First pages:

التاثير التثبيطي لبعض المستخلصات على فعالية انزيم Angiotensin converting enzyme وبعض المؤشرات الحيوية المساهمة في ارتفاع ضغط الدم == Inhibitory Effect of Some Plant Extracts On Angiotensin Converting Enzyme Activity And Some Biochemical Marker That Associated With Hypertension

Author name: رؤى اياد يوسف

Supervisor name: غازي منعم عزيز | حسن فياض

General topic: Biology

Specific topic: Biotechnologies

Degree: Master

University: University of Baghdad - College Of Science - Department Of Biotechnology

Language: English

University location: Baghdad

First pages:

Abstract: The present study was sought to investigate the inhibitory effect of four crude aqueous plant extracts : Zingiber officinale Roscoe (ginger), Camellia sinensis (Green tea), Olea europaea (Olive) and Hibiscus sabdariffa (Rosella) on key enzyme linked to hypertension, Angiontensin - I Converting enzyme (ACE), and on the oxidant/antioxidants status, lipid profile in vitro and in vivo studies. Study of some biochemical biomarker demonstrated that antioxidant enzyme, oxidant enzyme, liped profile and ACE level for 75 hypertension patients. Antioxidant enzymes superoxide dismutase (SOD) and glutathione peroxidase (GPx) were shown to have cross relationship with ACE level in hypertension groups, while lipid profile have positive relationship with ACE level in hypertension groups. ACE activity for the four groups G1, G2, G3, G4 were 19.61±3.97, 21.3 ± 1.95, 28.06 ± 5.34 and 32.74±8.19 ng/ml respectively. From these results we concluded that ACE was associated with hypertension and its modulated by drug or herbal extracts. Angiotensin - I Converting enzyme was extracted from sheep lung with specific activity 0.08 U/mg, then the crude ACE extract was concentrated with sucrose by dialysis with specific activity 0.1 U/mg, purification fold was 1.25.The enzyme was purified partly by ion - exchange chromatography using DEAE - cellulose with specific activity 0.5U/mg, yield 30% and purification fold 6.25. ACE activity was determined using N - [3 - (2 - furyl) acryloyl]L - phenylalanyl glycyl glycine (FA - PGG) as a substrate. Results for in vitro ACE inhibitory activity using plant extracts (ginger, green tea, roselle and olive) showed that the all four crude aqueous plant extracts had inhibitory activity in different values when used in the same concentrations about (1 mg/ml), and ginger extract possessed higher inhibitory activity than other three extracts. The ACE inhibitory potency of the ginger extract was found to be significant (P<0.001) when compared with the standard anti - ACE inhibitor drug (Captopril) at the same concentration. The inhibitory activity of ginger extract with different concentration (25, 50, 100 mg/kg) in L - N - ? - nitro - L - arginine methyl ester (L - NAME) induced hypertensive mice was evaluated. Acute oral administration with L - NAME 50mg/kg.b.w causes a rise in blood pressure in normal mice. Administration of aqueous ginger extract (25, 50 and 100 mg/kg) for 4 weeks in L - NAME treated mice significantly (P<0.05) reduced the mean arterial blood pressure compared with L - NAME animals without treatment, with decreasing the serum levels of ACE; while the activity of superoxide dismutase (SOD) and glutathione peroxidase(GPx) showed a significant elevation in ginger treated L - NAME induced hypertensive mice. The results suggest that ginger extract could prevent the development of high blood pressure induced by L - NAME probably can be attributed to prevent or reduce the oxidation process and the inhibition of physiological processes of a substance L - NAME and so as it contains ginger compounds of polyphenols, which inhibits the activity of the ACE and prevent oxidation of fats and repair System Antioxident. Our study concluded that ginger might act as a natural alternative to better and safer in the prevention of negative impacts and risk factors such as high blood pressure and lipids.

