Online ISSN: 2788-6867

Main Subjects : Biotechnology

Obtaining L - Glutamate Oxidase Enzyme from Streptomyces and Estimation Its Toxicity on Blood Components

Dina H. Abdul Hameed; Entesar H. Ali

Journal of Applied Sciences and Nanotechnology, In Press
DOI: 10.53293/jasn.2021.4042.1069

Streptomyces was isolated from the soil and then tested to diagnose morphologically and microscopically. Also, cultivated in the production medium of the L-glutamate oxidase enzyme, for which the wheat bran culture media was used, and then the extracellular L-glutamate oxidase enzyme was extracted using a cooling centrifuge at 8000 rpm for 20 minutes. The optimal temperature and pH for the production of the enzyme’s conditions were studied. Research results revealed that the optimum temperature for enzyme production was 30°C, and the best pH for the enzyme’s production was 7.0 later on, the enzyme was purified using common purification techniques represented by precipitation with ammonium sulfate as a preliminary purification step, followed by dialysis to remove salts, ion-exchange chromatography and gel filtration chromatography were used to finish purification process. The effect of the enzyme toxicity on the human blood components was evaluated. The enzyme had no harmful effect on the blood cells, with Red Blood Cells reaching 4 x 1012, White Blood Cells 7.5 × 109 and Platelets 293 × 109.

Evidence of Antioxidant Activity of Novel L-Glutaminase Purified from L. Gasseri BRLHM

Butheina Hasson; Likaa Mahdi; Rajwa Essa

Journal of Applied Sciences and Nanotechnology, In Press
DOI: 10.53293/jasn.2021.3969.1064

Probiotic strains have the potential to be used as bio-preservatives and functional radical scavenging treatments in the future.  Antioxidant tests, including DPPH radical scavenging, were used to evaluate the antioxidant effects of extracellular L- Glutaminase isolated from L. gasseri BRLHM. Parameters for the promoted production of the enzyme under minimal production media were optimized.    The importance of this study lies in enhancing the production of the L- Glutaminase isolated from L. gasseri BRLHM with a high activity using these L. gasseri bacterial as antioxidant activity. In ion-exchange chromatography, the specific activity was 14.7U/mg protein, with 58.8% yield and 4.6 purification folds.   In the gel filtration, the specific activity was 23.4 U/mg protein, with a yield of 54 % and 4.6 purification folds. According to the findings, L-Glutaminase isolated from L .gasseri BRLHM exhibited good antioxidant properties. As the concentration rose, there was a remarkable proportionate increase in scavenging activity. The IC50 values for control and L- Glutaminase were 36.09 1.12 and 619.8 gm/ml, respectively. The IC50 values were discovered to be 100 and 200 μg/ml, respectively. Conclusion: For the first time, the high of L- Glutaminase isolated  L. gasseri BRLHM was shown to exhibit antioxidant. This could be a promising discovery for future radical scavenging treatments and antioxidant prophylaxis with natural probiotics.

The Effect of LRP-1 Gene and Mir-205 on Cardiovascular Disease in Iraqi Patients with Familial Hypercholesterolemia

Zinah A. Ghareeb; Hiba Abdel-Hassan AlKhafaji; Mohanad K. Aneed Al-Saedi

Journal of Applied Sciences and Nanotechnology, 2021, Volume 1, Issue 3, Pages 24-31
DOI: 10.53293/jasn.2021.3691.1036

