Journal of Biotechnology Research Center

Abstract
Experiment carried out in the fenced area of botanic garden in the Department of Life Sciences, Faculty of Education Ibn al-Haytham, Baghdad University. During the growing season of 2013-2014 to determine the NPKZn fertilizer and spraying of four concentrations of Gibberellin (25,50,100 and 150) mg.l -1 and the interaction between them in addition to the control treatment in the quality and rheological characteristics of three varieties of the Iraqi wheat (Bhooth 22,AL-fatih and bhooth 128). The layout of the experiment was (R C B D) with three replications. Results showed significant differences between the fertilizer application in all studied characters, and that spraying with 100 mg. liter -1 Gibberellin and gave the highest averages for quality and rheological characteristics except flour recipe portability absorption of water, the interaction between the level of 160 kg.h-1 of manure and spraying Gibberellin significant differences for each of the characteristics compared with the control treatment except for ashes ratio of coarse bran.

Abstract
The aim of this study was to determine the optimum conditions for L-asparaginase production form local isolate Erwinia caratovora AH88 (gained from previous study) by using submerged culture. The results of this study revealed that the optimum conditions for L-asparaginase production from this isolate were include using1 % of lactose as carbon source, 1% yeast extract as nitrogen source, in addition to asparagine with 0.19 % as inducer. The optimum pH was 7, optimum temperature was 30 Co, aeration speed was 150 rpm, optimum size of inoculum was 2% (contain 3 X 106 CFU/ ml) and incubation period was 24 hours. Under these optimum conditions for L- asparaginase production the results revealed that the enzyme activity and specific activity were 71.0 unit / ml and 17 unit / mg respectively.

Abstract
Infertility is a global health problem and one of the major problems in many countries, including Middle East, this phenomenon grow in recent years, as the world has seen a rise in the rates of infertility, especially women infertility, so this study conducted to find some reasons of unexplained women infertility. The current study included (100) infertile women in Anbar province, the ages ranged from 20-45 years of revisions to Al-Ramadi Education Hospital for women and pediatric and some medical clinics in a number of cities and districts of Anbar province, through the period from September / 2013 to January / 2014, in addition to (40) healthy women who did not have any history or clinical signs of infertility and their ages matches of the study group, the results of the current study showed that the proportion of primary infertility in Anbar province, the highest of secondary infertility, amounting to 60% and 40% respectively. The study also included some hormonal tests to members of study and control samples included measurement of Luteal hormone concentration (LH), Follicle Stimulation hormone (FSH), Prolactin hormone (PRL) and Testosterone Hormone (Testo) to excluding any sample suffer from hormonal disorders the study showed a significant increase P≤0.05 in the concentrations of infertile women hormones as compared with the control group, and it was found among the (100) infertile women that 59 (59%)women have different hormonal disorders. A molecular study of 28 infertile woman (do not suffer from any hormonal disorders) in addition to (30) healthy women from the control samples for the detection of polymorphism and disorders that may occur to the aromatase gene CYP19A1 using a pair of specific primers designed throughout this study to determining its relationship with unexplained infertility in women, and showing that there disorders in 25 (89.3%) sample of infertile women ranged from the absence of the primers bands or appearance differs from the expected size while the control group did not show any disturbance.

Abstract
Fusarium wilt disease one of the most important disease worldwide. This study was carried out to evaluate the effectiveness of cabbage leaves for three concentrations of (2.5, 5 and 10g/ 1kg) in controlling soil fungus Fusarium oxysporum that causes wilting pepper. The results showed that the effectiveness of fresh cabbage leaves in reducing the severity of infection and the percentage of pepper wilt disease with significant differences in comparison with the control treatment. The cabbage leaves added to the soil at concentration of 10 g / 1 kg soil achieved the highest percentage of reduction in the severity of the disease and in at percentage of the disease, amounting to 8.75% and 0.5%, respectively, followed by Cabbage leaves treatment concentration of 5 g / 1 kg soil at 35% and 5%, respectively then followed by treatment with Cabbage leaves a concentration of 2.5 g / 1 kg soil at 55%, 10%, respectively, which all differed significantly from the treatment with pathogenic fungus F. oxysporum showed that the severity of the infection and the percentage of infection were 82% 0.45%, respectively. The results also showed that all Cabbage leaves treatment with concentrations of 2.5, 5 and 10 g / 1 kg soil has not adversely affected the growth standards parameters for Pepper plants length, width of their leaves, and fresh and dry weight of the plants after 40 days of cultivation of pepper seedlings as it did not differ significantly from the control treatment. While the results showed that there were significant differences in the value of vigor index for treatments with Cabbage leaves which ranged from 2663 - 2745 compared to the treatment of pathogenic fungus F. oxysporum, which amounted to 1545 while not significantly differ from the treatment of the control as a vigor index reached in 2574.

