Journal of Applied Sciences and Nanotechnology

Covid-19 is a global challenge that drives health care to its limits. Biological parameters associated with increased risk of severe or fatal disease courses were identified, including conditions connected with a poor state of health, organ damage, and coagulation dysfunction. In a recent study, risk factors like age, sex, blood group (ABO), smoking, and several biomarkers like white blood cells, lymphocytes, C-reactive protein (CRP), ferritin, D-dimer, and potassium level were considered. One hundred nasopharyngeal swabs and blood samples were collected from Iraqi patients and classified according to the severity of the disease into five groups: Asymptomatic, Mild, Moderate, Severe, and Deceased, according to the recommendations of the World Health Organization (WHO). The current study was based on the risk factors and their effect on disease severity (age, sex, blood type, and smoking). As a result, there was a substantial positive linear link (R2=0.91) between getting older and increasing disease severity. Study results were showed a little statistically significant difference in the number of males and females infected with SARS-CoV-2. Interestingly, males were shown to be more susceptible to infection than females. Also, because type (O) blood is more extensively distributed in the Middle East population than the other blood types, those with type (O) blood are more susceptible to infection. Because smokers and non-smokers were distributed at random across different disease severity levels, there was no statistically significant link between smoking status and COVID-19 severity. All parameters (plasma protein and liver enzymes) showed statistically significant differences, especially between the deceased groups except K level, which did not differ significantly in all groups. This study aimed to investigate some biological markers during infection to see if there was a link between these measurements and risk factors, as well as the severity and progression of the disease.

In the current study, bioinformatics approach has been utilized to discover the sequences and structures analysis of IL-17A of breast cancer and compares with normal sequence from NCBI. The current study was aimed to discuss the possibility of using IL-17A as a marker for patients with breast cancer. Also, the effect of mutation on the physicochemical properties and structure of IL-17A. Sixty blood samples were examined from patients with breast cancer (aged between 20 and 75 years old). In patients with breast cancer, there were missense mutations and deletion mutations detected by BLAST. Furthermore, the current study determined the physicochemical properties of IL-17A, such as their hydrophilic nature; alpha–helical structure, and 3D structure. The results of this study show that IL-17A consider as a marker for the patient with breast cancer. Also, the mutations on IL-17A gene affected the structure and physicochemical properties of the Interleukin-17A protein complex.

Pseudomonas aeruginosa is an opportunistic pathogen that can cause different infections such as nosocomial bacterial infection, Urinary tract infection, Meningitis, Eye infection, Otitis Media, Respiratory infections, and Pneumonia, especially in people with Cystic fibrosis, Bone and joints infection, gastrointestinal infection, Skin infection and soft tissue inflammation.  Resistance of P. aeruginosa to antibiotics and disinfectants is a major problem all over the world due to the random and unrestricted use of antibiotics. This resistance is also likely to occur due to several reasons, including the production of beta-lactamase enzymes and the lack of permeability of their cell wall, possibility of it containing resistance genes and the occurrence of genetic mutations leading to the development of resistance to antiseptics and causing failure in treating infections that caused by P. aeruginosa. This study aims to detect and sequence analysis of qacE gene in P. aeruginosa that responds to the resistance of Quaternary ammonium compounds and detect the mutations in this sequence and detection protein synthesis in this gene.  A hundred swabs were collected from patients with burns and wounds infection from many Hospital in Baghdad including Al- Kindi Teaching Hospital, Al-Yarmouk Teaching Hospital, Baghdad Teaching Hospital, National Centre for Educational Laboratories, Al-Shahid Ghazi Hariri Specialist Specialized Burns Centre (Medical City), and Al-Zaafarania General Hospital during the period October 2017 for the end of December 2017.The current study detected 69 isolates of P. aeruginosa from wound 68.6% (24/35) and burn samples 69.2% (45/65). The study was investigated QacE gene in 97.1% that responses of P. aeruginosa resistance to Quaternary ammonium compounds. The sequencing analysis for QacE genes were determined and the results appeared multiple mutations including Missense mutations, insertion mutations and several silent mutations, some mutations effected in the translation of protein while others not affected. This study also included the analysis of phylogenetic tree of QacE gene and the results showed that the tree consisted of only one species which is P. aeruginosa, and the isolates in this study did not deviate of this bacteria even if their mutations were more than 22.

