Journal of Biotechnology Research Center

Abstract:
The study included manufacturing of fermented dairy products by using full cream milk of four kinds of mammalian: (Buffalos, Cows, Sheep's and Goats); with the use of Lactobacillus casei as a starter for the production of fermented dairy products which had the ability to lower cholesterol percentage in the above mentioned products by (71.4, 70, 74.8 and 67.7)% respectively. The viability of Lb. casei had not been affected significantly during storage shelf life of 21days " The product shelf life " , keeping their therapeutic properties unaltered with high viable number of bacteria at time of consumption. The viable counts of the bacteria after storage period for manufactured products were (1.06× 109, 8.1× 108, 7.5× 108and 8× 108) CFU/ml respectively. These numbers represent a decrease equal to one logarithmic cycle for each of manufactured products of Cows, Sheep's and Goats milk, and the decrease of bacteria's viability of manufactured products of Buffalos milk was less than one logarithmic cycle. Results of statistical analysis showed that there was highly significant differences (P<0.05) in the viable bacterial cells counts between manufactured products. By sensory comparison of the manufactured fermented products together, the results shows that the manufactured products from Buffalos milk was the best then the manufactured products of Cows milk then Sheep's milk then goats milk

Abstract
Samples of bovine pancreatic and calf thymus glands were collected from local slaughterers to extract deoxyribonuclease I enzyme (DNase I) and DNA respectively. Crude extract was prepared by using cooled distilled water and 0.25 N sulfuric acid .This study show that the best condition for the crude extract activity was 17 U/ml and 1 hr. Incubated period reaction with its substrate.The bovine pancreatic DNase I was purified by several steps of precipitation using ammonium sulphate to 529.4 times, with an enzymes yield 6.35%. Enzyme characterization studies indicate that: it is active at 2.7 U/ml, and 50 µg /ml DNA after 30 min of reaction time, the enzyme activity was higher at pH 6.5 and it showed stability at pH 9. The maximum enzyme activity was reported at 50 °C The results obtained from the role of metal ions (Mg+2, Mn+2) on enzyme activity indicate that these ions stimulated the enzyme activity while the Ca+2 and Cu+2 had lower stimulating activity on the enzyme but Ag+1 and Hg+2 showed inhibitory effect on enzyme activity. In addition the enzyme activity was inhibited by using denaturing Sodium dodecyl sulphate (SDS), chelating agents Ethylene diamine tetra acetic acid (EDTA), and reducing agents (2-Merceptoethanol and Urea) , and maintained it's activity when incubated with Phenyl Methyl Sulphonyl Floride (PMSF) .

Abstract
The effect of root exudates of fifty Sorghum genotypes on seed germination and seedling growth of millet were studied in vitro. The fifty genotypes were divided into four groups according to their effect on percentage reduction in root and shoot and whole plant average lengths of millet seedling growth. The first group caused slight stimulation, the second slight inhibition, the third gave limited inhibition and the fourth caused high inhibition. In seedling growth. Five genotypes of sorghum were selected, one from group two and four from group four. These five genotypes were cultured in the field in Autumn 2007 and at the end of the season plants were removed, dried and grind. Three concentrations of each of these genotypes powder were prepared, in addition to the control treatment, (0.0, 3.0, 6.0 and 9.0) g/kg soil to study their effects on the growth of Amaranthus retroflexus which cultured in pots and kept in experimental field. Data on number of leaves, lengths of shoot and root and whole plant were taken as well as wet and dry weight were measured. Percentage of Amaranthus retroflexus seeds germination were evaluated in an experiment cultured in Petri dishes containing concentrations (0.0, 5.0 and 10.0%) of aqueous extracts of each of the five genotypes powder. Results showed high significant inhibition for the four genotypes from group four in comparison with the one genotype from group two. Significant differences among these genotypes in their inhibition on such parameters as shoot, root, whole plant growth, number of leaves, and wet and dry weight. The genotype 2005-K-Type 1050 gave the highest reduction in plant length 53.2% incomparing with the genotype 2005-K-Type 1033 which gave 0.7%. The percentage of seed germination of Amaranthus retroflexus was significantly reduced as the concentration of the aqueous extract increased and there were significant differences among genotypes on this parameter. The concentration of the powder of each genotype added to the soil produced significant reduction in all parameters studied and the effect increased as the concentration increased in soil. This research discusses the potential of the allelopathic effect of some Sorghum genotypes on weed germination and growth reduction and the possibility of employing them in weed control program with the intention to use less herbicide.

