.H.N), flame atomic absorption technique, FT-IR and UV-Vis spectroscopy, in additio
... Show MoreComplexes of Au(III) ,Pd (II) , Pt (IV ) and Rh(III) with S – propynyle -2- thiobenzimidazole (BENZA) have been prepared and characterized by IR and UV- Visible spectral methods in addition to magnetic and conductivity measurements and micro – elemental analysis (CHN).The probable structures of the new complexes have been suggested.