Journal of Biotechnology Research Center

Abstract
Calli cultures of stems and leaves explants excised from field -grown rubber, Ficus elastica Decora, plants were formed on agar-solidified Murashige and Skoog (MS) medium.The results proved that MS medium supplemented with 1.0 mg L-1 benzyl adenin (BA) and 0.8mg L-1 2,4-dichloro-phenoxy acetic acid (2,4-D) was suitable to stimulate stem’s callus at ratio 87.5% . Whereas supplementation of MS medium with 0.5 mg L-1 of both BA and Indole -3-butyric acid (IBA)encouraged callus formation to reach76.6%. Leaves showed responses for callus initiation up to 75% at the same media. Stem calli showed limited ability to regenerate shoots on both agar solidified MS medium containing 1.0 mg L-1 2,4-D and kinetin (kin) 0.5 mg L-1 and MS medium supplemented with 0.5 mg L-1 of both BA and IBA. Transferring of shoots to differentiation medium (MS+0.5 mg L-1 BA+ 0.1 mg L-1 IBA) stimulated the growth and elongation of these shoots. Shoots were transferred to agar-solidified MS medium free from growth regulators failed to form roots. Because of the less number of shoots, other rooting media were not tested. The data showed, that shoot tips succeeded to regenerate shoots when they were cultured in different MS. The results proved clear increase in chlorophyll and protein content of the rubber shoots as compared with content of field grown rubber plants. It was noticed that agar-solidified MS medium supplemented with 4.0 mg L-1 BA was considered the optimum medium for shoots regeneration. All plants regenerated from shoot tips were readily rooted in agar-solidified MS medium with increasing Potassium Nitrate KNO3 from 1900 mg L-1 to 2000 mg L-1, and at the same medium supplemented with 3.0 mg L-1 IBA and 1.0 mg L-1 BA. All these plants were successfully acclimated and transferred to peat moss.

Abstract
This study was conducted in plant tissue culture Lab. Horticulture Dept., College of Agriculture, Diyala University to evaluated the effect of plant growth regulators, North and South pole of magnetic field on common mandarin (Citrus reticulata L. Blanco) seedling growth and callus initiation. Seeds were cultured on solidified MS medium to study the impact of increasing magnetic field flood on the production of some plant growth regulators, alkaloids callus initiation. Results can be summarized as follows: Kinetin at concentrations (0.0, 1.0, 2.0, 3.0, 4.0) mg/L for 8 weeks of treatment showed that the control treatment gave taller roots (30) mm while the treatment 4.0 mg/L gave the highest number of shoots 1.7 shoot/ seedling. In the magnetic field experiment, the effect of north and south pole of magnetic field 200 mT for six periods (0 , 2 , 6, 11 , `14 , 18) days, showed that the treatment of 18 days gave the best shoot length 22mm. When the medium supplemented with 2mg/ l 2,4-D enhances callus initiation, while the combine of 2,4-D and TDZ reduced the rate of initiation and growth of callus. The production of hormones and alkaloids from mandarin callus exposed to1, 2 or3 magnets recorded the highest amount of Kin in the presence of 3 magnets giving 125.45 µg/ g while the highest amount of ABA, GA , IAA was found in control callus which gave (91.61, 76.25, 51.18)µg/ g respectively. Alkaloids yield showed the presence of Octapamine in the treatment of 3 magnets yielding 73.74 µg/g. The highest amount of Synephrine reached 366.99 µg/ g in the treatment of 3 magnets while the amount of Tyramine recorded 79.02 µg/ g in the treatment of two magnets. Ephedrine was higher in the callus that not being exposed reached 219.99 µg/ g.