Klebsiella pneumoniae is a noteworthy human pathogen. As a virulence factor, these bacteria may create a thick coating of extracellular biofilm. This aids the organism's adhesion to biotic as well as abiotic surfaces, preventing antimicrobial agents from doing their job. Infections caused by bacterial biofilms have become more difficult to treat as a result. Therefore, the present study has been designed to investigate the effects of essential oils, individually or in combination, on the biofilms of Klebsiella pneumoniae isolates. In this research, the quantification of biofilm composition for 50 isolates from urine samples indicates the following statistics: [n = 24 (48%)] isolates form a strong biofilm, [n = 12 (24%)] a moderate biofilm, [n = 10 (20%)] a weak biofilm, and [n = 4 (8%] a non-biofilm. The Minimum Inhibitor Concentration (MIC) and MBC values for essential oils were determined. The results showed that the MIC for tea tree oil was 0.25% and MBC 0.5%. While the MIC for cinnamon oil was 0.125%, and MBC was 0.25%. Afterward, the anti-biofilm effectiveness of essential oils was evaluated. The results showed that both oils had good efficacy against strong biofilm for Klebsiella pneumoniae isolates. But in a comparison between them, cinnamon oil showed better results. Due to the efficacy of these two oils, the combined impact of the two oils was discovered in this study. And the results revealed that there was an antagonistic effect. These findings recommend additional essential oils be tested to see how they affect biofilms of Klebsiella pneumoniae or other bacteria.

Interferon gamma (IFN-γ) is a cytokine involved in the induction and modulation of various immune responses. INF-γ has recently been associated with breast cancer and the stage of the disease. Also, it can be used as a cancer marker in determining disease activity. The ELISA test is a technique for identifying antigens in biological materials. The ELISA test is used for the detection of antibodies against target antigen through highly antibody-antigen interactions. In this study, the blood samples were taken from 88 Iraqis, 30 healthy controls, and 58 breast cancer patients (ranging from 35 to 52 years of age). During a visit to the AL Amal National Hospital for Cancer Treatment in Baghdad, Iraq, these patient samples were taken October 2020 - January 2021. Depending on the color changes to yellow, the results of this study were determined under wavelength of 450 nm to reveal the optical density (OD) via spectrophotometrically. The OD value is proportional to the amount of human IFN-γ in the sample of the healthy control and the cancer patients. The blood concentration of IFN-γ was substantially greater in women with breast cancer than in the control group (43.15 ± 4.68 pg/ml vs. 18.8 ± 3.89 pg/ml), respectively. Our results showed an elevation of Interferon-gamma (IFN-γ) in most of the serum samples from women with breast cancer. The conclusions of this study are the main findings, INF-y protein was employed as a marker and assistance in diagnosing breast cancer based on the results of the calculation of protein concentration from the ELISA test.

In this paper, the study aimed to analyze the sequence and structure of Transcription Factor 7 Like 2 gene (TCF7L2) in Iraqi Diabetic Mellitus Type II (T2DM) Patients and was compared with standard sequence form National Center of Bioinformatics Tools (NCBI) using BLAST. Ten blood samples of (T2DM) Iraqi patients, was collected from Al-Mustansiriya University National Diabetes Centre in Baghdad Province/Iraq (17-65 year). DNA was extracted from whole blood of patients’ samples using the Quick-DNA™ Blood MiniPrep kit, then it was sent to Korea at Macrogen Corporartion Company, where they used automated DNA sequencing for sequence analysis. The results of the sequence analysis of (T2DM) patients’ samples, we found: six missense mutations, one deletion mutation, and three silent mutations detected using BLAST in (NCBI). All mutations appeared at the same sites of the gene which controls the rate of genetic information transcription that indicates to have a relationship with (T2DM). These mutations were recorded on the (NCBI). The physicochemical properties of (TCF7L2) determined in the present study included; alpha-helical structure and 3-Dimension structure appeared contrast when compared with the gene template. Briefly, mutations effected (TCF7L2) which influences the structure, physicochemical properties of the protein, and the secretion of insulin hormone which maintains glucose level in blood.

