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نسرين جهاد صدرالدين سدر

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​​​​​​​Transcriptome Analysis of Pseudomonas aeruginosa Biofilm Following the Exposure to Malaysian Stingless Bee Honey

Authors : ​Nesrin Seder, Mohd Hilmi Abu Bakar, and Walid Salem Abu Rayyan

Abstract : Introduction: Malaysian stingless bee honey (Trigona) has been aroused as a potential antimicrobial compound with antibiofilm activity. The capability of the gram-negative Bacillus P. aeruginosa to sustain a fatal infection is encoded in the bacterium genome. Methods: In the current study, a transcriptome investigation was performed to explore the mechanism underlying the biofilm dispersal of P. aeruginosa after the exposure to Trigona honey. Results: Microarray analysis of the Pseudomonas biofilm treated by 20% Trigona honey has revealed a down-regulation of 3478 genes among the 6085 screened genes. Specifically, around 13.5% of the down-regulated genes were biofilm-associated genes. The mapping of the biofilm-associated pathways has shown an ultimate decrease in the expression levels of the D-GMP signaling pathway and diguanylate cyclases (DGCs) genes responsible for c-diGMP formation. Conclusion: We predominantly report the lowering of c-di-GMP through the down regulation of DGC genes as the main mechanism of biofilm inhibition by Trigona honey.

Keywords : Pseudomonas, biofilm, stingless bee, Trigona, microarray, D-GMP

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​​​​​Phytochemical Investigation, Comparison and Characterization Study of Malaysian Stingless Bee Honey versus Jordanian Honey by LC-MS/MS

Authors : ​Nesrin Seder, Walid A.Rayyan, Wael A.Dayyih, Mohammad A. Al-Natour, and Abu Bakar Mohd Hilmi

Abstract : Phytochemical studies have revealed more than 200 distinct polyphenolic compounds present in honey alone. Stingless bee honey (Trigona honey) is naturally found in tropical and subtropical regions. This study aims to interrogate the physical parameters, polyphenolic content, and antioxidant characteristics of Malaysian Trigona honey in comparison with Apis honey (Centaurea hyalolepis)and Citrus honey from Jordan. The three honey types were subjected to phytochemical and chromatographic analysis to explore the differences in honey composition related to bees type and geographical location. The total phenolic content ranged between 288.09 and 663.19 mg GAE/kg of honey in the three honey types. Trigona honey had the highest phenolic content (663.19 mgGAE/kg) followed by Centaurea hyalolepis honey (471.87 mgGAE/kg), both of which were higher than Citrus honey (288.09 mgGAE/kg). Trigona honey showed an IC50 of 61.042 ± 0.45 mg/mL, whereas, Centaurea hyalolepis honey and Citrus honey had IC50 of 120.29 ± 1.64 mg/mL and 129.51 ± 4.3 mg/mL, respectively. Statistical analysis has revealed a significant negative correlation between the IC50 value for the three honey samples and the concentration of polyphenols (p≤0.001). Chromatographic analysis using LC-MS/MS showed a 28 and 42-fold difference in the polyphenolic content in Trigona over C. hyalolepis and Citrus honey, respectively. In conclusion, the diversity in the polyphenols contents and the high amounts of phytochemical compounds found in Trigona honey confers the antioxidant activity and there is no unique compound responsible for such activity over C. hyalolepis and Citrus honey.

Keywords : Trigona honey, Phytochemical, LC-MS/MS, Polyphenols, Jordan.

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​​​​​Wound healing, antimicrobial and anti-oxidant activity for Jordanian Juglans Regia L. unripe fruits

Authors : ​Afaf H. Al-Nadaf, Nesrin J. Seder, and Walid Abu Rayyan

Abstract : The present study, attempts to evaluate extraction efficiency of unripe fruit of J. regia in Jordan using different solvents with soxhlet and microwave extraction methods. We used five different types of solvents according to their increase in polarity to cover a wide range of phytochemical products. We examined the presence of some secondary metabolites as Tannins, Flavonoids, Saponins, and Alkaloids, as well as total phenols. Our results showed a higher efficiency for the microwave extraction method over the soxhlet extraction method in terms of higher yield amount and less time and solvents consuming. Water usage as an extraction solvent has generated the highest yield amount of 1.3 g in the defatted J. regia using microwave methods, whereas, the highest concentration of total phenols was demonstrated in the ethyl acetate extract of J. regia. In the antimicrobial susceptibility test, the ethyl acetate extract of J. regia showed a significant MIC inhibition concentration of 0.85 mg/ml against S. aureus, P. aeruginosa, and S. epidermidis. The healing activity of the ethyl acetate extract was as good as the reference drug (Baneocin) and even the ethyl acetate had an increase of 5% (P˂ 0.05) than reference drug.

Keywords : Juglans regia L, Unripe fruit, Wound healing, Microwave extraction, Antioxidant.

