Faculty Of Pharmacy Graduation Project 2023 -2024
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Item Analysis of laboratory biochemical results using AI aided model for Hepatocellular Carcinoma Diagnosis and Prediction(October university for modern sciences and arts, 2023) Abdelwahab, Aya Mohamed; Ragab, Mariam Khaled; Hafez, Mina Hany; Fawaz, Salma AshrafThe aim of this study is to correlate between both Long noncoding RNAs (LncRNAs) and hepatocellular carcinoma (HCC) with an attempt of integrating them in the diagnosis protocol for HCC with the help of an artificial intelligence models. samples were collected and prepared by third partner SHEFAA ALORMAN hospital. Then RNA was extracted using specific extraction kits. Quantitative polymerase chain reaction (qPCR) was used later to determine specific RNA concentration in previously prepared samples. Different models were created using different trial of several data sets. The data sets were integrated into several algorithms, the non-AI traditional results showed low accuracy but by integrating the artificial intelligence in the diagnosis it enhanced the accuracy. The data was presented as mean ± SD where the results of LINC00853 were 29.92 ± 0.006711 with sensitivity and specificity of 97.14% and 95.71% respectively. The results of HULC were 20.49 ± 11.29 with sensitivity 95.71% and specificity 94.29%. Firstly, a model was built by using traditional data (ALT, AST, Total bilirubin and Serum albumin) and showed a higher accuracy than traditional results. Secondly by the evaluation of the value of addition of AFP to the previous parameters and it was d found that the accuracy increased when compared to the model that was trained with the traditional data only. Finally, in order to increase the accuracy of the data integration of novel LncRNAs (Highly up regulated in a liver cancer (HULC)-long intergenic non protein coding 853(LINC00853)) was done where it showed promising result. To conclude the novel LncRNAs biomarkers confirmed their potentiality as a diagnostic tool for HCC diagnosis and their integration in AI model increased the sensitivity and specificity for the diagnosis when compared to traditional data alone.Item Investigating pathogenesis and risk factors of autism and its impact on clinical outcomes in Autistic Egyptian Children(MSA University Copy Right 2023, 2023) Said Shalaby, Aya; Ragaey Borik, Omar; Mohamed Zolfkar, Sarah; Magdy Sakr, TasnimIn recent years, there has been growing interest in the potential role of the gut microbiome in the development of autism. The gut microbiome refers to the trillions of microorganisms that live in the digestive tract, including bacteria, viruses, and fungi. These microorganisms play a crucial role in various physiological processes, including digestion, immune function, and brain development, beside that studies have shown that individuals with autism tend to have altered gut microbiomes compared to typically developing individuals. Specifically, they have been found to have lower levels of beneficial bacteria and higher levels of potentially harmful bacteria in their gut microbiomes. This study is a case control study design depending on pilot study of 20 participants divided into 10 control and 10 autistic patients. Our aim is to elucidate the changes in gut microbiome together with clinical variables in Egyptian autistic children and their possible correlation with the incidence and the severity of autism. As a primary objective is looking for clinical and microbiome risk factors correlated with the incidence and severity of autism as (prenatal factors, natal factors and post natal factors), meanwhile the Secondary Objectives is to detect other clinical risk factors.it gave us in autistic children the mean (±SD) age was 4.7 (±2.0) years, gestational age mean (±SD) was 38.2± (0.63) weeks, birth weight mean (±SD) was 2.99 (±0.46), and the mean (±SD) of CARS test was 36.95±4.65. Finally, there is correlation between some clinical risk factors as (GIT disturbance, Eating Carbohydrates was significant with P-value<0.001 between the two studied groups while eating protein between the levels of severity with p-value<0.05) to autism spectrum disorder (ASD)Item Novel Therapy for Inflammatory Bowel Disease(October university for modern sciences and arts, 2023) Ahmed, Omar Ashraf; Ahmed, Nourhan Abdelhameed; Ali, Nada Ahmed; Fawzy, Fawzy AshrafIrritable bowel disease (IBD) is episodes of inflammation in the gastrointestinal tract, these episodes is caused by abnormal immune response to gut microflora. IBD is divided into two types: ulcerative colitis and crohn’s disease. The causes of IBD are increased permeability of intestinal barrier and imbalance in gut microbiota. Treatment known for IBD are anti-inflammatory drugs like corticosteroids and sulfasalazine, Immunosuppressive drugs like Imuran. Probiotics are foods or supplements that contain live microorganisms intended to maintain or improve normal microflora in the body. Therefore, the aim of study is investigation the efficacy of a novel strain of