Abstract
Inhibitory activity of Streptococcus thermophilus unconcentrated and concentrated filtrate was studied against some pathogenic bacteria included: Pseudomonas aeruginosa, Klebsiella spp., Staphylococcus aureu, Escherichia coli. Inhibitory activity of protein that extracted from S. thermophilus concentrated filtrate was studied after precipitate by ammonium sulphate and inhibitory activity of lipophilic fraction that extracted from concentrated filtrate with chloroform-methanol (1:1 vol/ vol) was studied against pathogenic bacteria. Also inhibitory activity of biosurfactant produced by S. thermophilus was studied against growth and biofilm formation for pathogenic bacteria. The results showed that unconcentrated and concentrated filtrate had inhibitory activity against all pathogenic bacteria. Also protein and lipophilic fraction had inhibitory effect against pathogenic bacteria, while the biosurfactant show inhibitory activity against growth of all pathogenic bacteria but show inhibitory effect on biofilm formation only for pathogenic bacteria S. aureus and E. coli.

Abstract
Callus cultures were produced from leaf, excised from axenic carnation, Dianthus caryophyllus L. seedling, on agar-solidified MS medium supplemented with 0.5 mgl-1 of 2,4-D and 0.1 mgl-1 of BA. Cell suspension was initiated in liquid MS medium containing the above growth regulators. This suspension reported the density of 143 × 103 cell /ml and its viability was ranged between 41-90%. Samples of cell suspension represents seven different densities were exposed to three electrotreatment 300v/50 msec, 500v/100msec. and 700v/150 msec. These electrotreated samples were cultured by embedding in to agar drops in solid- liquid cultures. Callus derived from cell suspension expressed their totipotency forming carnation plants.

Abstract
This study included the preparation of the mixture of alcoholic extracts of Peganum harmala seeds and Pericarp of Punica granutum at concentration (1+40), (1.5+45), (2+50) mg\ml. To study the influence of the mixture of alcoholic extracts of P. harmala and P. granutum on viability of the protoscolices of Echinococcus granulosus In vitro (In tubes), and In vivo (In albaino white mice injected intraperitoneal with protoscolices). In vitro experiment revealed complete inactivation of protoscolices (death) with concentration (2+ 50) mg/ ml after 30 minute. In vivo a significant reduction in weight of liver and spleen occurred in treated groups in comparison to control (untreated) infected group.

Abstract
The present study revealed that exposure of stem derived callus of Sesamum indicum L. that grown on initiation medium Murashige and Skoog (MS) supplemented with naphthalene acetic acid (NAA 1.0 mg L-1) + benzyl adenine BA 2.0 mg L-1 to 25, 30, 35, 40, 45 or 50◦ C as short term heat shock stHS 5,10 min. and long term heat shock ltHS 15, 20min. enhanced a desirable changes of callus growth. Moreover, the amounts of nucleic acids DNA, RNA, proteins and the specific activity of thymidylate synthase (TS), dihydrofolate reductase (DHFR) and serine hydroxy methyl transeferase (SHMT) enzymes were also increased. The heat shock treatment 40◦ C/15 min. gave the best effect in amount of both DNA and RNA that increased to 95 and 949 µg/g compared to control 58 and 550µg/g callus fresh weight respectively. Total protein was also increased to2.85 mg/g compared to control 1.47 mg/g. Maximum specific activities of TS, DHFR and SHMT enzymes were recorded 3.41, 0.97, 0.31µmol/min/mg protein respectively compared with control 1.70, 0.43, 0.13µmol/min/mg protein. Whereas, heat shock treatments 45 and 50◦ C of both tearms reduced each of callus fresh weight, DNA, RNA amount, and decreased the specific activities of the above enzymes