تاثير انزيم SNase المنقى من العنقودي في الغشاء الحياتي للايشيركية القولونية والكليبسيلا الرئوية == Impact of Snase Purified From Staphylococcus Aureus On Biofilm of Klebsiella Pneumoniae And Escherichia Coli

Author name: هند تحسين ابراهيم

Supervisor name: حارث جبار فهد المذخوري

General topic: Biology

Specific topic: Microbiology

Degree: Master

University: University of Baghdad

Language: English

University location: Baghdad

First pages:

Abstract: SDS - PAGE showed that a single sharp band with an approximately 16.8 KDa molecular weight has been seen. A matter indicates that the enzyme is consistently pure. PCR technique was applied to approve the existence of nuc gene in S.aureus Nevertheless, only those which depicted positive results on DNase agar harboured nuc gene, as it is specified by single band appearance of nuc at assumed molecular weight (ca. 255 pb) of marker that being used. The current study findings highlighted the participation of SNase purified from S. aureus in significant (P< 0.05) preventing biofilm formation by E. coli and K. pneumoniae compared with untreated controls. Evidently, the inhibitory effect of SNase on biofilm formation is undoubtedly perceived, indicating the degradation of the structural major component of biofilm formation (i.e. extracellular DNA). Results revealed that SNase was able to significantly (P< 0.05) reduce the number of the uropathogens; E. coli and K. pneumoniae attached to the uroepithelial cells. 1 Chapter one : Introduction and literature Review 1. Introduction and Literature Review 1.1. Introduction Staphylococcus aureus is a persistent human pathogen that is responsible for a range of diseases that vary widely in clinical presentation and severity. The pathogenesis of S. aureus infection is a complex process involving a diverse array of secreted and surface - associated virulence determinants that are coordinately expressed at different stages of infection (Loughman et al., 2009). Escherichia coli is a genetically diverse species that causes a variety of infections which fulfill many or all of the proposed criteria for biofilmassociated infections (Kaper et al., 2004). Klebsiella pneumoniae is an opportunistic pathogen responsible for a wide range of nosocomial infections. One important factor associated with virulence in K. pneumoniae is its capacity to adhere to surfaces and form biofilms (Cruz et al., 2012). Bacteria often exist as sessile communities called biofilms which are exquisite structures caused by a genetically programmed developmental process. It is estimated that biofilms are involved in 65% of human bacterial infections, since cells in biofilms are 1000 times more resistant than cells in the planktonic state, making medical treatments fail (Shal? et al., 2011). Extracellular DNA (eDNA) plays a significant role in biofilm formation, as revealed by studies in several bacteria including E. coli (Nakao, 2012) and K. pneumoniae (Whitchurch et al., 2002); however, there is no definite proof on a cause - and - effect relationship between DNA release and biofilm formation (Beenken et al., 2012) or becomes a key component of the macromolecular scaffold in many different biofilms (Jakubovics et al., 2013). In rare cases eDNA has been shown to inhibit bacterial settlement. Therefore, it is possible that nuclease mediated eDNA degradation would therefore promote adhesion. Consequently, it is not clear whether microbial nucleases contribute to the gross biofilm structure in clinically relevant situations (Sheilds et al., 2013). 2 Chapter one : Introduction and literature Review All previous studies used both commercial bovine and recombinant human DNaseI in the disruption of medically important biofilms; whilst, extracellular nuclease of S. aureus (SNase) was used against biofilm of S. aureus themselves (Benenken et al., 2012b ; Kiedrowski et al., 2014). Thus, to date, the role of S. aureus exonucleases in biofilm of other bacteria remains unclear. However, in the present study SNase is used against biofilm of other bacterial species (viz. K. pneumoniae and E. coli). To address this, the following steps were undertaken : 1 - Investigating the negative impact of SNase extracted from S. aureus on K. pneumoniae and E.coli biofilm formation. 2 - Studying the preventive activity of SNase on the adherence of K. pneumoniae and E. coli on uroepithelial cell.