Abstract
This study was carried out to separate acute myeloid leukemia (AML) blast cells and studies their proliferation in short-term culture. The separation procedure include three steps; Ficoll gradient separation, depletion of macrophages and depletion of (lymphocytes and of monocytes) for preparation of highly pure native AML blast cells from blood samples collected from patients with moderate blast percentage. Results showed that this procedure is an inefficient due to a decrease in total cell number and contamination with other cells after each separation step. Proliferation of native AML blast cells in short term-culture by cultivating isolated AML blast cells in RPMI medium supplemented with 20% human plasma at a concentration 1×106/ml in the presence and absence of colony -stimulating factor which was provided by conditioned media (PHA-leucocytes-, plasmacytoma cell line- and Hep-2 conditioned medium. The effect of each conditioned medium on proliferation of AML blast cells was studied separately. Results showed that plasmacytoma cell line conditioned medium didnot stimulate the proliferation of native AML blast cells, while cells seeded on media containing 10%PHA-LCM showed an increase in cell number and growth of the cells was observed for approximately 3 days and then decreased.

Abstract
Genomic instability resulting in multiple molecular events believed to be a driving force in the carcinogenic process. In this study, the random amplified polymorphic DNA (RAPD) technique, a simple polymerase chain reaction (PCR) based DNA polymorphism assay system was used for analyzes genomic instability in blood and tissues obtained from five breast cancer patients. DNAs from breast cancer tissues and corresponding DNAs from blood samples were amplified by RAPD with five different 10-bases arbitrary primers. The ability to detect genomic instability in five cancer tissues by each single primer ranged from 60 to 100 percent. Samples 3, and 4 represented the highest genomic instability since it detected by all primers used. Changes in the genome that were revealed by RAPD included deletion or insertion, and allelic losses or gains. The most important finding that emerged from the RAPD analysis, is that deletion was also observed in blood DNAs which revealed by the absence of amplified DNA fragments from blood DNAs, while the corresponding fragments were present in the tumor DNA. Our results display that an insertion of a 468bp amplified fragment was observed in 3 of 5 tumor samples (two fragments using primer OPA-03 and the third fragment using primer OPA-10) whereas, the same 468bp amplified fragment was deleted in 2 of 5 blood samples one using the primer OPA-03 and the second fragment using primer OPA-13. Genomic instability analyzed by RAPD is important to understand the molecular events in breast cancer.

Abstract
Trichothecenes are natural secondary metabolites causing economic losses and health hazard to human and farm animals which are produced by several species of Fusarium and some other genera on different agricultural commodities. Study on trichothecenes mycotoxicosis revealed morphological, biochemical, and histopathological changes. After intraperitoneal injection of the toxin in male mice with different concentration for 35 days shows marked increase in body weight, dyspanea, shivering, bristling up of hair, hair falling, anomalies of eyes, and irritation around neck, also abdominal hemorrhage and clot accumulation in abdomen. In addition to inclusion (retention) cyst forms on liver. The biochemical studies on liver function by measuring GPT and GOT enzymes level have been done. An increase level of these enzymes in treated animal in comparison with control animal which indicating abnormal function of liver observed. The histopathological study on sections from the liver of treated animal with trichothecenes revealed many alterations in liver which includes congestion, kupffer cells hyperplasia, dilated sinusoids and mononuclear cells infiltration around the portal area.