Abstract
The objective of this study was to investigate the immunological effect of Anastatica hierochuntica extract on albino male mice. The total extract of the raw herb was obtained by using methanol at three different concentrations (25, 50,100)mg/ml. Hydrocortisone used as immunosuppressant drugs (positive control). Adenosine deaminase (ADA) activity, Phagocytosis and immunoglobulin (IgG) were studied with the herbal extracts and appropriate controls. Anastatica hierochuntica extract was significantly increasing IgG level when used at dose 50mg/kg (P≤0.05). It also increases significantly the phagocytosis at dose (50,100) mg/kg after 30min and in dose 50 mg/kg after 60 min (P≤0.05). In addition, there was significant inhibition in ADA activity when used plant extract at dose 100 mg/kg after 2 weeks of treatment. Hydrocortisone was found to lead a statistically significant inhibition of phagocyosis, ADA activity, and decrease the IgG level after 2 weeks of treatment (P≤0.05). In conclusion immunomodulatory activity of Anastatica hierochuntica, related to IgG level, phagocytosis and ADA activity, is revealed during this study. The herbal extract has shown to promote the IgG level, and also the phagocytosis after 2weeks of treatment.

Abstract
The present study aimed to know the inheritance method of elbow crease trait in human from parents to their offspring, our study included (100) families (parents, sons and daughters) however we have studied the effect of parent degree of consanguinity, blood groups on the transmission of this trait from parents to their offspring. The result indicates there was no significant association between parent’s degree of consanguinity and transmission of elbow crease because the ratio of randomly selected foreigner parents were %65. In addition our study involved the study of some other factors like blood groups which has also no significant effect. There were several significant association of presence of elbow crease in both parents and their offspring.

Abstract
Overuse of antibiotics has become the major factor for the emergence and dissemination of multi-drug resistant strains of several groups of microorganisms and this lead to search for agents that may have antibacterial effects. Vitamin E emerged as an essential, fat-soluble nutrient in the human body and it is essential, because the body cannot manufacture its own vitamin E, so foods and supplements must provide it. The aim of the present study was to evaluate the effect of vitamin E against pathogenic bacteria. Gram positive and gram negative bacteria were selected as the test microorganisms based on their importance in infections. In this study vitamin E used in four concentrations (50,100,200,400) IU/ml. The agar diffusion method was used to determine antibacterial activity. Results showed that gram negative bacteria were shown to be more resistant than gram positive bacteria. The resistance of gram negative bacteria towards antibacterial substances may be related to lipopolysaccharides in their outer membrane.

Abstract
A total of one hundred five samples were collected from hospitals of Baghdad city during the period from 10/12/2008 to 15/3/2009. These clinical samples included: urine (50) wound swabs (25), sputum (20), and ear swabs (10). These samples were collected from the Baghdad hospital/ Teaching Laboratories, and Al-Yarmook Hospital/ Teaching Laboratories. Twenty six isolates of Proteus mirabilis were characterized according to the morphology and microscopic characteristics, along with the biochemical and confirmatory APi 20 E tests. These isolates were obtained from: urine (19), wound swabs (6), ear swabs (2), and sputum (1). The twenty selected isolates were tested for resistance against ten antibiotics and only urine samples were tested for nalidixic acid and nitrofurantoin resistance. It was shown that there were differences in the antibiotic resistance of isolates. High resistance to nitrofurantoin and ampicillin were found among isolates as (100%) and (75%) respectively while the resistance of Proteus isolates to trimethoprin /sulphamethazol, were (65%). This study also showed that resistance of isolates to gentamicin, ciprofloxacin, ceftazidime, pipracillin, cefotaxime, nalidixic acid azteronam, imipenem and amikacin were (50, 40, 40, 40, 35,27, 20,15, 5)% respectively.