Author name: ايمان نعمان اسماعيل

Supervisor name: نورية عبد الحسين علي | مجيد ارشيد سباح

General topic: Biology

Specific topic: Biotechnologies

Degree: Master

University: University of Baghdad

Language: Arabic

University location: Baghdad

First pages:

Abstract: نفذت الدراسة الحالية في مركز بحوث التقنيات الاحيائية/ جامعة النهرين للمدة 2011 - 2012 لغرض الكشف عن جين الملوحة TaGSK1 ودراسة التعبير الجيني له في صنفين من الحنطة فرات ودجلة والتركيب الوراثي 2H المنتخبة لصفة تحمل الملوحة مقارنة مع الصنف تموز2 الحساس للملو | This study was conducted in biotechnology research center, Al - Nahrain university in 2011 - 2012 to detect the gene responsible for salt tolerant (TaGSKI) and study its expression in two wheat (Tigris and Euphrates) cultivars and the genotype (2H) were selected for salt tolerance through improvement and breeding programs as compared with to the sensitive local wheat cultivar (Tamooze 2). Seeds of the these cultivars were sown in flasks that contained Ms media in three salt levels (0, 15, 25) ds/m with three replication for each. Five seeds from each cultivars were sown in each replicate. After 10 - 15 days from the sowing, the percentage of germination was estimated, and samples of leaves were collected for RNA extraction and then changed to the cDNA. B - actin gene that consider endogenous control and salt tolerant gene TaGSK1 were amplificated by using PCR technique to discover the gene and by QPCR to estimate gene expression by determine the CT (Cycle Threshold) value for B - actin and TaGSK1 genes in wheat plants of the studying cultivars. Number of DNA dilutions of 189bp molecular weight that extracted from agarose gel were used to determing CT value for each dilution. Standard curves were drawn to find out value of PCR Efficiency which was used for gene expression for the salt tolerant gene TaGSK1.The results of germination percentage (%) showed that there were high differences between the two salt tolerant cultivars (Tigris and Euphrates) and 2H genotype and local cultivar (Tamooze 2). Euphrates cultivar gave 100% germination percentage, and the Tigris cultivar and 2H genotype gave 89, 86% germination percentage respectively at 25ds/m. whilst the local cultivar gave zero germination percentage at the same level of salinity. At 15ds/m level, also the Euphrates cultivar gave 100% germination percentage while the Tigris cultivar and 2H genotype gave 96, 94% germination percentage respectively as compared to Tamooze cultivar that gave 13% germination percentage. All the cultivars have 100% germination percentage at 0.0ds/m level. The conclusion of this result is the two cultivars (Tigris and Euphrates) and 2H genotype more salt tolerance than local cultivar at this growth stage which more salt sensitive than others growth stages. The results of PCR reaction were also indicated that Tigris and Euphrates cultivars and 2H genotype have salt tolerant gene TaGSK1, while this gene did not exist in the local cultivar. There were two bands of TaGSK1 gene that have 189bp and 404bp molecular weight in the Tigris and Euphrates cultivar and the genotype 2H, while the local cultivars have only one band that have 404bp molecular weight. These results were indicated that 189bp molecular weight of this gene is responsible for salt tolerance character in these cultivars.The results of QPCR reaction also were revealed that there is difference between the cultivars in their gene expression. Tigris and Euphrates cultivars and 2H genotype gave the highest gene expression at 15ds/m and increased at 25ds/m as compared to 0.0ds/m. At the second level 15ds/m the gene expression of the two cultivars and genotype was 0.8682, 0.8190 and 0.8688 respectively, and at the third level 25ds/m was 1.656, 1.3176 and 1.2665 respectively, while at the first level 0.0ds/m was less than the other 15, 25 ds/m as compared to local cultivar that the gene salt tolerant (TaGSK1) have no gene expression at the same salt levels. This result indicated that the local cultivar does not have salt tolerant gene TaGSK1. From these results we can revealed that TaGSK1 gene was found in the Tigris and Euphrates cultivars and 2H genotype, and this gene can be considered from high salt tolerant gene, because it gave the highest gene expression at the highest salt level 25ds/m. Therefore this gene help the plant to tolerate salt stress and grow very well. The results also showed that the two salt tolerant cultivars (Tigris and Euphrates) and the salt tolerant genotype (2H) proximately have the same salt tolerance degree, so they have proximately the same gene expression as compared to salt sensitive local cultivar which have no salt tolerant gene TaGSK1. This gene is good indicator for salt tolerance at high salinity levels 15, 25 ds/m in these cultivars and genotype.