probiotics to be used as a potential treatment for IBD. While, the objectives are induction of inflammation on rats and mice, evaluation of novel therapeutic agent, and assessment of its anti-inflammatory activity histopathologically and by measuring inflammatory biomarkers. We worked on 3 colitis models: first model is BALB/c mice and colitis are induced by shigella toxin, second and third models are Wister albino rats and colitis is induced in one of them by acetic acid and in the other by dextran sulfate. We made protection with our novel therapy (lactobacillus) and commercial drug (enterogermina) or sulfasalazine. After the induction step, all mice and rats were scarified, and blood samples were collected to measure inflammatory biomarkers by ELISA and Western blotting, also intestine is isolated to be examined histopathologically. Histopathological examinations of acetic acid and Shigella models showed normal mucosa without alteration in the groups protected by lactobacillus, sulfasalazine and in control group, while the group protected by enterogermina showed mild edema. Elisa results proved that lactobacillus has anti-inflammatory activity as it showed inhibition in levels of inflammatory biomarkers after induction by acetic acid and shigella. In conclusion, lactobacillus showed more anti-inflammatory activity than enterogermina, so it is a promising drug in IBD treatment.Item A Promising Ocular Delivery Approach of a Novel Nano-based Multifunctional System(October university for modern sciences and arts, 2023) Adel, Mariam; Refaat, Haidy; Safwat, Seifeldin; Mahmoud, MariamAim: Delivery of ophthalmic drugs to the interior parts of the eye remains a huge challenge because of the well-known static and dynamic ocular barriers. Nanoceria (NC), exhibits unique auto-regenerative antioxidant capacity, leading to its wide applications in the management of various oxidative stress-related diseases, especially ocular diseases. Besides, NC could be adopted as a nanocarrier to deliver specific drug to ocular tissues. Herein, the current study presents a novel multifunctional Metformin-loaded cerium oxide nanoparticles with potential synergistic ophthalmic activity for topical ocular delivery in a challenging drug repurposing approach for intraocular targeted and sustained delivery of metformin (MET), that possesses considerable anti-angiogenic and anti-inflammatory effects on retinal vasculature. Methodology: Glycol chitosan-coated negatively charged NC particles were developed and loaded into an in-situ gel/film-forming system to overcome the inconvenience and eye protective mechanisms that limit the ocular availability after the topical application of traditional ocular delivery systems. The developed particles were evaluated for entrapment efficiency (EE%), particle size (PS), zeta potential (ZP), Fourier-transform infrared spectroscopy, morphology, in vitro release, and in vivo ocular hypertension model on rabbits. Results: Results revealed that MET-loaded NC exhibited adequate EE% (62±3.2%), along with a PS of 222.2±11.25 nm which is ideal for the rapid penetration through vitreous gel, ZP of -23.9±2.67 mV and drug prolonged release behaviours from the in situ gel/film-forming system of MET-loaded NC followed by MET-loaded NC compared to free MET. Regarding in-vivo study results, MET-loaded NC in film forming solution revealed a significant reduction in IOP compared to free MET and plain NC in film forming solution. Conclusion: These findings indicated that the targeted co-delivery of MET using NC may offer a promising versatile therapy option for the treatment of vision threatening ocular diseasesItem ANALYSIS OF SELECTED VETERINARY DRUGS IN BIOLOGICAL FLUIDS AND/OR TISSUES(October university for modern sciences and arts, 2023) Hamed, Ahmed Gamal; Mohammed, Mariam Hani; Rezk, Mohamed Ashraf; Helmi, Rana AhmedAim Poultry are great sources of protein and other important nutrients for our body. So, poultry should be included as a part of a balanced diet. Residues of veterinary drugs in poultry meat have serious health effects on humans (e.g., antimicrobial resistance, carcinogenicity, and hypersensitivity), making controlling veterinary drug residues an important parameter in ensuring consumer protection. Hence, it is important to study the withdrawal time of these drugs in chicken tissues. Materials and Methods This work was performed to quantitatively determine a co-formulated veterinary formulation namely, Coccimix®, consisting of two sulfonamides (Sulfadimidine sodium and Sulfaquinoxaline sodium), in addition to Diaveridine, and Vitamin K3 in different tissues of broiler chickens including muscles (breast and thigh) and liver. The chickens were treated with the recommended dose of Coccimix® formulation. The analysis was done by solvent extraction for clean-up of samples from the tissue matrix, followed by HPLC-MS/MS method using Agilent C18 4.6*50 mm 2.7-Micron as a stationary phase and a mobile phase of acetonitrile: 0.1% formic acid in water (85: 15 v/v) with positive electrospray ionization. Results The