Abstract
This project was conducted to study the activity of secondary metabolites produced by one of a human microflora which is Lactococcus lactis on AMN-3 tumour cell line in vitro. Twenty samples of dairy products (bucolic sour yoghurts, pasteurized milk and raw milk) were collected; nine isolates of Lactococccus were isolated by propagating in MRS broth medium followed by subjecting the isolates to microscopic, cultural, physiological and biochemical tests. The results showed the presence of four isolates belonged to the genus Lactococcus sp. lactis. the isolates were grown in three broth production media, which were MRS, VVM and BHI at 37ºC for 6 h. followed by extraction of crude extracts using ammonium sulphate saturation ratio which was 80% for each medium confidentially. The overall result was three crude extracts (one from each medium), followed by estimation the concentrations of extracted proteins depending on standard curve of bovine serum albumin. The cytotoxic activity of different concentrations for each crude extract was studied on AMN-3 tumour cell line for three incubation periods (24,48,72) hrs in addition to normal rabbit embryo fibroblast (REF) cell line for 72 hr only. The result illustrated a clear cytotoxic activity of these crude extracts with high significances on this tumour cell line during the three incubation periods, suggesting that the cytotoxic effect of these crude extracts is a dose and time dependent, but on REF cell line, there is no significant effect of these crude extracts was reported, suggesting also, that may be the active compound of L. lactis possess some specificity in cytotoxicity on cancer cells but not on normal cells.

The locally isolated Pseudomonas aeruginosa NW was tested for their ability to produce alcohol dehydrogenase (ADH) enzyme by growing the bacteria on three different aromatic hydrocarbons: naphthalene, xylene and toluene. Following five days incubation the bacterium showed a good ability to utilize xylene, limited ability for toluene utilization and failed to degrade naphthalene. The ADH activity was assayed following each day of incubation on xylene as sole source of carbon and energy and it was found that a maximum ADH volume activity was 2.86 and 4.11 U/ml following the third and fourth day of incubation, respectively. The production of ADH was optimized under different conditions involving: temperature, pH, xylene concentration and incubation period. Results indicated that the optimal conditions for ADH production were by incubating the bacterium in MSM medium containing 1% xylene, pH: 8 for four days and incubation at 37ºC.

Abstract
Study the resistance of bacterial and fungal isolates which identified previously towards different antibiotics that have been recommended for treating those patients. It was found that higher percentage of resistance to penicillin, gentamycin, tetracyclin, chlormphenicol, and trimethoprim- sulfmethazxol then moderate resistance to ampcillin, nalidixic acid, cephotaxime, streptomycin, cephalexin, rifampicin and less resistance to ciprofloxacin that represent the most effective antibiotic against uropathogenic bacteria. Ketazole antibiotic was used against yeasts, which the minimum inhibitory concentration of ketazole against 83% of Candida. albicans isolates was 128 μg/ml, the rest of the isolates was 64 μg/ml. Isolation of bacterial and yeasts DNA (chromosome + plasmids), results of DNA isolation showed that bacterial isolates (E.coli (RS1), proteus. mirabilis (RS2), klebsiella. pneumonia (RS3) and Pseudomonas. aeruginosa (RS4) have chromosome and plasmids vary in numbers and size according to isolate. While yeast (C13) isolate have only chromosome without plasmid.
A result of curring shows that:
-Resistance to ampicillin, tetracycline, gentamycin, pencillin for all bacterial isolates carried on plasmid.
- Resistance to cephalexin by RS1 isolate, resistance to cephotaxim by RS3 isolate and resistance to chlormphnicol by RS4 isolate carried on plasmid, while resistance to other antibiotics were carried on chromosome.

Abstract:
This study was designed to test the lipid-lowering and antidiabetic activities of olive leaf and its callus extract. Diabetes in mice was induced by intraperitoneal injections of alloxan. The serum glucose and serum lipid were examined. Diabetic mice showed hypeglycemia, hypelipidemia. The administration, for 2 weeks of olive leaf and its callus extracts significantly decreased the Total cholesterol (TC). Triglycerides (TG). Low density lipoprotein (LDL), very low density lipoprotein (VLDL). Both types of olive extracts had significant hypoglycemic effects on blood glucose levels in diabetic mice. This hypoglycemic effect was as potent as the hypoglycemic effect of insulin. However, the callus extract was more potent than the leaves extracts and most potent than insulin in causing a significant decrease in LDL, VLDL, TC, TG and in antidiabetic effects.