Abstract
Approximately, 50% of the dry mass of the outer membrane of gram-negative bacteria consists of proteins, and more than 20 immunochemically distinct proteins (termed outer membrane proteins [OMPs]) have been identified. An identified local strain of Klebsiella pneumoniae was used as a primary source for the isolation and purification of porins. Multiple concentrations of purified porins (5, 10, 15, 20, 25) g/ml were incubated with three different cell lines for (24, 72 , 120) hrs, after the end of the incubation periods, the cells were treated with Cell proliferation ELISA, BrdU (colorimetric) kit to evaluate the antiproliferative effects of porins. The results revealed that porins are potent antiproliferative agent in a time and concentration dependent manner and thus could greatly affect prokaryote-eukaryote interaction as well as the whole inflammatory process resulted after infection with gram negative bacteria

Abstract
This study aimed to evaluate the effect of aqueous extract of sage (Salvia officinalis) on bone marrow cells in albino male mice by using three doses ( 83.9, 167.8 or 251.7 mg/kg) and cytosar drug at dose of ( 1.54 mg/kg). The results showed that sage has the ability to increase the mitotic index in mice in comparing with the negative control and in mice treated with cytosar drug that caused reduction in mitotic index. The results of pre- and post-treatment with the ideal dose of aqueous extract of sage and cytosar drug showed the ability of sage to increase the mitotic index of bone marrow cells in mice in comparing with the negative controls.

Abstract
Urinary tracts infection remains a common troublesome health problem in the world. Although E.coli is normal intestinal flora but considered as the main important opportunistic active uropathogene because of it’s pathogenesity which referred to it’s different virulence factors like hemolysin, biofilm, enterotoxins, shiga like toxins, cytotoxic necrotizing factor and others. Seventy seven E.coli isolates which isolated from 125 midstream urine samples of patients suffering from different types of UTIs. The results showed that 44 isolates 57% were produced hemolysin. These isolates were differed in the efficiency of erythrocyte lysis because which depend on type of hemolysin and source of blood. Seventy two isolates 90.9% were formed biofilm with variety in thickness of biofilm layer. Each isolate produce hemolysin also formed biofilm. Presence of genes encoded for virulence traits of UPEC (hemolysin, biofilm) were examined by PCR with specific primers. The results showed that the percentage of α-hly gene encoded for α-hemolysin 50% while percentage of eae gene encoded for intimin and E-hly gene encoded for enterohemolysin were 40% and 20% respectively.

Abstract:
In this study the extracts of the Iraqi herb Tribulus terrestris (Al-Hassage or Al-Kutub) was done by using of polar and non polar solvents, then the biological activity of these extractants was studied in two field. First, the antibacterial activity invitro on gram positive bacteria (Staphylococcus aureus), and gram negative bacteria (E. coli, Proteus vulgaris, Pseudomonas aerugiuosa, and Klebsiella pneumonia), all extracts showed considerable activity against all bacteria. Second, the effect of extracts on free serum testosterone level in male mice invivo, the alcoholic, and acetonitrilic extracts showed significant (P < 0.05) increase in free serum testosterone level, and we found that the extracts contained compounds with less genotoxic effects in mice germ cells.

Abstract
Nicotinamide adenine dinucleotide phosphate-dependent isocitrate dehydrogenase (NADP+-IDH; EC 1.1.1.42) was extracted from red kidney beans (Phaseolus vulgaris.) after the beans were placed into Murashige-Skoog medium and incubated under continuous white light (110 μmol photon m−2 s−1), then filtered, centrifuged and the supernatant was used for purification. The enzyme purified using ammonium sulphate precipitation, DEAE-cellulose and Matrex Bio red A (dye-ligand-chromatography) techniques, and exhibits several bands through electrophoresis, with one band corresponds to the IDH activity. Km values for the enzyme was 55.71± 4.56 x 10-6M. The enzyme has an optimum pH at 8.5, and optimum temperature at 30°C. The enzyme can be stable at RT (about 25°C) for 180min, but the activity disappears at 400min. Enzyme activity appears to be independent of divalent metals in deionized water, but the addition of Mg+2 and Mn+2 by 4.5 and 2-folds respectively. The purified enzyme was injected into white rabbits to raise an antiserum against NADP+-IDH. The specificity of the antiserum was assayed by its ability to decrease the NADP+-IDH activity present in the extract. NADP+-IDH activity decreased when the extract was incubated with increasing volumes of the antiserum obtained.