proposed method was validated according to FDA guidance for bioanalytical method validation. The residues of the four components declined to reach levels lower than MRLs over a period of 7 days. Both sulfonamides and Diaveridine were found to be more persistent in liver tissues than muscle tissues. Conclusion It is recommended to monitor the withdrawal (elimination), especially for antibiotics, before marketing to ensure the hygienic suitability of broilers edibles for safe human consumptionItem Novel Therapy for Inflammatory Bowel Disease (RSPB2.5)(October University for Modern Sciences and Arts, 2023) Fawzy, Fawzy Ashraf; Ali, Nada Ahmed; Ahmed, Nourhan Abdelhameed; Ahmed, Omar AshrafIrritable bowel disease (IBD) is episodes of inflammation in the gastrointestinal tract, these episodes is caused by abnormal immune response to gut microflora. IBD is divided into two types: ulcerative colitis and crohn’s disease. The causes of IBD are increased permeability of intestinal barrier and imbalance in gut microbiota. Treatment known for IBD are anti-inflammatory drugs like corticosteroids and sulfasalazine, Immunosuppressive drugs like Imuran. Probiotics are foods or supplements that contain live microorganisms intended to maintain or improve normal microflora in the body. Therefore, the aim of study is investigation the efficacy of a novel strain of probiotics to be used as a potential treatment for IBD. While, the objectives are induction of inflammation on rats and mice, evaluation of novel therapeutic agent, and assessment of its anti-inflammatory activity histopathologically and by measuring inflammatory biomarkers. We worked on 3 colitis models: first model is BALB/c mice and colitis are induced by shigella toxin, second and third models are Wister albino rats and colitis is induced in one of them by acetic acid and in the other by dextran sulfate. We made protection with our novel therapy (lactobacillus) and commercial drug (enterogermina) or sulfasalazine. After the induction step, all mice and rats were scarified, and blood samples were collected to measure inflammatory biomarkers by ELISA and Western blotting, also intestine is isolated to be examined histopathologically. Histopathological examinations of acetic acid and Shigella models showed normal mucosa without alteration in the groups protected by lactobacillus, sulfasalazine and in control group, while the group protected by enterogermina showed mild edema. Elisa results proved that lactobacillus has anti-inflammatory activity as it showed inhibition in levels of inflammatory biomarkers after induction by acetic acid and shigella. In conclusion, lactobacillus showed more anti-inflammatory activity than enterogermina, so it is a promising drug in IBD treatment.Item The modulatory effects of kinase inhibitors on experimentally induced myocardial infarction(The modulatory effects of kinase inhibitors on experimentally induced myocardial infarction, 2023) Mohamed Ibrahim, Dina; Tarek, Gina; Hani Makram, Merihane; Kamal Yassin, YasminMyocardial infarction is the main leading cause of mortality world widely. In 2015, it is predicted that 7.4 million people died as a result of coronary heart disease. CVDs are predicted to kill 23.6 million individuals by 2030. Decreased or complete obstruction of the blood flow to part of the myocardium is the main cause of myocardial infarction. For the vast majority of those suffering from non-ST-segment elevation myocardial infarction (NSTEMI) as opposed to STsegment elevation myocardial infarction, it accounts for more than 15% of mortality each year. Myocardial infarction encompasses four different mechanisms which are: atherosclerosis development, deterioration of the formed fibrous plaque, thrombus formation and myocardial cell death. Tofacitinib is the drug under investigation in this study as it is suggested to have a cytoprotective effect against MI since it acts as inhibitor of the JAKs family which interferes with JAK/STAT signaling that occur following the inflammation that occur during myocardial infarction. This study aims to investigate both the relation between kinase inhibitors and myocardial infarction and the possible cytoprotective effect of Tofacitinib on experimentallyinduced myocardial infarction. In this recent study, 18 albino mice were used and randomly allocated into three groups. Group (A) was injected with saline only, Group (B) was injected with Isoprenaline HCl (100mg/kg) subcutaneously for induction of myocardial infarction and Group (C) was treated with Tofacitinib after receiving isoprenaline for 14 days. The parameters that were assessed were TNF-ὰ, cardiac troponin I, creatine kinase –MB, STAT-3, Lnc-Xist, NF-κB, mTor, IL-6 and there was histopathological examination. The results that were obtained from laboratory investigation of the previous parameters and histopathological examination proved the cytoprotective effect of Tofacitinib against MI as well as its ability to inhibit the inflammation and necrosis of the cardiomyctes that occur following the occurrence of MI. Therefore, Tofacitinib