Abstract
This study was conducted to evaluate the efficacy of biological control agents the fungus Trichoderma viridi (Tv) and the bacteria pseudomonas fluorescens (Pf) against two pathogenic fungi Rhizoctonia solani and Macrophomina phaseolina in Sesame plants under the laboratory and the glass house conditions. The pathogenicity test of both pathogen fungi isolates on sesame seedlings were as follow, the Rs-1 for R.solani gave incidence percentage of 100% and 80% and isolate Mp-8 for M.phaseolina gave 100% and 90% infection under laboratory and green house conditions, respectively. In antagonistic test the Tv-4 isolate gave antagonistic degree 1.2 and 1.3 against the isolate Rs-1 and Mp-8 , respectively . The Pf-6 isolate caused complete inhibition of both pathogen fungi .The study showed superiority of both bio control agents in reducing of incidence percentage from 80% in infested control to 40 and 45 % in Pf and Tv treatments , respectively , after 60 days of planting . These results caused highly increasing in number of pods (3.5 pods / plant ) for each of both bio control agents compared with infested control ( 1.8 pods / plant ). Bio control agents treatments has led to reduce the density of the fungi nursing in their treatment.

Abstract
75 isolates of Pseudomonas aeruginosa was obtained from different clinical conditions. The result showed that 54 isolates which were related to P. aeruginosa have tox A gene will a ratio (72%), while 21 isolates will a ratio (28%) from the isolates didn’t have this gene and when we compared the duplicated bands with a suitable DNA marker, the size of the gene was found to be 352 base pair. The analysis of nucleotide sequence for tox A genes of P. aeruginosa was revealed there was some mutations in the DNA of this gene occurred in the nitrogen base sequence which almost included substitution type and insertion. The percentage of identities 96 and 99 % with the original gene. DNA sequencing analysis of amino acid translation of tox A genes revealed that most isolates display different point mutation as compared with NCBI data. Point mutation was detection in P. aeruginosa tox A causes conversion of Alanine to Threonine, Alanine to Glutamine, Alanine to Glycine, Leucine to Valine and Arginine to Leucine.

Abstract
This study was conducted on a total of 67crossbred cows in Erbil plain during the period from July/ 2014 to April / 2015, to study the role of sex chromatin shapes (Drum stick, Sessile nodule, Tear drop and Small club) on Daily Milk Yield (DMY), Lactation Period(LP), fertility rate and fertilize estrous sequences. We also study the effect of age of cows on the DMY, LP, and fertility and fertilize estrous sequences. The overall means of sex chromatin shapes drum stick, sessile nodule, tear drop shape and small club were 25.0 %, 35.0 %, 22.0 % and 18.0%, respectively. While the overall means of (DMY), LP, fertility rate and fertilize estrous sequences were 23.99kg, 197.46 days, and 87.13% and 1.54, respectively. The results showed that there is a significant (p≤0.05) different in relation to DMY, the highest rate of DMY showed among the cow, arise sex chromatin drum stick (28.86kg), lactation period (203.57), the small rate of DMY was low rate among the cows which arise sex chromatin tear drop and LP (192.63days). Significant effect of ages of cows on the patterns of sex chromatin had been observed. It was concluded from this study that the prediction of production milk production and reproduction (fertility rate and fertilize estrous sequences) performance of crossbred cows from shapes and percentage of sex chromatin distribution and measurements. The sex chromatin shapes in doing early selection the elite individuals on the linkage between sex chromatin shapes and performance production

