Journal of Biotechnology Research Center

Abstract
Pot factorial experiment was conducted,using Completely Randomized Design (CRD) with silty loam soil in winter season of 2011-2012 in lath – house at the College of Agriculture University of AL-Muthanna to study the effect of mycorhiza fungus Glomus mosseae in phosphorus availability and growth of wheat plant. Three levels used of rock phosphate (0 , 60 , 120 kg P.hr-1 ) and inoculation of soil, and without inoculation by mycorrhiza by three replication. Results showed that the application of rock phosphate with mycorrhiza fungus caused significant increasing in height of the plant, shoot dry matter, root dry matter and available phosphorus in soil to two growth period where increase percent which was 22.16 , 16.94% to height of plant and 28.68 , 58.46% to shoot dry matter and 24.39, 34.26% to available phosphorus in soil to first and second growth period succession compared with control treatment.

Abstract
The study was conducted to evaluate the inhibition activity of alcohol extract of Peganum harmala seeds in some pathogenic fungi (Aspergillus niger, Aspergillus flavus and Penicillium spp.), which was isolated in food research center/ Ministry of Science and Technology. Results revealed that alcohol extract of Peganum harmala seeds was inhibited the growth of all kind of fungi under study at the extract concentrations (0.012,0.05, 0.25,0.5,1,5,20,50, 100 ,200 and 300 mg/ml), colonies diameters for Aspergillus niger was 27, 25 mm at the concentrations 0.012 and 0.05 mg/ml respectively in the comparison with its colonies diameter in the control group which was 38 mm for both concentrations, and it was 26,21 mm for Aspergillus flavus at the same concentration (0.012 and 0.05 mg/ml) respectively, whereas the colonies diameter was decreased to 9,7 mm for Penicillium spp. at 0.012 and 0.05 mg/ml of alcohol extract respectively. In general, the inhibition activity of Peganum harmala seeds alcohol extract was increased with the increasing of extract concentrations. The growth rate of all kinds of fungi under study was completely inhibited at 0.25mg/ml concentration and above.

Abstract
Three hybrids of American cucumber were studied, they are Lot number EM285 labeled number 1, Hybrid tocaeen F1 labeled number 2, Hybrid super green F1 labeled number 3. The seeds of these hybrids were exposure to four frequencies of electric current 0,1,2 and 2.5 Am for 5 minutes and 4 periods of ultra violate rays UV at 254 nm. For 0,30,60 and 90 second. The sterilized seeds were cultured in MS medium. After 3 weeks apical meristems of seedling were excise and cultured in new MS medium and of 5 weeks shoot systems were taken to study the element content nitrogen, phosphorous, potassium, calcium and magnesium in addition to estimate protein percentage. The results showed that hybrid 3 had a significant increase of element contents and protein percentage compared with hybrid1 and 2.The interactions between hybrid3 and 90 sec. UV exposure showed the highest content of calcium element reached 2.73 mg.gm-1 with an increase of 184.37%compared with the control. The interaction between 2.5 Am and 90 sec. UV exposure showed the highest rate of protein reached 15.5%with significant differences of all interactions.