Abstract:
This work involves the chemical synthesis of novel complexes derived from steroid hormones using testosterone and methandrostenolon as starting materials. When these starting materials react with thiosemicarbazide, L1 (testosteronthiosemicarbazone ) and L2 (methandrostenolonthiosemicarbazone ) are formed, and when they react with Cr (III), Co (II), Ni (II), and Cu (II) metal ions a new complexes are formed. The chemical structures for all the prepared compounds were characterized by elemental analysis, FT-IR, and UV/visible spectra. Moreover molar ratio M:L, metal content M%, and magnetic moments (μeff.) were also determined. The IR spectral data suggest the involvement of sulphur and azomethane nitrogen in coordination to the central metal ion. The free ligands and their metal complexes have been tested in vitro against a number of microorganisms (gram positive bacteria (Staphylococcus aureus), and gram negative bacteria (E. coli, Proteus vulgaris, Pseudomonas, and Klebsiella) in order to assess their antimicrobial properties. All the prepared complexes showed considerable activity against all bacteria.

Abstract:
The aim of this work is to determine the duration of goat cell cycling in vitro.Goat peripheral blood lymphocytes were grown in RPMI-1640 medium containing bromodeoxyuridine (BrdU 10 μg/ml) for 72 h. Blastogenic index (BI), mitotic index (MI), cell cycle progression (CCP) and sister chromatid exchanges (SCE) were determined. Cultured lymphocytes from, whole blood or from leukocyte rich plasma in RPMI-1640 medium containing BrdU showed little differences in BI, MI, but significant differences were seen in cell cycle progression. BI, MI, and CCP from different goat breed were compared. Also, the percentage of lymphocyte blastogenesis, mitoses and cell cycle progression from goat, were compared to those from sheep, and human whichgrown under similar conditions. On successive incubation periods, the cell cycle duration of blood lymphocytes was determined through the mitotic activity. The cells reached first, second and third mitoses after 25, 40 and 48 h, post incubation respectively. Sub culturing of growing lymphocytes was performed from 3 to 45 days to obtain a lymphoblastoid cells. The characterization of their differentiation is required Establishment of goat blood lymphocyte culture will help in gene marker’s detection in their somatic cells.

Abstract
The present study was designed to investigate the role of P. odoratissimum aquatic extracts in reducing the genotoxic effects of metronidazole in mice in vivo and human blood lymphocyte in vitro. The parametrers which evaluated in mice were used: mitotic index and chromosomal aberrations in bone marrow, while for human blood lymphocyte were mitotic index, blast index, replicative index, sister chromatid exchange and chromosomal aberrations. The cytogenetic effects of the drug and plant aquatic extracts were investigated after four days of oral administration for mice with metronidazole and aqueous extract at doses 400mg/kg and 100 mg/kg respectively while the concentrations of metronidazole and aqueous extract in human blood lymphocyte culture was 80µg/ml, and 10µg/ml respectively. An interaction study of plant extract with metronidazole was carried out through three types of treatments (before, after and mixture of plant extract and drug treatment) to determine the activity of P. odoratissimum aqueous extract in reducing the side effects of drug both in vitro and in vivo. Aquatic extract of P.odoratissimum at the concentration of 10µg/ml, showed a protective value against the genotoxic effect of metronidazole at 80µg/ml. concentration .In mouse bone marrow cells and human blood lymphocyte culture, this was more pronounced in pre-treatment and simultaneous treatment than in post-treatment. So P. odoratissimum aquatic extract is considered as desmutagen in the first order and bioantimutagen in the second order, as a result for its ability to repair CA and increase MI in mouse system and in human blood lymphocyte culture system . It also had the ability to increase BI and RI and decrease SCE in human blood lymphocytes culture in vitro.

