Master In Chemistry

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    First molecular detection and genotyping of Neospora caninum in naturally infected cattle and sheep in Palestine
    (Al-Quds University, 2024-05-05) Heba suleiman salem alfarajeen; هبة سليمان سالم الفراجين
    Neosporosis has become one of the most common diseases causing abortion in dairy cattle globally. The clinical signs of Neospora caninum have been reported in sheep, goats, deer, and horses. Fetal abortion induced by N. caninum is a common reproductive problem that causes significant economic loss in cattle and sheep husbandry. This study aimed to detect and identify N. caninum in intermediate hosts such as cattle and sheep from Palestine using polymerase chain reaction (PCR) followed by DNA Sanger sequencing and to study the genetic diversity of the study samples, targeting the MS10 microsatellite, using phylogenetic analysis. A total of 124 brain tissue samples were collected from 106 (85%) cattle and 18 (15%) sheep from a slaughterhouse in Jericho-Palestine. The PCR technique was used to identify the Neospora DNA in the brain samples based on the Nc-5 gene. N. caninum was detected in the brain samples; out of 124 samples, 30 (24.19%) samples were positive for N. caninum. The frequency of N. caninum in cattle (25.47%) was higher than that of sheep (16.66%). The infection caused by N. caninum was confirmed by DNA Sanger sequencing of ( 13 ) random positive samples. BLAST analyses of the Nc-5 gene revealed more than 92% to 99% matching with N. caninum sequences available in GenBank. Microsatellite MS10 repeats were used to study the genetic diversity of the Palestinian isolates. The microsatellite genotyping was done by next-generation sequencing (NGS). It was applied to 30 positive samples, and successful results were achieved in 15 samples. Our study sequences displayed 100% similarity with N. caninum isolated from dogs in Liverpool and 99.34% similarity with N. caninum isolated from cattle in Japan. The phylogenetic analysis of 15 N. caninum samples from Palestine and 83 microsatellite (MS10) repeats obtained from different countries and different hosts showed that N. caninum isolates are genetically diverse and distributed into v two main clusters. Locally, the Palestinian isolates are distributed in two clusters and share the same repeats with samples from Asia and Europe. The present study provided the first estimate of the frequency of N. caninum in cattle and sheep from Palestine. Additionally, it was also the first time for the investigation of the phylogenetic analysis of N. caninum based on microsatellite markers.
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    APOE Gene variants and Risk of Hyperlipidemia in Palestinian Type 2 Diabetes Mellitus Patients
    (Al-Quds University, 2021-01-16) Manal Mohammed Saleh Ghattas; منال محمد صالح غطاس
    Diabetes is considered as one of the most popular metabolic diseases worldwide. Type 2 is the most common by which it affects 90% of all diabetic patients in the world. The problem with the disease is that insulin is not doing its job or it is not secreted at all. This elevates the sugar level in the bloodstream, as it is not entering the human cells. There are several contributing agents that participate in the development of T2DM, they could be environmental or genetic or both. Many genes have been shown to be involved in increasing the risk of T2DM. One of these genes is APOE gene which is responsible for producing apolipoprotein E that is important in lipid metabolism. The study is a cross sectional that was done in the period from January to April 2019. It consists of T2DM Palestinian subjects divided into two groups; lipidemic (case) and non-lipidemic (control). A total of 204 diabetic subjects were included in this study; 96 participants were lipidemic and 108 were non-lipidemic. All patients aged over 50 years and admitted in Ramallah hospital/Palestine. The lipid profile was performed during the hospital admission examination. Two SNPs within the exon 4 of the APOE gene and three SNPs in the APOE promoter region were identified by amplicon based next generation sequencing (NGS) and it is the first to be held on Palestinian population. The frequencies of these polymorphisms were assessed in the studied population. Further, the associations of APOE genotype and promoter SNPs with risk of hyperlipidemia in T2DM patients were investigated. Our results showed that the frequency of E3 allele was the highest (79.6%) among other alleles; E3/E3 genotype was also found to be highest in all T2DM subjects with a frequency of 78.1%. Our study population was stratified according to lipid status, the obtained results showed no statistical differences (P>0.05) of APOE genotypes frequency (APOE polymorphisms and promoter SNPs) among lipidemic and non lipidemic groups. Further, Logistic regression analysis adjusted for age, gender and BMI showed no association between APOE genotypes (epsilon and promoter polymorphisms) and risk of dyslipidemia. No significant association was observed between APOE genotypes and serum cholesterol, TG, HDL-C and LDL levels. The possible synergistic effect between the promoter SNPs and the common APOE polymorphism was also investigated. We found no significant differences between lipidemic and non lipidemic groups after stratification of the promoter SNPs with the APOE genotypes. In conclusion, association between known SNPs and diseases are varied among different ethnic groups. Our study showed no association between APOE genotypes and hyperlipidemia in type 2 diabetic patients in the Palestinian population. Further studies should be done on larger sample to confirm the study results.