Abstract
The phenolic compound and other active compounds were extracted from cinnamon, in an attempt to produce a drug from Medical and herbal Plants for the diabetes on rats. The extraction process included two methods the first carrid out with 2% v/v acetic acid and propanol, and in the second process 70% v/v ethanol and chloroform were used. In the detection of extracts it was noticed that both conteined phenols, flavonoids, tannins and Glycosides and Coumarins, while the saponin showed up only in the alcoholic extract of cinnamon. The phenolic compounds in the extracts of cinnamon were determined by using Ultraviolet Spectroscopy (UV) and other function groups such as phenols, aldehyedes, estars, hydrocarbons, alcohols and carboxylic compounds were diagnosed by using Infrared spectroscopy (IR). The study of the half lethal dose (midl lethal) (LD50) was also examined by testing the toxicity of the two cinnamon extracts. Rats were used as laboratory animals in conducting the experiments. The rats were fed by the dose (100, 200, 300,400, 500 mg / kg of body weight). The results have shown that there is no toxic effect in the cinnamon extracts. Similar concentrations were used to determine the effective dose of the extracts in reducing the level of glucose in the blood of healthy animals. The dose (300) mg / kg of body weight of the animal body has proved to be the the most efficient as it decreased. The fasting glucose (Glu) serum consider by following of the cinnamon extracts with the adopted groups during the duration of (30) days in comparison with the control group (the infected) and outperformed the treated given Glibenelinide (daonil) group. The biochemical tests showed that the serum concentrations of each of cholesterol (TC),Triglyceride (TG), low density lipoprotein (LDL-C) was lowered in each of the groups treated with both cinnamon extracts in comparison with the positive groups (infected) throughout the duration (30) days and overcome the daonil treated group. However the serum high density lipoprotein (HDL-C) level was increased in cinnamon extract treated groups and decreased in the control group (infected).
The serum concentrations of enzymes (GOT) and (GPT) were measured to test the liver function after where their levels were lowered after (30) days in the cinnamon extracts administered groups compared with the two other groups, the control (positive) and the Daonil. The Urea and Creatinine levels were measured to determine or test the kidney functions, where it was observed as from the early days of the infection in that animals infected by induced diabetes developed symptoms of the illness, apparent in the high levels of urine which affected the functions of kidney, but the long duration (30 days) of treatment with these extracts of cinnamon proved their efficiency over the control group (positive).

Abstract
This study was carried out on fruit of Hillawi date palm cultivar which pollinated by Khukri Adi and Ghunami Akhder cultivars to study the effect of NAA 25 ppm and pollen extraction of Khukri Adi and Ghunami Akhder 2g/L which were sprayed at 30 and 45 day after pollination on gene expression by studying the changes in protein pattern during fruit development and ripening by gel electrophoreses method with sodium dodecyle sulfate (SDS-PAGE). The result showed there were many protein band which were different in molecular weight and numbers on the gel with changes on fruit growth stages and treatments, These bands of protein were similar in early stage of growth but different in the period of 13,15 weeks after pollination, this difference may be correlated with ripening date. Our results showed that we could consider the protein pattern as gene expression markers for fruit and linking them with a different growth and ripening stage and explanation of metaxenia phenomenon

Abstract
Three nitrogen fixing Cyanobacteria purified isolates were selected which express the three morphological states, first Oscillatoria chlorina as an example of filamentous non Heterocysts Cyanobacteria and second Microcystis rubusta as an example of Unicellular Cyanobactera and third Anabaena spiroides as an example of filamentous Cyanobacteria containing a Hetrocysis, these isolate were cultured on ASM-1 free nitrogen medium using light tension of lux (2500) and temperature of 25°C. Total DNA were isolated and found that its concentration varied between (1.3 to 1.8) the concentration was fixed in order to obtain that each 100 l contains 50 ng/l. Primers for RAPD-PCR were used to detect the genetic variation between the three genera of Cyanobacteria understudy and in order to diagnose the genetic relationship and to detect the genetic dimension between these isolates in order to use these results to found the genetic fingerprint. Eleven primers were used in RAPD-PCR reaction and 10 of them were positive. The OPC-05 gave the high number of bands (20 bands) while OPA-18 and OPJ-13 gave the lowest (9 bands).The genetic dimension between the three isolates were compatible with the morphological, microscopical and physiological results obtained as we found that Microcyctis rubusta and Anabaena spiroides were closely related and Oscillatoria chlorine was far genetically.