could be a used as a promising treatment and prophylaxis from myocardial infarctionItem Valorization of Citrus Wastes into Medicinally Active Anti-acne Pharmaceutical Preparation (RSPG 2.2)(October university for modern sciences and arts, 2023) Hashem, Adham Adel Abdelkader; Melek, Mariam Rashad Hezkial; Mahfouz, Nada Omar Mohamed; Mohamed, Naira AymanIn order to protect the planet and achieve sustainability, several approaches are suggested to reduce waste production and avoid too many different faces of pollution such as recycling and valorization. Valorization is to try to enhance the price, value, or status of products by organized and usually governmental action and our aim in this project is to valorize citrus waste and produce a pharmaceutical product for the treatment of acne. Citrus waste is estimated to be about15 million tons annually worldwide, including peels, seeds and fruit pulps. With Egypt producing around three million ton of citrus waste annually, valorization of citrus wastes into pharmaceutical product is considered a great tool to initiate and achieve sustainable production in the pharmaceutical field. Acne is an inflammatory skin disorder. The most common bacteria causing acne is Propionibacterium acne which triggers the inflammatory response in the skin causing swelling and redness, so the key route for treating acne is to inhibit the growth of this bacteria and treat inflammation. Many plants are known to have anti inflammatory such as Citrus fruits. Objective: The aim of the study is the valorization of the waste products of various Citrus fruits, exploring their chemical composition and evaluates their use in treatment of acne. Methods: The anti-inflammatory effects of the ethanolic extract of different citrus peels were tested in vitro on lipopolysaccharide(LPS)- induced inflammation in raw 264.7 cell model. The antimicrobial effect was evaluated against Propionibacterium acne and the metabolic profiling of the active extracts was performed using UPLC-MS/MS analysis. The ethanolic extract of Bitter orange is accordingly was chosen to be formulated as nano preparation to be tested in vivo. Results: It was found that bitter orange (Citrus aurantium), sweet orange (Citrus sinensis) and lemon (Citrus limon) peels extracts were the most effective as they inhibit NO and PEG2 production. Also, RT-PCR showed that the three extracts had down regulated the gene expression of COX2 and iNOS. These results were confirmed by the Western blotting assay. Thus, the three extracts were chosen for further study of the antibacterial activity against Propionibacterium acne, where the bitter orange extract showed the lowest MIC 1.875 mg/ml, while lemon and orange extracts had higher MIC (30 and 15 mg/ml). The metabolic profiling of bitter orange extract showed the VI identification of many flavonoids such as hesperidin and vicenin II. Bitter orange in-vivo testing shows promising results where the medicated nano-micelles in the mice of group four showed comparable activity to the Erythromycin. Conclusion: Bitter Orange (Citrus aurantium) shows promising in-vivo anti acne activityItem The pharmacological study of tetracycline uses in an infected diabetic wound healing in rat model RSPHO 2.4(October University for Modern Sciences and Arts, 2024) Hassanein, Alaa Hamdy Mohamed; Helal, Mariam Wael Mohamed Ahmed; Mansy, Mayar Mostafa Ali; Hiekal, Reem Hisham Ahmed1. Abstract Since the skin is the part of the human body that encounters the world most directly, it is particularly vulnerable to injury. When injuries take place, the skin will lose its function and its integrity, which may result in a life-threatening situation. Wounds may occur due to various causes that affect the wound healing normal physiological process which consists of four main stages: homeostasis, inflammation, proliferation and tissue remodelling. However; patients with diabetes mellitus (DM) suffer from wound healing impairment. DM has many drawbacks which influence the four stages of the healing process causing a delay in the healing, or proceeding to chronic wounds. Moreover, wounds may get infected due to bacterial colonization; thus antibiotics are used to defend the body against the bacterial infections such as tetracycline. Diabetic rat models with excisional wounding inoculated with staphylococcus aureus bacteria are used in the experiment, divided into 3 groups: control, standard tetracycline (suspension) and Sobulous patch containing tetracycline groups. Rats are then sacrificed to be tested physiologically and histologically in order to determine the difference between pharmacological effects of the market available tetracycline and the novel dosage form in the healing process and its effect on the collagen regeneration. The biochemical markers TNF-α, IL-1B, IL-10 are measured using ELIZA, while the MMP-9 is measured through PCR. The results are promising as the tetracycline decreases the pro-inflammatory mediators and improve the anti-inflammatory mediators resulting in improved re-epithelization, granulation tissue formation, and accelerated wound closure. The aim of the study is to establish a wound healing model with a novel dosage form to increase the available market options for the targeted patients.Item Advanced Extraction Protocol(s) for Analysis of Selected Antiviral Drug(s) (Entecavir).