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    synthesis and characterization of novel polymer based on hydroxamic acid
    (Al-Quds University, 2021-08-14) Rawdah Ma'n Eid Sunoqrot; روضه معن عيد سنقرط
    Water-soluble polymers based on Hydroxamic acids are becoming increasingly important in several applications for their impressive chelating ability with heavy metals.In this study Acrylo- hydroxamic acid monomer was prepared from the reaction of n-butyl acrylate with hydroxylamine hydrochloride using potassium hydroxide as a base in methanol. The linear polymer was prepared at room temperature in degassed distilled water using free-radical initiators, potassium persulfate (K2S2O8), and sodium metabisulfite (Na2S2O5) under inert gas (N2) conditions. A cross-linked polymer was prepared by using potassium persulfate (K2S2O8) as a free radical initiator and N, N-Methylenbis (acrylamide) as a cross-linking agent at 70 C. The polymers produced in this work have a high weight average molecular weight (Mw). The average weight of linear and cross-linked polymers were determined by light scattering and they were 488000, 431000 gm/mol respectively
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    Phase behavior of Basil extractions and their favorable attributes
    (Al-Quds University, 2021-12-21) Hadba Mousa Issa Tarayrah; هدبه موسى عيسى طرايره
    Nowadays there is a great focus on developing medicines, and searching for plant sources in order to reduce side effects. Moreover, research into new techniques to increase the effectiveness of the active substance obtained.Natural products extraction has played a key role in the discovery of leads for the development ofnew drugs. Basil plant was selected for our study, which has more than 200 chemical compounds. In this study, basil leaves and seeds were extracted through the use of the Soxhlet extraction apparatus, This extraction method that we developed and modified in our laboratory was able to extract almost all active compounds from this plant.. The in vitro susceptibility for basil extracts was determined against antibacterial activity by agar disc diffusion method, minimum inhibitory concentration and minimum bactericidal concentration for Eschericha coli, Staphylococcus aureus, Klebsiella pneumonia, Enterobacter sp, and Pseudomonas aeruginosain the presence of positive control (Gentamicin (10 μg/disc) and Penicillin 10 unit ). The findings demonstrated that ethanol extract seeds and leaves exhibit antibacterial action against the majority of bacterial strains. The highest zone of inhibition for seeds extract against Enterobacter sp. was 10 mm at 100 mg/ml concentration, whereas the highest zone of inhibition for leaves extract against Enterobacter sp. was 25 mm at 100 mg/ml concentration as diffusion (spread).A strain of Staphylococcus.aureus has an MIC of 62.5 μg/ml, represents the highest efficacy of the extracts against bacteria strains. The extracts of leaves and seeds were evaluated for their antioxidant activity (AA) which was determined by using a modified method of the assay of ferric reducing/antioxidant power (FRAP), and the total phenolic content (TPC) by the Folin-Ciocalteu assay. All the analysis was made with the use of a UV-Visible spectrophotometer. Leaves and seeds showed strong antioxidant ability and high phenolic content.The extraction of a seeds part of the plant has 74 ±1.4 mg/gantioxidant activity, comparing this value with those obtained by leaves part 40.4 ±0.8 mg/g. while the total phenolic content (TPC) for Basil seeds (58.2 ±0.9 mg/g ) while that of Basil leaves (51 ± 2.4 mg/g). Moreover, analysis of their flavonoid content (TFC) by using HPLC chromatography at 254 nm. flavonoid