Abstract
Eggs, larvae, pupa and adults of date moth Ephestia cautella were exposed to three levels of microwave power 600, 800 and 1000 W at different exposure time (0,6,8,10,12,14,16,18,20 and 20 second). The results showed that microwave had high ability to kill different stages of fig moth. The effect increased by increasing of microwaves power and exposure time. The percentage of egg mortality was %100 at 600 and 800 W, and exposure time. Similarly, egg mortality was also increased at 1000 W in less than 12 sec. The highest motility of larvae was %90 at 600 W and %96.67 at 800 W, when the exposure time was 10 sec., while, the mortality was %100 at 1000 W and 18 sec. male and female pupal mortality rates was %100 at 800 W, when the exposure times were 20 and 18 sec., respectively. The ranges of lethal time of %50 and %95 ( LT 50 , LT 95 ) at 600-1000 W were 501-9.9 and 15.7 – 34.2 , 3.0-7.5 and 9.0-50.0and 3.2-4.8 and 7.9-39.1 for egg , larvae and pupae, respectively. However, the results showed that all adult treatments significantly affected age average and emergence of adult male and female. The age of emerged males and females did not exceeded 4.267 and 5.267 days respectively, at 600 W and 6 sec. exposure. While, adult emergence stopped at 800 W and 14 and 16 sec. for male and female pupae respectively. The results of this investigation indicated the affectivity of using microwaves technique and further research efforts is needed before the feasibility microwave technique can be evaluated as alternative in stored dates chemical method using Methyl Bromide

Abstract
This study succeeded in establishment carrot cell suspension cultures from stem callus in liquid MS medium containing 1.0 mg L-1 of each NAA and BA. Its density approach 3.4 ×105 cell ml-1 at the third day of culture. These cells ,continued, when embedded in agar drop using Multiple Drop Array (MDA) technique, in division and forming cellular colonies which producing numerous callus primordia that developed to callus cultures. When transferred to the solid differentiation medium (MS+ 1.0 mg L-1 NAA+ 1.5 mg L 1 BA), two hundreds and seventy-three shoots produced. They readily rooted in agar-solidified MSO medium after three weeks and adapted in conditions of culture room. They were not transferred to field due to the unfavorable environmental conditions at that time. Cell suspension-derived callus tissues contained 82.62 μg anthocyanin gm-1 of callus fresh weight, and 0.555 mg beta-carotene 100 gm-1 of callus fresh weight in the third period of extraction compared with their quantity calculated in stems callus which recorded 52.4 μg gm-1 and 1.988 mg gm-1 respectively.

Abstract
The research was conducted to study the effect of adding different concentrations of Salicylic acid (0,50,100,150,200) mg/ l in the production of some secondary metabolic compounds of plant(garlic) Allium sativum L. Quantitative and qualitative analysis of secondary metabolites were estimated by using( HPLC ). Garlic cloves were culture on MS media supplemented with 2mg/l of 2,4-D, 0.5mg/l of NAA and 1mg/l of BA for callus induction. Adding salicylic acid (100 mg/l) cause in significant increase of fresh weight 734mg and volatile oil in callus extract compared.

Abstract
This study was done to test the activity of watery and alcohol extract of the fruit of Berberis vulagaris against of two species of gram positive bacteria Bacillus cereus and Staphylococcus aureus also ,two species of negative gram bacteria E. coli and Pseudomonas aeruginosa by agar well diffusion method, the results showed that the alcohol extract of the Berberis vulagaris with high activity against gram positive and the effect was increase with the increasing of the concentration. While the watery and alcohol extract has a high effect on gram negative bacteria which effect only with high concentration, while there is no effect in low concentration and the results showed that the alcohol extract of the Berberis vulagaris with high activity against gram positive compare with watery extract. MIC and MBC were measured and the results were identical for the previous results, for the effect of both extracts on the growth of gram negative and positive bacteria HPLC analysis was don for Berberis vulagaris which is quantitative and qualitative assay to measure the concentration of some alkaloid compound Tetrandrine, Chondocurine, bamine, which be with high ratio on Tetrandrine compound 38.42, the rate of alkaloid compound in fruit of Berberis vulagaris was higher .