(RSPAC2.4)(October University for Modern Sciences and Arts, 2024) Yaser, Ahmed; Alaaeldin, Hosam; Saber, Mahetab Osama; Eissa, Mennaallah MohamedAbstract : Entecavir (ENT) which have a structure that contain a guanosine nucleoside analog, is used as antiviral medication because of it is selectivity toward the chronic hepatitis B virus (HBV) that will inhibit it. A shift or earlier combination of lamivudine with ENT will result less resistance, comparable safety and more successful viral suppression compared to monotherapy, according to a clinical guideline. Additionally, there are several ways that ENT might degrade. Therefore, it is crucial to prepare certain materials to extracting the ENT. Molecular imprinting polymers (MIP) were created for this use through precipitation polymerization using the non-covalent method. Transmission electron microscopy (TEM) was used to characterise the polymer. The TEM results showed that polymer particles have a spherical shape and homogeneous size. Studies on ENT degradation by oxidation, basic and acidic force were performed. The stated drug, including its co-administered medication (lamivudine), and its breakdown products, were extracted from spiked plasma of human using the MIP. Using a recently developed HPLC technique that used a C18 column, gradient elution from the mobile phase made up of 0.1 percent phosphoric acid (H3PO4) in water : methanol with such flow rate of 1 ml/min, and UV absorbance around 254 nm, all eluents were examined. Forced degradation experiments were used to demonstrate the method's specificity as well as stability-indicating capacity. When MIP's selectivity was compared to that of ENT itself and degradation products, the difference was highly noticeable. Additionally, ENT was effectively extracted out from spiked plasma of human using MIP like an extractant, with such recovery rate of 88.32%.Item STUDYING THE REGULATORY ROLE OF NON-CODING RNAS AS NOVEL DIAGNOSTIC AND THERAPEUTIC TARGETS IN PULMONARY DISEASES. (RSPB2.7)(October University for Modern Sciences and Arts, 2024) Hassan Eldeeb, Mazen Essam; Roshdy, Maram Mohsen Mostafa; Rashad, Doaa Adel; Abdelsallam, Nadine HussienABSTRACT Objectives: Investigation of the pulmonary protective effect of miRNA inhibitor against LPS-induced Acute respiratory distress syndrome (ARDS), the predictable serious complication of COVID19 by targeting ACE2/ Angiotensin pathway. Materials and methods: BALB/c Male mice were randomized into 3 groups (n= 6 mice/group); the first group is the normal control group who received normal saline, the second group is the induction group who got injected with lipopolysaccharide (LPS) intraperitoneally, while the third group is the treatment group who got injected with miR-200 inhibitor intraperitoneally two hours before the LPS injection. Lung samples were collected. The right lungs will be preserved at -80 ºC for the biochemical analysis for miR-200 and ACE2, while the left lungs will be fixed in 10% formalin solution for the histopathological investigations for of NF-κB, TNF-α and IL-6. Results: The treated mice showed marked improvement in ACE2 levels with normal histological structure of lung tissue. There was also a downregulation of miR-200 and Ang 2 levels and marked elevation of Ang 1-7 levels Conclusion: The upregulation of miRNA-200 is considered an early potential noninvasive biomarker, while miRNA-200 inhibitor is considered to be a therapeutic agent for ARDS.Item Evaluation of the anti-inflammatory effect of Prunes persica extract in rats(October University for Modern Sciences and Arts, 2024) Nour, Tarek; Mahmoud, Moustafa; Atef, Sara; Ayman, MarinaABSTRACT The aim of this study to evaluate the anti-inflammatory effect and the mechanism of action of Prunus persica which belongs to the family Rosacea, It is widely cultivated in China and very rich in phytochemicals like phenolic compounds, carotenoids, vitamins, volatiles and organic acids. In this study, rats are injected by carrageenan which causes inflammation through stimulation of inflammatory mediators such as histamine and bradykinins to be released, then treated with the extract of Prunus persica and assay the inflammatory markers .Inflammation is the biological response to a number of factors including pathogenic agents, damaged cells and toxic substances in the immune system. These factors may cause acute or chronic inflammatory responses, potentially leading to tissue damage. Inflammation at tissue level is characterized by redness, swelling, heat, pain, tissue loss and the resulting immune, vascular and inflammatory response from local cells to infection or injury. The inflammatory cells and the trigger of inflammatory signaling pathways are triggered by infectious and non-infectious agents and cell damage .The