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​​​​​The Phytochemical Analysis and Antimicrobial Activity of Pergularia Tomentosa in North East Kingdom of Saudi Arabia KSA

Authors : Walid Abu Rayyan, Sami A. G. Alshammari, Abdulaziz M. F. AL-Sammary, Mutab S. S. ALShammari, Nisreen Seder, Luay F. Abu-Qatouseh, Muna Bostami, Kenza Mansoor, Mohammed F. Hamad, Ibrahim AlMajali and Wael Abu Dayyih​

Abstract : The increment in numbers of antimicrobial resistant strains along with the scarcity of new targets for drug industry has forced scientists to investigate deeply in the natural resources for new compounds with antimicrobial activity. Pergularia tomentosa is a member of the Apocynaceae family found in a wide geographical region including the Gulf region, Africa, and the Middle East. It is used as a remedy for the treatment of skin sores, asthma, and bronchitis. Dried plants of Pergularia tomentosa were subjected to extraction by using a Soxhlet extractor process to obtain essential oil and characterized by HPLC- Mass Spectroscopy (GC-MS). The essential oil was evaluated for antibacterial activity against pathogenic microorganisms by well diffusion method and confirmed by microdilution method. Additionally, we measured the antioxidant activity of the extracts using DPPH reagent. Phytochemical analysis has revealed variation in compositions and concentrations of P. tomentosa constituents grown in Hail from other agricultural regions. The lowest MIC was recorded with ethyl acetate extract MIC of 6.25 mg/ml against S. typhi, whereas, the ethanolic extract had the broadest effective against the five strains with a MIC of 25 mg/ml. In conclusion, we summarize a variation between the phytochemical constituents of P. tomentosa plants grown in the district of Hail and other geographical regions. In addition, there are several natural phytocompounds with an antimicrobial activity could be a good target for the antimicrobial and antioxidants industry.

Keywords : Antimicrobial; Antioxidants; HPLC-MS; Pergularia Tomentosa; Phytochemical.

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The Role of Glutamine-Rich Region of Candida Albicans Tec1p in Mediating Morphological Transition and Invasive Growth

Authors : W. Abu Rayyan, A. Singh, A. M. Al-Jaafreh, W. Abu Dayyih, M. Bustami, S. Salem, N. Seder, K. Schröppel​

Abstract : Hyphal growth and the transcriptional regulation to the host environment are key issues during the pathogenesis of C. albicans. Tec1p is the C. albicans homolog of a TEA transcription factor family, which share a conserved DNA-binding TEA domain in their N-terminal. In order to define a structure-function relationship of the C. albicans Tec1p protein, we constructed several mutations on the N terminal, C terminal or in the TEA binding domain itself by homologous recombination technology. The modifications in the open reading frame of TEC1 were tested for reconstitution of the morphogenetic development of the tec1/tec1 mutant strain CaAS12. Mutation in the TEA consensus sequence did not confer transition to hyphae whereas the reconstitution of the full-length Tec1p has reconstituted hyphal development. A deletion in one of glutaminerich regions either in the Tec1p N-terminal or the C-terminal in regions of 53-212 or 637–744 aa, respectively, did not restore morphological development in mutant CaAS12 strain. Whereas, the reconstitution with Tec1p mutants other than the glutamate-rich region has restored the morphogenetic switch. Additionally, the deletion of the glutamine-rich region has attenuated the invasive growth and the heat shock resistance of C. albicans. In conclusion, we show that a glutamine-rich region of Tec1p is essential for the hyphal development and mediating adaptation to the host environment of C. albicans.

Keywords : Candida albicans, transcription factor, TEA domain, hyphal formation, morphogenetic development, TEC1, Tet-induced.

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​​​​​Analytical and Comparative Study About the Impact of Lead Homeostasis on Cardiovascular Disorders in Humans

Authors : Eyad Mallah, Walid Abu Rayyan, Wael Abu Dayyih, Ibrahim S. Al-Majali, Haitham Qaralleh, Osama Yosef Al- Thunibat, Nisreen Seder, Mona Bustami, Luay Abu Qatoosah and Tawfiq Arafat​

Abstract : World health organization reported cardiovascular disease as the first cause of mortality for five consecutive years of 2010-2014. Smoking is an exogenous source of metals contamination in the human body and attributed to 12% of all deaths among adults aged over 30 years. This is the first study carried out to elucidate the contribution of the personal habits as smoking and the physiological process as aging on lead homeostasis and deposition in different storage sites of the human body especially on the cardiovascular system in Jordanian cadavers. 120 biological samples (coronary artery, ribs, and blood) were obtained from 40 Jordanian cadavers passed away in car accidents aged between 8 and 97. Samples were analyzed for lead levels using graphite atomic absorption. Mean lead concentrations for coronary artery (C-Pb), ribs (R-Pb) and blood (B-Pb) were 3.01 µg/g, 2.71 µg/g and 5.96 µg/dl, respectively. A significant correlation was demonstrated between both B-Pb and C-Pb levels (r= 0.273 P<0.05). A significant increment was demonstrated in lead levels in grade I and II of the atherosclerotic coronary arteries, whereas, there was no significance in the increase of lead levels in grades III, VI, and V. Duncan's test showed a significant correlation in levels of B-Pb and C-Pb with smoking and age. Eventually, lead intoxication emits an initiation effect on the endpoint cardiovascular disorders even though the blood lead levels were within recommended exposure range but still these levels are conveying deleterious side effects on the cardiovascular system.