Abstract
Two field experiment were conducted at farm of Field Crops Dept. Coll. of Agaric, Univ of Baghdad during winter seasons of 2010/2011, 2011/2012 to study effect of spraying of growth regulators Benzyl adenine (BA), Zinc and Boron separately or combined during important stages of plant faba beans (Spanish var.) on yield faba bean and its components. A split- split plot with R.C.B.D. distribution was used by three replications. Levels of Benzyl adenine 0, 75 , 150 mg.l-1 (BA0 , BA1, BA2) respectively occupied the main plots. Zinc treatment 0,25,50mg.l-1 (Zn0 , Zn1 , Zn2) respectively occupied the sub-plots. While Boron treatment 0, 150, 300, 450 mg.l-1 (B0 , B1 , B2, B3) respectively occupied the sub-sub plots. The results showed that spraying plants with Benzyl adenine at concentration of BA2 (150 mg.l-1) or Zinc concentration of zn2 (50 mg.l-1) or Boron at concentration of B3 (450 mg.l-1) increased significantly all studied characteristics of both seasons. They give the highest fertility rate, number of pods per plant, number of seeds per pod and total seed yield. While 100 seed weight was increased significantly with using BA1 (75 mg.l-1) or zinc at Zn2 (50 mg.l-1) or boron at B2 (300 mg.l-1) as compared to the control treatment Results revealed that interaction between the plant treated with 150 mg.l-1 (BA) concentration, zinc with 50 mg.l-1 and boron with 450 mg.l-1 increased significantly all the studied characteristics and seed by 44% in of both seasons.

Abstract
This study was set to evaluate the testosterone serum levels and their association with insulin resistance to uncover if any related possible negative consequences in Iraqi patients with Diabetes mellitus (DM). Sixty six men with type 2 DM were randomly selected and compared to 18 healthy volunteers, demographic data were collected in addition to 10 ml of venous blood was drawn to evaluate serum levels of total Testosterone (T), free testosterone (FT), c-peptide, Sex Hormone Binding Globulin (SHBG), lipid profile, and blood sugar. The mean serum level of total testosterone was 2.434 ± 0.12 ng/ml in diabetic patients and 3.62 ± 0.32 ng/ml in non-diabetic subjects which was significantly different (p=0.003). The mean value of free serum level of testosterone was significantly higher in non-diabetic group (14.71 ± 1.4 pg/ml) in comparison with diabetic group (10.71 ± 0.54 pg/ml; p=0.017). Total testosterone correlates positively with FT and SHBG, while negatively with c-peptide, age and Body mass index (BMI). It’s concluded that total testosterone, free testosterone, and SHBG were significantly lower in diabetic group which strongly associated with insulin resistance.

Abstract
A new immunoblot assay, composed of five Epstein-Barr virus (EBV)-encoded recombinant proteins virus capsid antigen [VCA] gp125, p19, p22, early antigen [EA], and EBNA-1 IgG, was used to manifest the EBV infection and look at the antibody pattern to EBV proteins in the serum of both autoimmune disorders and Thalassemia patients and compare the observations with those in normal healthy controls. Serum samples from 35 rheumatoid arthritis patients, 20 SLE, 20 autoimmune hypothyroid diseases, 35 Thalassemia patients and 20 healthy controls were tested for EBV IgG antibodies by an immunoblot assay (Euroline). The results showed that the high percentage recorded was 50% in acute infection. Followed by 30% at late infection, while late phase with loss EBNA-1 and reactivated infection were 10% compared to the normal healthy controls. Our study showed an increased EBV activation among the autoimmune patient groups compared to the normal healthy controls. Further studies are required to delineate the association between the etiology of autoimmune disorders and EBV.