inflammatory markers such as iNOS (Induced Nitric oxide synthase ) is one of the main inflammatory mediators involved in both inflammation and angiogenesis NO is produced due to cytokine-induced expression of (iNOS) inducible nitric oxide synthase which largely involved in the pathophysiology of inflammation , COX-2 which is a highly inducible in the response of pro-inflammatory stimuli, Selective inhibition of COX-2 might lead to more effective anti-inflammatory medicines that are safer than current NSAIDs. NSAIDs (non-steroidal anti-inflammatory drugs) are medications that are commonly used to alleviate pain, decrease inflammation, and lower a high fever that work by inhibiting the cyclooxygenase (COX) enzyme and, as a result, prostaglandins at the site of inflammation. Unfortunately, inhibiting gastrointestinal or renal prostaglandins is linked with mechanism-based toxicities, limiting the use of these otherwise effective and powerful medicines.COX-2 is strongly inducible in the reaction to pro-inflammatory stimuli, cytokines and mitogens, so it has typically been equipped to play a significant role in the inflammatory release of PGs. COX-2 is constitutively found in the whole forebrain and in discrete neuronal groups of cortex and hippocampus, it is largely responsible for causing inflammation. IL-6 has been linked to a variety of inflammatory-related chronic diseases. Monocytes and macrophages produce IL-6 in response to inflammatory cytokines such as IL-11 and tumour necrosis factor (TNF)-alpha. The IL-6 receptor is found on RSBP2.2 Evaluation of the anti-inflammatory Effect of Prunes Persica Extract in Rats IX resting T-lymphocytes, activated B-cells, and myeloid and hepatic cell lines. Inflammation will be induced by a single dose of carrageenan in twenty four rats which will be randomly divided into four groups: Group 1: six normal rats, Group 2: six rats have inflammation .Group 3: six rats which have inflammation and control with Diclofenac, Group 4: 6 six rats which have inflammation will be injected with 100mg/dl of Prunus persica extract. When the paw edema is examined in the rat the standard group which have inflammation and treated with Diclofenac and Test which they have inflammation and treated with Purnus Persica extract are compared the standard and test to injured in 0 hour , 1 hour , 2 hours , 3 hours and 4 hours they are significantly decreased compared with injured. In the Histopathological examination, Tissue section of rat paw skin treated by diclofenac sodium showing marked reduction of inflammatory reaction score 1 and tissue section of rat paw skin treated with a topical gel formulation containing Prunus Persica extract shows a low level of inflammation. When Interleukin 6 and Nitric oxide levels are assessed in Diclofenac (Voltaren) and extract-treated rats, they are substantially lower than in carrageenan-treated rats..Item Exploring the Immunostimulant Activity of Selected Nuts in Relation to their Metabolic Profile RSPG 2.1(October University for Modern Sciences and Arts, 2024) Naguib, George Zaky; Ahmed, Zeinab Mohammed; Mahmoud, Amira Mohamed; Mostafa, Radwa MohamedABSTRACT Immune system is the potent army that defends our body against various infections and diseases through innate and adaptive immunity. Immunostimulant are those substances that activate the immune system by triggering stimulation or rising activity of any of its components. Nowadays, COVID-19 is a pandemic that lasts with no specific treatment and maximizing the capability of immune system defense is one of the routes for its management. Herbal medicine is one of the essential sources for enhancing immunity because of their affordability, availability, minor side effects and people preference. Hazelnut, Walnut, almond and peanut are from the most widespread edible nuts that are rich in fats, fibers, vitamins, proteins and minerals. Our aim is to determine the shells and edible nut in vitro immunostimulant activity as well as metabolomics profiling of the four nut extracts. 500 grams of each of nuts and shells were powdered then defatted and extracted by using n-hexane and ethanol (95%) followed by evaporation of the pooled extracts under reduced pressure using the rotary evaporator in a temperature not exceeding 60˚C followed by measurement of yield of each of hexane and ethanol extracts. For nuts, total phenolic were found to be in Peanut Almond, Hazelnut and Walnut of 13.4538, 17.8237, 9.7978, and 17.6237 respectively and for shells, 240.0753, 21.4559, 159.6989, and 328.0860 respectively. The antioxidant activity of the shells and nuts was measured by: DPPH, and ABTS assays through measuring inhibition percentage as well as FRAP by measuring absorbance which have been indicated the significant antioxidant capability especially walnut peanut shells. The toxicity of each of nuts’ and shells’ extracts has been assessed though MTT viability test which indicated their safety on the differentiated THP-1. Addition of extracts to LPS-stimulated macrophages especially peanut and walnut shells have downregulated the