Abstract
Breast cancer is a heterogeneous disease and remains one of the most leading causes of death among women world wild. Anti-apoptotic Bcl-2 family members inhibits cell death and promote proliferation of cancer cells. Overexpression of Bcl-2 has been shown to inhibit the initiation of apoptosis, in the presence of some stimuli including anticancer drugs in a number of systems. To study the expression of Bcl-2 tissue microarray (TMA) investigation was designed and constructed. Using 3 TMAs; the second for the 50 Iraqi breast cancer cases, one for the 30 Italian cases and the third for the 10 benign cases. Each sample represented in a triplicate in the recipient block, all these TMAs expressed to the same condition for construction and IHC staining. TMAs are a powerful, fast and economic technique. IHC was used to study the Bcl-2 expression. The results are represented as differences in the number of samples expressed the Bcl-2 and the intensity of the expression between the Iraqi and the Italian groups. Bcl-2 expression was found elevated in 26 (52%) of the 50 Iraqi breast cancer samples and 11 (36.66%) of the 30 Italian samples compared to the control samples. And in which only 1 (10%) out of 10 samples. The overexpression of Bcl-2 associated with poor prognosis for the patients. Level of Bcl-2 expression can be used as prognostic marker to monitor patient therapy. For the Iraqi cases the overexpression may be attributed to many factors like exposure to depleted uranium and bad food quality during the 90th when Iraq was under the siege of the United Nation (UN).

Abstract
Multiple studies have been reported the stimulatory effect of the combinations of nutrients factors on bone formation. One such factor is vitamin K2 which can be associated with bone protective activities. The effect of vitamin K2 alone and in combination with genistein, coumestrol and daidzein on osteoblast differentiation and mineralization were tested. Significantly, vitamin K2 increased bone mineralization in combination with genistein (10-5M), coumestrol (10-7M) and daidzein (10-5M). However, there is no additive effect of this vitamin on alkaline phosphatase (ALP) levels in osteoblasts. By contrast, vitamin K2 enhanced the stimulatory effect of type I collagen and osteocalcin expression. Vitamin K2 alone increased RUNX and OSX expression while there is no synergistic effect with tested compound; this vitamin also did not modulate nuclear factor kappa B ligand (RANKL)/ osteoprotegerin (OPG) ratio expression. These results suggested that vitamin K2 can be more effective factor in the presence of phytoestrogens on the improvement of bone formation after menopause.

Abstract
Multiple studies have been reported the stimulatory effect of the combinations of nutrients factors on bone formation. One such factor is vitamin K2 which can be associated with bone protective activities. The effect of vitamin K2 alone and in combination with genistein, coumestrol and daidzein on osteoblast differentiation and mineralization were tested. Significantly, vitamin K2 increased bone mineralization in combination with genistein (10-5M), coumestrol (10-7M) and daidzein (10-5M). However, there is no additive effect of this vitamin on alkaline phosphatase (ALP) levels in osteoblasts. By contrast, vitamin K2 enhanced the stimulatory effect of type I collagen and osteocalcin expression. Vitamin K2 alone increased RUNX and OSX expression while there is no synergistic effect with tested compound; this vitamin also did not modulate nuclear factor kappa B ligand (RANKL)/ osteoprotegerin (OPG) ratio expression. These results suggested that vitamin K2 can be more effective factor in the presence of phytoestrogens on the improvement of bone formation after menopause.

Abstract
Diarrhea is a major cause of illness and death in children worldwide; however, little information exists about the origin of childhood diarrhea in Iraq. Rotavirus, Adenovirus and Astrovirus are the major causes of sever gastroenteritis in infant and young children, pattern also observed in adult. Confirmation of viral infection by laboratory testing is necessary for reliable surveillance and can be useful in clinical settings to avoid inappropriate use of antimicrobial therapy. Methods: A total of 188 patients their age range from 1-19 (Mean=5.57 ± S.D. = 4.81) years old suffering from diarrhea were included in this study. Stool samples were collected and tested for Rotavirus, Adenovirus and Astrovirus antigens by using the rapid chromatographic test and for Rotavirus and Adenovirus Antigens, ELISA also was done. Rotavirus, Adenovirus and Astrovirus antigens were determined by rapid chromatographic immunoassay in 27 specimens (14.36%), 0 (0%) and 0 (0%) of 188 frozen stool specimens, respectively. Moreover, of these 188 specimens, Rotavirus was found in 35 specimens (18.62%) and Adenovirus in 6 specimens (3.19 %) by using ELISA technique. The present results revealed that Rotaviruses and Adenoviruses have an important role in diarrhea among children especially those less than 5 year’s old and viral pathogens should be investigated routinely in diarrhea stool specimens. This study was aimed to determine the frequency of Rotavirus, Adenovirus and Astrovirus in patients with acute gastroenteritis admitted to Al-Emamain Al-Kadhemain Medical City Hospital in Baghdad-Iraq.