Familial hypercholesterolemia (FH) is a genetic disorder characterized by autosomal inheritance in genes related to LDL-C metabolism, with the major clinical features of hyper-LDL-cholesterolemia and premature coronary artery disease. (LRP-1) is a member of the LDLR family. It is a membrane receptor with scavenging and signaling properties. LRP-1 interacts with a wide range of extracellular ligands as well as intracellular scaffolding and signaling proteins, which makes it important in crucial clinical circumstances like cardiovascular disease, cancer, and neurological illnesses. Mir-205 uses these molecules as biomarker for cardiovascular diseases. This study aims to measure gene expression for the LPR-1 gene and its relationship to the development of cardiovascular disease in familial hypercholesterolemia and non-familial hypercholesterolemia. Also, it studies the indication whether mir-205 regulates the action of the LRP-1 gene in terms of increasing or decreasing gene expression. However, the available methods for measuring LRP1 levels are direct and quantitative using Poly Chain Reaction (RT-PCR) in real time, not at its ends. In the present study, blood was isolated from 150 individuals distributed into three groups: Group 1 included: 50 samples from a healthy group; Group 2: 50 samples from non-Familial hypercholesterolemia patients; Group 3:50 samples Familial hyperchol-esterolemia patients. The results showed that LRP1 protein expression was significantly reduced in patients with F.H compared with normal control in a small cohort from an Iraqi population. This pilot study suggests that the reduced LRP1 protein expression may be associated with cardiovascular disease progression.

Primer and Probe Designing to Detect SNP rs 4073 in Interleukin-8 Gene in Iraqi Patients with Bronchial Asthma

Media F. Ali Jan; Basima Q. Hassan Al-Saadi; Hiba M. Abdel-Hassan Al-Khafaji; Mohanad K. Aneed Al-Saedi

Journal of Applied Sciences and Nanotechnology, 2021, Volume 1, Issue 3, Pages 51-57
DOI: 10.53293/jasn.2021.3542.1034

Asthma is one of the most prevalent non-communicable illnesses, and it has a significant influence on the quality of life for many people. Asthma is now thought to be a multifaceted condition, with interactions between genetic susceptibility, host factors, and environmental exposures increasingly being blamed for its pathogenesis. In respiratory illness, interleukin-8 plays a critical function due to promoting phagocytosis and inducing angiogenesis. Also, identification and finding out the effect of these SNPs and how it can be dangerously related or contribute to a Bronchial Asthma disease. However, the available methods for detecting interleukin-8 gene polymorphism are direct and quantitative using Poly Chain Reaction (RT-PCR) in real time. Whole blood was isolated from 100 patients with asthma and 50 healthy individuals. The results of polymorphism in single nucleotides showed an essential role in the development of asthma and that the presence of these SNPs has a role in increasing the susceptibility of individuals to asthma, as the variation in the TT genotype at the site followed by the AT genotype at the same site shows high-risk damage.

Antioxidant Activities of Resveratrol Loaded Poloxamer 407: An In Vitro and In Vivo Study

Ali A. Al-Jubori; Ghassan M. Sulaiman; Amer T. Tawfeeq

Journal of Applied Sciences and Nanotechnology, 2021, Volume 1, Issue 3, Pages 1-12
DOI: 10.53293/jasn.2021.3809.1046

Resveratrol (RES) is a bioactive molecule with potent antioxidant properties, and it constitutes an essential component of herbal medicine. This study was designed to use a nano-syntheses approach to encapsulate RES in Poloxamer 407 nanoparticles. This type of nano-construction has been employed in a variety of biological applications, both in vitro and in vivo. The contribution of this nano-construct is to increase antioxidant activity over the pure RES was investigate. 1,1-diphenyl-2-picryl hydrazyl (DPPH), and hydrogen peroxide radical scavenging assays were utilized in vitro. RES-loaded poloxamer 407 showed concentration-dependent scavenging action in the range of 20-80 µ, with a maximum activity of 80.1% at 80 µ Whereas pure RES had a decrement of 61.7%. While the activity of positive control (Vit. C) was 93.2%. The magnitude of blood hemolysis examination was 3.9% at 80 µ That may indicate RES-loaded poloxamer 407 provide significant red blood cell protection. The pure RES did not prevent the appearance of abnormal cells (echinocytes), and that was cured by RES-loaded poloxamer 407. Finally, the safety of RES-loaded poloxamer 407 was assessed in vivo. Male mice were invested to detect the functions of their liver and kidney. A histopathological study was included as well. The findings showed that RES-loaded poloxamer 407 might have superior characters as a drug delivery system, nutritional supplements, and may be used in pharmaceutical products.