expression of AP-1 and in turn decreased TNF-α and IL-8 production. On the other hand, NF- κB contributed to the downregulation of iNOS and COX-2. UPLC-MS metabolic profiling of the four nuts shells and nuts has been done detecting around 128 metabolites.Item Genotypic study of the association between CRISPR/Cas system and antimicrobial resistance in Klebsiella pneumoniae (RSPM2.1)(October University for Modern Sciences and Arts, 2024) Nessim, Anton; Eid, Gamal Adel; Mohamed, Hassan; kamal, Mahmoud AhmedABSTRACT CRISPR-Cas system is a recently discovered genomic engineering tool with the potential for knocking-out and knocking-in sequence-specific DNA targets. CRISPR is an abbreviation for Clustered regularly interspaced short palindromic repeat while Cas stands for CRISPR-associated enzymes. CRISPR/Cas has been originally known as a bacterial adaptive immune system utilized by host microbes for fighting foreign DNA invaders such as phages and plasmids. It has also been linked to higher susceptibility to antimicrobial agents in Enterococcus species as well as in Escherichia coli. Recent reports extending such findings to Klebsiella pneumoniae has been also published. This presents CRISPR/Cas as a potential solution for antimicrobial resistance. K. pneumoniae is a highly problematic species known for multidrug resistance and hypervirulence of some strains. The current study aimed at characterizing CRISPR/Cas systems in K. pneumoniae and their potential impact on acquisition of antimicrobial resistance. For this purpose, a collection of 46 K. pneumoniae isolates were tested for antimicrobial susceptibility as well as harboring CRISPR/Cas genes. Susceptibility profiles were analyzed using disk diffusion test and CRISPR/Cas genes were amplified using Polymerase Chain Reaction (PCR). The analysis revealed susceptibility to higher number of antimicrobials among the group of isolates carrying Cas genes compared to the Cas-negative isolates. As only five isolates were found to carry Cas genes, such findings were not considered as conclusive and a large scale bioinformatic analysis was done on a sample of K. pneumoniae genomes retrieved from the NCBI database. A total of 337 genomes were scanned for CRISPR/Cas genes which were identified in only 20% of the studied genomes. MLST analysis of the CRISPR/Cas positive isolates revealed that CRISPR/Cas systems were disseminated in different sequence types most commonly in ST23, a well-known multidrug resistant hypervirulent ST. Surprisingly, CRISPR/Cas-positive strains were found to carry larger number of resistance plasmids compared to others. In conclusion, CRISPR/Cas systems are disseminated with low prevalence in various sequence types and correlation to antimicrobial susceptibility is questionable. Further studies are required to investigate the role of CRISPR/Cas in acquisition of resistance by K. pneumoniae. A larger number of isolates should be phenotypically and genotypically analyzed.Item MOLECULAR MECHANISMS UNDERLYING IMMUNOMODULATORY PDT OF DERMATOLOGICAL MELANOMA (RS 502) (RSPB2.1)(October University for Modern Sciences and Arts, 2024) Tarek, Hadi; Nour, Asaad; Mostafa, Asmaa; Ahmed, Ibrahim1. ABSTRACT Increased exposure to the ultra violet radiation (UVR) from the sun, as well as lack of proper consciousness, has led to fatal malignancy with an increased mortality rate in the last ten years due to skin melanoma development, especially in people with fair and light skin. The current treatment scheme for melanoma includes chemotherapy, radiotherapy and surgery. Tumorigenesis is complex and dynamic at three levels: initiation, progression, metastasis. In addition, there is a tight connection with the tumors, the tumor microenvironment (TME) and the extracellular matrix, in each level (ECM). Photodynamic therapy (PDT) is a minimum-invasive therapy, which combines the use of a photosensitizer (PS) with laser exposure. When the laser beam of a specific wavelength is exposed to photosensitizers, it produces reactive oxygen free radicals which can kill the exposed cells in the vicinity. A specific wavelength enables each photosensitizer to produce its action. This wavelength can determine the extent to which the light can pass through the body. Moreover, PDT is associated with immune-stimulation that inhibits cancer progression through apoptosis and tumor cell necrotization. Immune system stimulation can be detected with several biomarkers like IL10, IL12, TGF-ß and TNF-α. This work aims at studying some of the genetic markers involved in the molecular mechanisms of PDT mediated immunomodulatory treatment of skin melanoma. Our target is to explore the relation between PDT as a recent efficient method for treating oncogenic tumors and the role of a tumor microenvironment in monitoring the development of skin cancer.Item Pharmacogenomics role and personalized medicine in osteosarcoma patients (RSPL2.3)(October University for Modern Sciences and Arts, 2024) Mohamed, Fatma Ahmed; Mostafa, Mariam Gamal; Zenhom, Rana Elamir; Rabea, Aliaa MuhammedAbstract The presence of