compounds were detected in 6.60 minutes for Basil seeds while detected in 6.60 minutes for Basil leaves. Furthermore, total flavonoids content (TFC) was measured by using the aluminium chloride spectrophotometric method. The total flavonoids content (TFC) in leaves (9.0 ± 1.5 mg/g) while for seeds (34.2 ± 3.6 mg/g). The potential in vitro antimalarial activity of different extracts of the Palestinian Basil herb (Ocimum) was done in different solvents (ethanol 99.5%, ethanol 35%, ethyl acetate, petroleum ether and water) using a semi-quantitative assay method it was found that β-hematin in either one of these solvents is not prevented.This result shown for the first time in this study , rules out one mechanism proposed to explain the plant’s antimalarial activitythat is inhibition of beta-hematin formation. This study aims to formulate phase behavior using leaves and seeds extracts from Basil, Tween 20 as a surfactant and Co-surfactant also was used with absolute ethanol.This investigation is the first report on the formation and production of pseudo ternary phase behaviourfor Palestinian Basil (leaves and seeds ) extracts. In this research, it was found that Palestinian basil has the capability to the formation of microemulsion in vitro systems. Microemulsion technology was used to produce a medical cream. The formula is moisturizing, smoothing, good smell, absorb by skin (don’t leave film), not irritation pH= 5-5.5, good color, easily spread (not too viscous, apply easily and quickly), and anti-aging, antierythmic, and depigmenting effects when applied topically. Based on these findings, it is concluded that Basil constitutes a natural source of potent antioxidants, antibacterial, anti-aging, antierythmic, and depigmenting effects. that may prevent many diseases and could be potentially used in cosmetics, and pharmaceutical products. في الوقت الحاضر هناك تركيز كبير على تطوير الأدوية والبحث عن مصادر نباتية لتقليل الآثار الجانبية. بالإضافة الى البحث في تقنيات جديدة لزيادة فعالية المادة الفعالة التي يتم الحصول عليها. اناستخلاص المنتجات الطبيعيةلعبت دورًا رئيسيًا في تطويرأدوية جديدة. حيث تم اختيار نبات الريحان في هذه الدراسة لإحتوائه على أكثر من 200 مركب كيميائي. في هذه الدراسة تم استخلاص أوراق الريحان والبذور من خلال استخدام جهاز الاستخلاص (Soxhlet), هذه الطريقة قادرة على استخلاص جميع المركبات النشطة تقريبًا من هذا النبات. وقمنا بتطويرها وتعديلها مخبريا. تمت دراسة الفعالية المضادة للبكتيريا باستخدام طريقة القرص, تم تحديد اقل تركيز من الماده الفعاله يمكن ان يمنع نمو الكائنات الدقيقه او يقضي عليها كليا (MBC,MIC ) لكل من Eschericha coli ، و Staphylococcus aureus ، و Klebsiella pneumonia ، و Enterobactersp ، و Pseudomonas aeruginosa في وجود تحكم إيجابي ( (Gentamicin 10 ميكروغرام / قرص) ووحدة بنسلين 10). أظهرت الدراسة أن كلا من مستخلصات البذور والأوراق بالإيثانول لها نشاط مضاد للجراثيم على معظم السلالات البكتيرية. فقد كان أعلى تركيز لتثبيط مستخلص البذور هي 10 ملغم/ مل بتركيز 100 ملغم/ مل ضد Enterobactersp. بينما تبلغ أعلى تركيز لتثبيط مستخلص الأوراق 18 ملغم/ مل بتركيز 12.5 ملغم/ مل ضد Enterobactersp و Staphylococcusaureus على شكل انتشار. تم تحليل مستخلص البذور والأوراق باستخدام طرق الفحص القياسية, حيث تم فحص نشاطها المضاد للأكسدة (FRAP) ومحتوى الفينول (TPC) بطريقة فولين Folin-Ciocalteu. وقد تم التحليل باستخدام جهاز UV-Visible spectrophotometer. وقد أظهرت الدراسة ان بذور وأوراق الريحان تحتوي على قدر عالي من مضادات الأكسدة ومحتوى الفينول. حيث بلغ محتوى نسبة مضادات الأكسده في البذور على 74 ±1.4ملغم/غم, بينما في الاوراق 40.4 ±0.8 ملغم/غم. أما نسبة المحتوى الفينولي (TPC) في البذور 58.2 ±0.9 ملغم/ مل كانت أعلى من محتوى أوراق الريحان 51 ± 2.4 ملغم/ مل. بالإضافة الى ذلك, تم تحليل محتوى الفلافونويد (TFC) باستخدام HPLC عند 254 نانومتر. تم اكتشاف مركبات الفلافونويد في 6.679 دقيقة