This study was carried out for the mycosynthesis of silver nanoparticles by Candida albicans supernatant.  All the isolates used in this study were taken from the patients who existed at Al-Elweya children's teaching hospital in Baghdad, Iraq. Mycosynthesized silver nanoparticles were characterized by color visualization, ultraviolet-visible (UV) spectroscopy, and Fourier transform infrared spectroscopy (FTIR). The UV-Vis spectroscopy examination has shown the highest absorbance (λmax) at the wavelength of 429 nanometers, which indicated the creation of silver nanoparticles. Furthermore, the results of the antibacterial potential of AgNO₃ and AgNPs against Klebsiella pneumoniae and Staphylococcus haemolyticus bacteria showed the highest effect of AgNO3 against Staphylococcus haemolyticus when the diameter of the inhibition zone reached (14.00 mm). In contrast, the lowest effect of the AgNO₃ was with the diameter of the inhibition zone that reached (11.66 mm). The highest effect of the AgNO₃ against Klebsiella pneumoniae by the diameter of the inhibition zone was reached (12.66mm), while the lowest effect was (9.00mm). The highest effect of the AgNPs against Klebsiella pneumoniae by the diameter of the inhibition zone reached (16.00mm), while the lowest effect was (13.00mm). The highest effect of the AgNPs against Staphylococcus haemolyticus by the diameter of the inhibition zone was reached (17.33mm). Where the lowest effect by the diameter of the inhibition zone was reached (14.00mm). Interestingly, this revealed that Staphylococcus haemolyticus was more susceptible to silver nitrate (AgNO₃) and silver nanoparticles (AgNPs) than Klebsiella pneumoniae.

In patients with diabetes, chronic inflammation is characterised by the increase in C-reactive protein (CRP) and tumour necrosis alpha (TNF-α). The consumption of melatonin and flaxseed oil may improve insulin sensitivity and the effectiveness of diabetic medications. The current study investigated the efficacy of melatonin and flaxseed oil in patients with Type 2 diabetes mellitus (T2DM). It involved 43 patients with diabetes who were divided into 3 groups. The first group received a placebo (starch 50mg, n=13), the second group received melatonin (10mg per day, n=14) and the third group received flaxseed oil (1000mg per day, n=16), in addition to prescribed hypoglycaemic medication and a 12-week controlled diet. Fasting blood sugar (FBS), glycated haemoglobin (HbA1c), oxidative stress and inflammatory parameters were measured in each group at 0, 6, and 12 weeks. Melatonin and flaxseed oil administrations resulted in a highly significant increase in glutathione (GSH) levels, a significant decrease in malondialdehyde (MDA), a significant increase in superoxide dismutase (SOD) in the melatonin group and a significant increase in SOD in the flaxseed oil group. The melatonin group and the flaxseed oil group also showed a highly significant decrease in oxidised low-density lipoprotein (ox-LDL).TNF-α was significantly reduced after the respective consumption of melatonin and flaxseed oil. Furthermore, flaxseed oil consumption resulted in a significant decrease in CRP; however, there was no significant difference in CRP due to melatonin consumption.

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.

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.

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 µg.ml^-1, with a maximum activity of 80.1% at 80 µg.ml^-1. 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 µg.ml^-1. 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.

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.

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.

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.

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.

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.

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.

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.