Correlation of MicroRNAs-122a Gene Expression with Diabetic for Iraqi Patients

Noor Younus; Maha Altaee; Zahraa Abdul-Munim Sharba

Journal of Applied Sciences and Nanotechnology, 2021, Volume 1, Issue 3, Pages 64-72
DOI: 10.53293/jasn.2021.3789.1043

This study was carried out to describe the gene expression of the micro RNA 122a gene with the development of diabetes in Iraq. The difference in gene expression between patients and healthy controls was properly considered. In this study, blood was isolated from 121 individuals divided into two groups as follows: 80 samples of diabetic patients and 41 samples from a healthy control. miRNA was isolated and transformed into cDNA, and the expression of mi122a was measured by qRT-PCR. The researchers looked at the relationship between age and gender and the occurrence of diabetes, as well as how they compared to controls. When comparing the mean gene expression level (Ct) of patient groups to the corresponding Ct means in the control group, the results revealed a discrepancy. Also the gene expression folding (2-∆∆Ct) of the micro RNA 122a gene reflect differences in the expression, the level of micro RNA 122a was (20.504) in patients with diabetes compared to control groups with significant differences. On the other hand, gender and family history showed a significant difference between patients and health monitors. For age and type of diabetes, they showed a significant difference between patients and health monitors. Our results indicate that diabetes can affect all ages in both males and females. This study aims to correlate the expression of miRNA 122a with the occurrence of diabetes in the Iraqi population.

Evaluation The Antibacterial Activity of Biosynthesis Silver Nanoparticles by Lactobacillus Gasseri Bacteria

Raghad Jabbar; Nehia N. Hussein

Journal of Applied Sciences and Nanotechnology, 2021, Volume 1, Issue 3, Pages 86-95
DOI: 10.53293/jasn.2021.3898.1057

Biosynthesis of AgNPs is a new approach in the field of nanotechnology with optimistic implementation in medicine, food control, and pharmacology. In this study, the silver nanoparticles were produced by Lactobacillus gasseri filtrate. The production of AgNPs was confirmed by the color change from yellow to brown. Using UV visible spectrophotometer at 424 nm wavelength, and X-ray diffraction, it was found that the size of the synthesized particles was 58.5 nm after applying Scherrer’s equation. The inhibitory activity of silver nitrate on the growth of some pathogenic isolates was studied Staphylococcus haemolyticus Gram positive, and Klebsiella pneumoniae Gram negative. The highest inhibitory diameter was 14.6 mm at 100% concentration (stock) against Staphylococcus haemolyticus bacteria was followed by that of Klebsiella pneumoniae bacteria with an average inhibition zone diameter reached 13.6 mm at 100% concentration. The highest effect was of AgNPs on the growth of Staphylococcus haemolyticus bacteria, as it found the average diameter of the inhibition zone reached to 29.3 mm, followed by Klebsiella pneumoniae with the average diameter of the inhibiting zone it was 22.6 mm at 100% concentration (stock). This study showed AgNPs have more antibacterial activity against Gram positive bacteria than Gram negative bacteria. The importance of this study lies in testing the effectiveness of by Lactobacillus gasseri bacteria in the biosynthesis of silver nanoparticles and studying their antibacterial activity on pathogenic bacteria.

Characterization of Treated Multi-Walled Carbon Nanotubes and Antibacterial Properties

Noor Ali; Ali Taha; Duha S. Ahmed

Journal of Applied Sciences and Nanotechnology, 2021, Volume 1, Issue 2, Pages 1-9
DOI: 10.53293/jasn.2021.11636

In the present study, the synthesized Multi-Walled Carbon Nanotubes (MWCNTs) were chemically treated with a mixture of acids to produce functionalized MWNTs. The functionalized MWCNTs were characterized by X-Ray Diffraction Analysis (XRD), Zeta potential and Field-Emission Scanning Electron Microscopy (FE-SEM). The X-ray diffraction reveals the MWCNTs average crystal size of the R-MWCNTs and F-MWCNTs were about (3.27 and 3.19) nm, respectively. FESEM images show the formation of R-MWCNTs that appears as cylindrical tubes and aggregated tubes with each other, while the F-MWCNTs appear as less aggregated and tangled clusters than R-MWCNTs. Zeta potential measurements showed that the measurement of the R-MWCNT shows a low negative value -9 mV and F-MWCNT, it was found that the zeta potential value is up to -29 mV. The antibacterial activity was studied against E. coli and P. aeruginosa bacteria, and indicated the highest growth inhibition zones (IZ) of F-MWCNTs as compared with R- MWCNTs against E. coli and Pseudomonas aeruginosa, respectively.