malignant mesenchymal cells producing osteoid or immature bone defines Osteosarcoma (OS). It is the most common primary bone tumor in children and adolescents, accounting for approximately 5% of childhood tumors. OS has a bimodal age distribution, with peaks in adolescence and older adults. The metaphysis of long tubular bones is the most common site of OS, while it is rare in the spine, pelvis, and sacrum. In the 1970s, amputation was the standard treatment for OS, but the survival rate was less than 20%. The combined limb salvage treatment and neoadjuvant chemotherapy in the 1980s increased the survival rate to nearly 80%, but there has not been any improvement since then. Long noncoding RNAs (LncRNAs) are 200 nt length RNA molecules that can regulate gene expression. HOTAIR, a well-known LncRNA, is involved in the pathogenesis and progression of multiple tumors, but little is known about the role of gene polymorphisms in the carcinogenesis of OS. Understanding the different possible causes of pathogenesis could aid in improving OS treatment and survival. Aim: the study aims to Investigate the role of Single nucleotide polymorphism in Hox transcript anti- sense RNA (HOTAIR) in Osteosarcoma Pediatric Patients in the Egyptian population, two single nucleotide polymorphisms were selected in line with literatures supporting the association of the selected single nucleotide polymorphisms with osteosarcoma, the selected single nucleotide polymorphisms were rs920778 and rs7958904. Methods: 90 blood samples were collected retrospectively obtained from the biobank, followed by DNA extraction of blood samples using QIAamp kits prior to QPCR amplification and genotyping performed. Finally, data analysis was conducted using the R program and SPSS. Results : 90 samples were collected, however, only the clinical data of 50 patients was obtained, our findings revealed the existence of SNP rs920778 in all the patients, where 22 patients 44% harbored Heterozygous Allele 1/Allele 2 , 12 patients 24% harbored Homozygous Allele 1/Allele 1 and 16 patients 32% harbored Homozygous Allele 2/Allele 2 , while in SNP rs7958904 23 patients , 46 % don’t harbor the SNP at all while 13 patients ,26% harbored the homozygous Allele2/Allele2, and 14, 28 % harbored the heterozygous Allele1/Allele2, Patients’ clinical characteristics were associated with SNP rs920778 and SNP rs7958904 revealing interindividual variation between patientsItem Melon By-products and their Biopotential (RSPG2.6)(October University for Modern Sciences and Arts, 2024) Fathy, Aya Hamdy; Mohamed, Kholoud Mohamed; Ahmed, Laila Mohamed; Abd el-fatah, Menntallah AshrafABSTRACT Wastes of edible fruits represent a wealth with their phytoconstituents, They can be used in dietary supplements and skin care. In this project, Cucumis melo var.cantalopensis (Cucurbitaceae) was investigated for its phytoconstituents in correlation to the antioxidant and anti-aging effects The Wastes of c.melo var.cantalopensis (seeds and peels) were extracted with hexane to be analyzed for its fatty acids, sterols and hydrocarbons using Gas chromatography/mass spectroscopy (GC/MS). The main unsaturated fatty acid was methyl linoleate while the main saturated fatty acid was methyl palmitate, furthermore, the predominant hydrocarbon was octacosane. Vitamin content estimation revealed a high level of vitamins C and A in peels which were 16.52 and 24.03 mg/g respectively. Both peel and seed have a high level of minerals hence peel is rich in Ca, Mg and Fe and their amounts were 0.823, 0.432 and 192.873 mg/g, respectively on the other hand seed are rich in Se, Cu and Zn which their amounts were 4.410, 11.079 and 40.902 mg/g respectively. Phenolic content was estimated with Folin–Ciocalteu method and its value was 11.1mg/g while flavonoid content was 6. 5 mg/g. Anti-oxidant activity was tested using three assays which were DPPH, FRAP and ABTS. In DPPH free radical scavenging activity, extracts of C.melo showed that percentage inhibition of the methanolic extract of peel is 5.63 +/- 0.07 and seed is 11.68 +/- 1.20, in FRAP, results of peels and seeds ranges were 151.7 +/- 15.8 and 44.81+/- 6.44 microM TE/mg extract, respectively and in ABTS the range of values of peel was 160.94 +/- 12.85 and for seed was 19.63 +/- 2.71 micromolar TE/mg extract. Comparing the inhibitory activity of our extract and oil on tyrosinase, collagenase and elastase enzymes as representatives for anti-aging activity, it is noted that methanolic extract of peel has better inhibitory activity on the 3 enzymes more than oil and those results are comparable with the standards of the positive control. The percentage inhibition of anti-elastase in methanolic extract was 86.12 +/- 1.32 and in oil was 87.92 +/- 2.21, the percentage inhibition of anti-collagenase in methanolic extract and oil were 61 +/- 2.1, 75.23 +/- 1.65 respectively and the percentage inhibition of anti-tyrosinase in methanolic extract was 61.03 +/- 1.85 and in oil was 74.12 +/- 1.50. The C.melo represented a promising candidate in pharmaceutical skin care market.