Abstract
During the period from September 2007 till December 2007, a total of two hundred samples were collected randomly from children patient attended to Al-Mansour General Hospital for Pediatric in Baghdad to determine the prevalence of intestinal protozoa (E. histolytica and G. lamblia) among patients with diarrhea or abdominal symptoms referred from the out patients. General stool examination was preformed for all individuals included in the study. The direct smear method by normal saline 0.9% Nacl and iodine-lugol solution staining method was used. Also complete blood picture test was preformed for all patients to evaluate, PCV, total white blood cell count, differential white blood cell count. It was found that the total prevalence rate of (E.histolytica and G. lamblia) infection was (34.5%) of which E. histolytica patients was (27.5%) and G. lamblia patients was 7%E. histolytica was significantly more common (45%) in school age children (6-10) years, while G. lamblia was more frequent 53.3% in preschool age 1-5 years. Mean of haemoglobuline level 10.36 for E. histolytica, while mean of haemoglobuline level 9.73 for G. lamblia. As a result, infections with intestinal protozoa dose not lead to anemia in great percentage, but in low percentage. Bloody stool was associated with positive stool for E. histolytica trophozoites and cyst, while isolation of G. lamblia had no association with bloody stool.

Abstract
This study was carried out in order to determine the toxic, mutagenic and anti-mutageniceffects for aqueous extract of Vincaroseadried leaves against the mutagenic effect of Methotrexate (MTX) as a chemical mutagen and Ultra Violet rays (UV) as a physical mutagen. The effect was studied in a bacterial system (G-system). The used system consisted of three wild isolates G3Bacillus spp., G12Arthrobacter spp.and G27Brevibacterium spp., The studydepended on recording survival fraction (Sx) as an indicator for the induction of Streptomycin and Rifampicin resistance mutants as a genetic marker.Aqueous extract was prepared from dried leaves, gradual concentrations of plant aqueous extract was used to choose the suitable concentration which is remembered the negative control, the optimum concentration of aqueous extract was 200 µl/ml comparing with the negative control. The interactions included three types of treatments (pre-mutagen, with-mutagen and post-mutagen) in order to determine the mechanisms of this plant extracts in preventing or reducing the genotoxic effect of mutagen (MTX and UV).The results showed that the interaction effect between the optimum concentration of aqueous extract and the mutagen on survival fraction (Sx) increase the survival fraction value of the of G-system isolates to reach normal value compare with positive control (MTX), the results of the interaction between optimum concentration for extracts and the treatment with mutagen to induce resistance mutant for streptomycin and rifampicin found that the MTX had no effect to induce resistance mutant for these two antibiotics, for all types of treatment (pre-MTX, with-MTX, and post-MTX), the aqueous extract suppress or repair mutant and give 100% protection for bacterial cells, while the UV had some effects to induce resistance mutant for these two antibiotics for pre-UV and within-UV treatments and the aqueous extract showed suppressing or repairing mutant partially with post-UV treatment and gave 100% protection for bacterial cells.

Abstract
This study was reflected on the relationship between the polycystic ovary syndrome (PCOS) and the genetic alternations in TPO gene. Fifty infertile Iraqi women with PCOS and 20 healthy women were included in this study, Blood samples were collected from the Infertility center of AL-Yarmok Teaching Hospital in Baghdad, during the period from November, 2010 to May, 2011. The age of infertile and fertile women was ranged from 16 to 45 years. The results of hormonal assay were as follows: There is significant (P ≤0.05) decrease in E2 and FSH levels in PCOS women and fertile women, There is significant (P ≤0.05) increase in LH levels in PCOS women and fertile women. There is no significant differences in Testosterone levels and the ratio of LH/FSH was ≥1.5. The molecular study was focused on the 18% of PCOS women with hypothyroidism. By sequencing for 27 samples; two novel different mutations were identified in the reading frame of the TPO gene in transcript variant of exon 9: c.1471delC (deletion C in codon 460) and c.1481delC (deletion C in codon 464). The percentage of mutations c.1481delC and c.1471delC recorded 55% and 44% of PCOS with hypothyroidism; respectively.