Abstract
This study was designed to incorporate leishmania donovani antigens in nanoliposomes prepared by size exclusion (using Sephadex G25) and organic solvent (using Chloroform). Lipids mixture of 4Mm Phosphatidylcholine, 2.2mM Cholesterol and 0.55mM Phosphatidylethanolamine in a ratio of 7:2:1 was depended in two nanoliposome preparation methods. Physio-chemical characterizations of prepared nanoliposomes was performed by using Scanning Electron Microscope (SEM ), Fourier transform infrared spectroscopy (FTIR) and Zeta Potential assays to determine the size, morphology, chemical active group and charge . Parasite reactivation was carried out when inoculated into RPMI and incubated at 23 ̊ C for 4 days. Soluble Leishmania Antigenes (SLAs) were extracted from the promastigotes ghost membrane after fourth passages of subculturing in SNB9. The extracted SLAs were entrapped in prepared. The percentage of nanoliposomes entrapment efficiency (EE) was 62 and 50 of SLAs for chloroform and Sephadex G25 methods, respectively. Moreover, stability of SLAs entrapped nanoliposomes at 4 and 37 ̊C, as storage temperature, was examined. The stability at 4 °C showed decreasing in EE to 32 and 16 %, while stability at 37 °C revealed decreasing in EE to 16 and 8 % within 12 days of storage for nanoliposomes prepared in both methods, respectively.

Abstract
Rheumatoid arthritis (RA) is an autoimmune disease that may be triggered by urinary tract infection (UTI) especially if they are investigated in the presence of lactoferrin antibodies. The objective is to determine anti lactoferrin antibodies in RA and its association with urinary tract infection. Out of 162 autoimmunity Iraqi patients, UTI and levels of anti lactoferrin antibodies were assessed in 98 (60.49 %) with rheumatoid arthritis (RA), compared with 30 apparently healthy control and 74.48 % were observed to have UTI, the predominant bacteria identified as a cause of UTI; Proteus spp. was present as a single causative pathogen in 58.90 %, while E. coli was observed in 17.80 %. Anti lactoferrin antibodies were significantly higher in RA patients was observed in UTI+ve versus UTI-ve cases in total patients compared with healthy control that P value (≤0.05).

Abstract
The importance of Mesenchymal stem cells (MSCs) represents a favorable tool for new clinical concepts in supporting tissue engineering and cellular therapy. Bone marrow (BM) was considered important source contain mesenchymal stem cells .Another promising source of MSCs is adipose tissue (AT). MSCs derived from these sources compared regarding morphology, the success rate of isolating MSCs, expansion potential by rate of colony forming and immune phenotype. The obtained results from this study showed no obvious considerable differences concerning the morphology and immune phenotype of the MSCs derived from these sources were obvious. Differences observed concerning to the success rate of isolating MSCs, which was approximately more than 90% for BM, while it reached about 70% for AT after seven days of culturing, as well as the rate of colony forming was lower in AT cells in comparison to that obtained in BM at the same period. However, AT-MSCs could be required longest time to complete monolayer confluence, whereas BM-MSCs had the shortest proliferation period. Cells from both sources determined according to immunohistochemistry by CD105+ and CD34.¯ Conclusions revealed that MSCs can easily and successfully obtained from bone marrow and adipose tissues, and both tissues appears suitable sources of stem cells for potential use in regenerative medicine, repairing damaged tissue nevertheless the BM-MSCs more effectual in expansion and proliferation.