Enhanced Production of Fibrinolytic Enzyme from Pseudomonas aeruginosa by Optimization Media Components

Bilal Jasim; Entesar Ali

Journal of Applied Sciences and Nanotechnology, 2021, Volume 1, Issue 2, Pages 58-65
DOI: 10.53293/jasn.2021.11637

Fibrinolytic enzyme is factor that lysis fibrin clots. This fibrinolytic factor has prospective use to treat cardiovascular diseases, such as stroke and heart attack. Cardiovascular diseases attracted worldwide attention for their elevation morbidity and mortality. Expensive cost and fatally undesired side effects associated with the available fibrinolytic agents to treat these diseases stimulated the researchers to investigate potentially better agents for curative applications. In the current investigation, fibrinolytic enzyme production from Pseudomonas aeruginosa isolated from injuries of wounds and burns patients. Parameters for the promoted production of the enzyme under minimal production media were optimized. It comprised carbon source (glucose), Nitrogen source (Yeast extract), Fibrinogen concentration (0.5 %), inoculum size (1 %), temperature (37°C), and PH (7). Enhanced fibrinolytic enzyme activity (136.2 IU/ml) was obtained after optimization Medium Components compared with that obtained with the minimal medium (60.2 IU/ml) which is 2.2 times higher than the same under non optimized production conditions. Media optimization researches for enhancement of fibrinolytic enzyme production from Pseudomonas aeruginosa in Iraq has not been performed so far. This may be the first study to optimization media for the production of fibrinolytic enzyme from Pseudomonas aeruginosa. The importance of this study lies in the enhancing the production of the fibrinolytic enzyme with high activity using these bacteria.

Evaluation of the Biosynthesized Silver Nanoparticles’’ Effects on Biofilm Formation

Nehia Hussein; Mina M. Khadum

Journal of Applied Sciences and Nanotechnology, 2021, Volume 1, Issue 1, Pages 23-31
DOI: 10.53293/jasn.2021.11019

In this study 50 isolates were obtained from the Baghdad teaching city medicine laboratories, from wounds and burns. Isolates were identified exercise VITEK 2 system (Biomerieux). Streptococcus pyogenes isolate was used to create the biosynthesize of silver nanoparticles’’ against some pathogenic microbes such as Pseudomonas aeruginosa, Staphylococcus aureus, Escherichia coli, and Candida albicans. Evaluation of the effect of the created biosynthesis silver nanoparticles’’ (AgNPs) by Streptococcus pyogenes on the biofilm formation by various human pathogenic bacteria.  Biosynthesis of AgNPs was characterized by ultraviolet-visible absorption spectroscopy, the observation of color change of the experimental samples in the presence of 1 mM AgNO3 at 410 nm. A color change from pale yellow to slightly brown occurred for bacterial supernatant within 24 hours of incubation in the presence of light Scanning electron microscope (SEM), the biosynthesis silver nanoparticles’’ are predominately circular fit as a fiddle having a smooth surface and very much scattered with close minimal game plan, X-ray diffraction (XRD). The The normal molecule size was determined by Debye-Scherer equation and its evaluation was roughly 6.43nm. The normal molecule size was determined by Debye-Scherer equation and its evaluation was roughly 6.43nm. The importance of this work lies in the possibility of synthesizing the silver nanoparticles’’ using these bacteria, which are considered as types of fastidious bacteria. As far as the researcher’s knowledge is concerned, this is study is the first of its kind in Iraq.