Abstract
Human papilloma virus (HPV) infection is a causative factor for cervical cancer. Early detection of high risk HPV types might help to identify women at high risk of cervical cancer. The aim of the present study was to determine the occurrence ofhigh risk HPV infection in population of Iraqi women in Baghdad by using Multiplex PCR determine the percentage and genotyping of Human Papilloma Virus and to put the best prevention and control program in Iraqi women. Study started at January 2009 to March 2010, cervical samples were collected from 856 women aged 16–70. HPV DNA amplification was performed using HPV High Risk Typing PCR Kit test for qualitative detection and genotyping of HPV types 16, 18, 31, 33, 35, 39, 45, 52, 56, 58, 59, 66 in the cervical swabs. HPV was detected in 106 ( 12,38% ) of the study population, with a range of 16-70 years age groups. Results showed that the overall HPV prevalence twelve genotypes were identified, including HPV-33 (18.60%), HPV-35 (18.60%), HPV-56 (18.60%) ,HPV-39(10.85%), HPV-52 (10.08%), HPV-18 (7.75%), HPV-16 (4.65%), HPV-59 (4.65%), HPV-58(2.32%), HPV-31 (1.55%), HPV-45(1.55% ) and HPV-66( 0.77%). Of 856, 218 women was also tested by pap smear , with normal cytology was 198 ( 90.83%), 24(12.12%) of them were HPV positive, those with abnormal cytology was 20 (9.17 %), 5( 25%)of them was HPV positive. In this study unlike other epidemiological studies, HPV33,35,56 was the most frequent type (55.8%) in Baghdad, followed by HPV39, HPV52, HPV18, HPV16

Abstract:
The Down syndrome (DS) is the well-known trisomy, which is caused by additional copy of chromosome 21.There are three types of DS. First, fully trisomy (47,XY,+21or 47,XX,+21). Second, translocation DS which result as translocation between chromosome 14 and 21(46,XY,trans (14:21) or (46,XX,trans (14:21)). And the third type the mosaic DS that two cell lines present in the individual. Mostly, studies indicate that frequency of each type 95%, 4% and 1% respectively. Our study aims to estimate the frequency of each of the three types of DS chromosomal abnormalities in Iraqi samples. Chromosomal analysis using G-band technique was performed for 200 Down syndrome cases and 168 of their parents (whenever there were mosaic DS cases their parents were submitted chromosomal analysis). Fifty-seven percentage of cases were fully Trisomy, forty-three percentage of them were mosaic DS, and no translocation pattern was recorded. The maternal ages were between 25-45 for the mosaic DS mothers. According to this study, the frequency of mosaic DS was varied. It seems more investigations need to be done for larger number of DS, and the impact of environmental changes in last decades need to be studied more to be sure of its role in increasing of the proportion of this type of Down syndrome.

Abstract
Breast cancer is the most common malignant tumor and the leading cause of carcinoma death in women. The tumor microenvironment consist from cellular microenvironment, cytokines and chemokines. Its plays an important role in tumor development such as initiation, progression, metastasis and drug resistance. It is becoming increasingly apparent that the malignant tumor progression is maintained by dynamic interplay between tumor cells and many distinct cell types existing in the adjacent microenvironment. Objective of this study is to detect of TNF-α in tumor microenvironment in histological sections of breast cancer and compare it with that of benign breast lesions. This study included 32 patients with breast cancer and 21 patients with benign breast lesions, prognostic factors were registered including: age, histopathological subtype, degree of differentiation& stage of breast carcinoma. Results revealed positive expression of TNF-α in 20 cases, while 12 cases were negative out of 32 samples of breast cancer. In benign lesions 11 cases were positive, While 10cases were negative. Statistical analysis showed significant difference in rate of TNF-α expression between malignant breast samples and benign samples(P ≤0.05).