Abstract
This study included 50 blood samples that were collected from patients with age ranged between 35-65 years. Thirty samples were collected from patients with Type 2 Diabetes Mellitus (T2DM), while 20 blood samples were collected from healthy individuals as a control sample. The polymorphism results of TGF-β1 gene in codon 10: +869*C/T position by using amplification refractory mutation system (ARMS-PCR) showed that the T allele was suggested to have a protective effect, while C allele was associated with an increased risk of T2DM. The TT and CT were suggested to have a protective effect, while CC genotype was associated with an increased risk of T2DM. The polymorphism results of TGF-β1 gene in codon 25: +915*G/C position in samples showed that the G allele was suggested to have a protective effect, while C allele was associated with an increased risk of T2DM. The GC genotype was suggested to have a protective effect, while GG and CC genotypes were associated with an increased risk of T2DM.

Abstract
This study was aimed to isolate and identify the fungus Trichophyton violaceum from human skin specimens and evaluate the activity of antibacterial and plant essential oils. The results showed of direct microscopy by using KOH examination and morphological identification, nine samples from skin were positive in KOH test. Microscopical examination appeared that colonies of T. violaceum were coarse with abundant aerial mycelium on SDA, growth rates 6-8cm/14 days. The color changes with age from white in the middle to brownish yellow at the edges which was more intense on the reverse side. Susceptibility test to antibacterial showed that 100% of T.Violaceum T2 isolate was resistant to TE (Tetracycline), AK (Amikacin), CTX (Cefotaxime), CFM (Cefixime), CIP (Ciprofloxacin), ERY (Erythromycin) and STR (Streptomycin). Also results showed that all plant essential oils at different concentrations significantly inhibit growth of T. violaceum T2. However, the peppermint oil, Myrrh oil, Cardamom oil, Chamomile oil and Castor oil at highest concentration (25mg / ml) caused highest reduction of mycelia growth (100%) followed by Olive oil (67.3%) and Clove oil (64%) at the same concentration, compared with the control treatment

Factor V Leiden (FVL) (G1691A) and prothrombin gene (G20210A) mutations are the most common inherited forms of thrombophilia. The main objective of this study is to analyze the association between inherited thrombophilia FVL mutation and prothrombin G20210A mutation and recurrent pregnancy loss (RPL) among women suffering from complications of pregnancy. The study included 40 buccal swab samples collected from women at the reproductive age complications; abruption placenta 12.5%, dead fetal 37.5%, and recurrent spontaneous abortions (RSA) 50% in comparison with 30 women who had one or more normal pregnancies from Elwiyah Obstetric teaching hospital through the period from mid of September 2014 to the mid of March 2015. The median age of patients was 32 years (range: 19–42 years) while for the control group, it was 28 years (range: 17–41 years). Out of 40 women, 65% had one pregnancy loss and 35% for more than two previous pregnancy losses. According to the time of pregnancy losses, 22(55%) women had early pregnancy loss (EPL), and 18 (45%) women had Late Pregnancy Loss (LPL). For FVL mutation detection the restriction fragment length polymorphism (RFLP) was used. DNA fragment of interest was amplified by PCR and then subjected to digestion by MnlI specific restriction enzyme. For prothrombin G20210A mutation detection different PCR products were generated using a set of primers in A/A, G/G, A/G alleles. Out of the 70 samples tested for FVL mutation no homozygous FVL mutation was detected and Prothrombin gene mutation G20210A was totally absent among patient and control.

Abstract
Moringa oleifera is an important medicinal plant, which contains a lot of bioactive compounds. The dried plant leaves were extracted in 70% methanol by maceration using shaker incubator at 40◦C. Chemical detection of bioactive compounds in crud plant extracts was performed. The total flavonoid was isolated from the extract using reflux, The use of thin layer chromatography technique that assisted in the detection of Qurciten, Rutin and Luteoline of flavonoids isolated from the extract, the existence of qurciten and rutin confirmed by the use of high performance liquid chromatography technique. The calculation of total flavonoid of M. oleifera leaf extracts by using Spectrophotometric technique was done. The aglycon moiety was extracted by ethyl acetate, and then evaporated to dryness. The dried residue then redissolved in 50% ethanol.