لبذور الريحان بينما تم اكتشافها في 6.682 دقيقة لأوراق الريحان. أما نسبة محتوى الفلافونويد (TFC) فتم قياسها بطريقة الفحص اللونية Aluminium chloride. حيث كانت بالبذور 34.2 ± 3.6 ملغم/ مل, بينما في الأوراق 9.0 ± 1.5 ملغم/ مل. تم أيضًا فحص فعاليتها المضادة للملاريا وقيامه بمنع تكوين البيتا هيماتين في المختبر تحت ظروف فيزيائية وكيميائية. لم يلاحظ أي نشاط ضد الأنشطة المضادة للملاريا لأجزاء مختلفة من مستخلصات الريحان (الأوراق والبذور) في المذيبات المختلفة. تهدف هذه الدراسة إلى صياغة سلوك مرحلة المستحلبات الدقيقة باستخدام مستخلصات الريحان (الأوراق والبذور) ومنشط السطوح(Tween 20)كمادة خافضة للتوتر السطحي ومساعد منشط السطوح (co-surfactant)وهو الايثانول. هذه الدراسة هي أول تقرير عن تكوين وإنتاج سلوك المرحلة الثلاثية لمستخلصات الريحان الفلسطيني (الأوراق والبذور). في هذا البحث، وجد أن الريحان الفلسطيني لديه القدرة على تكوين مستحلب المايكرو(microemulsion) في الأنظمة المخبرية. تم استخدام تقنية Microemulsion لإنتاج كريم طبي مقاوم للشيخوخة والتأثيرات المضادة للطفح الجلدي ومزيل للتصبغات عند تطبيقها موضعيًا. يمتاز بالترطيب والتنعيم والرائحة الطيبة كما انه يمتص بسهولة بواسطة الجلد ولا يحدث تهيجا pH = 5-5.5، كما ان لونه جيدا وينتشر بسهولة (ليس لزجًا ويطبق بسهولة وبسرعة). بناءً على هذه النتائج، استنتج أن الريحان يشكل مصدرًا طبيعيًا لمضادات الأكسدة القوية، ومضادات الجراثيم، ومضادة للشيخوخة، ومضادة للطفح الجلدي، ومزيلات التصبغ. التي قد تمنع العديد من الأمراض ويمكن استخدامها في مستحضرات التجميل والمنتجات الصيدلانية.
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    Preparation of semi-solid dosage forms containing Psidium guava leaves extract as antimicrobial preservative
    (Al-Quds University, 2021-08-21) Mohammad Daud Abdallah Atieh; محمد داود عبدالله عطية
    As it is known, guava leaves extract has several medicinal characteristics, such as antimicrobial, antioxidant, anti-cancer and anti-inflammatory. Several studies have been proposed to use guava leaves extract in several preparations. However, applying guava leaves extract as a preservative in a pharmaceutical product has not been studied in previously. In the present work, we try to use a natural preservative instead of chemical (methyl and propyl paraben) since many studies were warning from their effect and that it must be used at limited concentration. Guava leaves were extracted with ethanol 95.5 % as extraction solvent. The percentage yield was 20% which was acceptable. This extract was tested by HPLC; several phenolic compounds were detected. To determine the ability to use this extraction powder as a preservative, antimicrobial effectiveness test (preservative efficacy test) was conducted for extracted powder in purified water at different concentrations (0.5, 2.5, 5, 7.5, 9.5, 10) % w/w, against three bacteria, one gram-positive (S. aureus), two gram-negative (P. aeruginosa, E. coli) and two fungi: one yeast (Candida albicans) and one mold (Aspergillus (Niger) brasiliensis). The results showed that the only concentration that passed the test was 10%; this concentration was used in semi-solid pharmaceutical products. For this purpose, six different preparations (Ketoconazole shampoo, Clotrimazole Cream, Permethrin Cream, Gentamycin Cream, Ibuprofen gel, Indomethacin emulgel) were used. Chemical preservative was replaced by natural one (guava leaves extract powder) to serve as a preservative. Only three out of all six preparations (Ketoconazole shampoo, Clotrimazole Cream, Ibuprofen gel,) passed preservative efficacy test. This passed preparations were fully checked-up by chemical (drug content,) physical (odor, physical appearance) and biological test (Direct transfer test, Total count test) for three months at accelerated condition. The results confirmed that pharmaceutical preparations were stable and effective.