Abstract
The present study was carried out with the aim to evaluate the hematological and cytogenetic effects of seed aqueous extract of the plant Ammi majus (0.5, 1.0, 1.5) mg/kg in albino male mice. The investigated parameters were total count of leucocytes (TLC), mitotic index (MI), micronucleus (MN) formation and chromosomal aberrations. The mitomycin C (MMC) was used as a mutagen in the interaction with the plant extract (pre- and post-treatment), with the aim to determine the antimutagenic efficiency of the plant extract, and in all cases, the materials were given orally. In the first treatment, the results indicated that the dose 1.5 mg/kg of the extract enhanced the parameters investigated and a significant increase was observed in TLC (10070 cells/cu.mm.blood) as compared to negative (7290 cells/cu.mm.blood) or positive (4910 cells/cu.mm.blood) controls, and such observation was positively correlated with the mitotic index. In contrast, the spontaneous formation of MN was significantly decreased in the three investigated doses of the extract. In pre- and post-treatment experiments, a similar picture was drawn, and the plant extract was able to modulate the mutagenic effects of MMC.

Abstract
Adherence capability of Pseudomonas aeruginosa isolated fromIraqi patients was tested in accordance to their capability to adhere to epithelial cells. A range of adherence ability was found: Thehighestcapacity with mean number of adhering bacteria of (13.5) was noticed. A sparse content or alginate was detected in these isolates analysis of pilin protein isolated from cultures exposed to ciprofloxacin antibiotic at sub MIC, level using polyacrylamide gel electrophoresis indicated that treatment of cells with 3/4 MIC show highest activity as compared with others. Results of protein profile followed the same trend of that in adhesion experiment which allows to hypothesize that these isolates are piliated, no mucoid.

Abstract
The objective of this study was to assess the effect of licorice extract (LE) on induction of ovulation and on the structural changes of ovary, oviduct and uterus. Divided into 6 groups: G1, G2, G3, G4, G5 and G6 (14 animals for each). G1 received 1g/kg bwt/day of licorice extract for 2 weeks. G2 were fed with 0.5g/kg bwt/day of licorice extract for 2 weeks. Mice fed on normal diet and tap water (without LE) were regarded as control group (G3). G4 and G5 received the same doses of G1 and G2 respectively for 4 weeks. G6 received tap water only and considered as a control for G4 and G5. Before the initiation of dosing, vaginal smears were taken daily and ceased when estrus phase of first ovarian cycle appeared. At the end of the experiment, animals were sacrificed and reproductive system were excised and divided into two sides: the left one only was used for histological studies. The results indicated that LE consumption caused an increase in the diameter of the ovary, total number of follicles and their diameters of all mice treated with a higher dose and longer duration. Dose of 1g/kg bwt/day was also more effective upon measuring the diameters of uterine glands and the diameter of the oviductal ampulla, while the lower dose gave higher results only in measuring the height of endometrial epithelial lining cells and the oviductal epithelial lining cells. There was a significant precocious estrus cycle in animals that treated with 1g/kg bwt/day and a non significant precocity with 0.5g/kg bwt/day, as compared to the control group. These results may be attributed to the effect of various components of LE, especially estrogen-like substances in addition to other components like: proteins, amino acids, vitamins and trace elements present in the licorice. The results of the present study indicate that the use of low doses of LE (0.5, 1)g for a short period of time 2-4 weeks improved the reproductive organs, induced earlier estrus cycle and consequently may improve the reproduction. The consumption of 1g licorice extract daily for 4 weeks induced sexual maturity better than the other dose and period.