Keywords : Atomic Absorption Cadavers; Coronary; Jordan; Ribs; Lead

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​​​​​Pseudomonas aeruginosa Virulence Factors and Antivirulence mechanisms to Combat Drug Resistance; A Systematic Review

Authors : Nesrin Seder, Walid Abu Rayyan, O'la Al-Fawares, Mohd Hilmi Abu Bakar​

Abstract :P. aeruginosaisan opportunistic microorganism that impose a versatile modifications in its genome to sustain an infection that can range from otitis media to a life threating disease. Moreover, antibiotic resistance endows P. aeruginosa characteristics to be one of the most resistant microorganisms to be treated. P. aeruginosa possess both cell associated and extracellular virulence factors. Cell associated virulence factors include flagellum, pili, adhesins, alginate, and lipopolysaccharide. While, extracellular virulence factors include hemolysins, proteases, lipases, exotoxin A, and cytotoxins

Keywords :P. aeruginosa, Virulence factors, Antimicrobial resistance, Biofilm

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​​​​​The Implications of COVID-19 Infection on Hematologic Parameters and Coagulation Activity: A Review

Authors : ​Walid Abu Rayyan, Waffaa Abu Hazzaa, Nesrin Seder, O'la Al-Fawares and Abdul Fattah Salah Fararjeh

Abstract :Wuhan, China, substantially is the epicenter of the COVID-19 pandemic in December 2019. Coronavirus, the confounder virus, a zoonotic in origin was the causative agent of the disseminated disease worldwide. Structural similarities and convergence points were demonstrated between the coronavirus, SARS, and MERS viruses. Aberrantly, a subset of patients developed a serious acute respiratory distress syndrome or diffuse alveolar injury whereas the rest of the patients encountered mild or no symptoms. The pathological clinical laboratory findings are not only critical in the diagnosis of the COVID-19 infection, on the contrary, but they are also crucial in the prognostic predictions about disease prognosis and therapeutic response. This review aims to give some historical context for the pandemic, demonstrate the laboratory's important role in the screening of COVID-19 infection, and review the current phase of biomarker examination in COVID-19 infection, focusing on markers derived directly from the hematological laboratory, reflecting the implications of COVID-19 on the hematological system and coagulation pathways. In conclusion, there is a direct significant correlation between infection severity, the death rate in COVID-19 patients, and the low number of either WBCs or a high number of WBCs with a low number of lymphocytes.

Keywords : COVID-19; Coagulation; D-Dimer; Hematology; Lymphopeni

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​​​​​A Comparative study for the Antimicrobial and Antibiofilm Activities of Trigona Honey against Opportunistic Microorganisms

Authors : ​Walid Aburayyan, Nesrin Seder and Abu Bakar Mohd Hilmi

Abstract :Biofilms formed by Pseudomonas aeruginosa and Streptococcus pyogenes are associated with devastating infections specifically in immunocompromised patients. The emergence of multidrug resistance in bacteria raised the ultimate need to establish an alternative drug to eradicate biofilm infections. Malaysian stingless bee honey (Trigona) has been aroused as a potential food with antibacterial and antibiofilm activities. However, there is limited knowledge about the effect of Trigona honey on the opportunistic bacteria during the early stage development of infections. Hence, this study aimed to evaluate the devastating effect of Trigona honey on P. aeruginosa and S. pyogenes growth and biofilm formation. The minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of Trigona honey were determined by the broth dilution method. Biofilm formation assay for P. aeruginosa and S. pyogenes was conducted using microtiter plates. Degradation assay for established biofilms was determined after the tested bacteria were exposed to 20% of Trigona honey. Trigona honey showed MIC and MBC values of 20% and 25%, respectively against both bacterial strains. Trigona honey successfully degraded 45.67% and 61.94% of P. aeruginosa and S. pyogenes biofilms, respectively. In conclusion Trigona honey can be used as a topical agent to prevent and treat bacterial infections.

Keywords : Trigona honey, Biofilm, Antibacterial, Malaysia.

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The C-terminal of Tec1p in Candida albicans Ameliorate Biofilm Formation and Generation time

Authors : Walid Abu Rayyan, Nesrin Seder, Klaus Schroppel​

Abstract :The morphogenetic switch from yeast to hyphae form under the regulation of transcription regulatory genes in response to the host environment is the major contributor to the pathogenesis of C. albicans. Tec1p of C. albicans, a transcription factor belonging to the TEA transcription factor family, harbors a conserved TEA domain for DNA binding in the N terminal of the transcription proteins. Tec1p is involved in mediating morphological change by activating different signaling pathways that lead to hyphal formation. The knockout strain tec1/tec1is not capable of hyphae development in vitro and is a virulent in vivo. To shed the light on the relationships between the structure and function of the C. albicans Tec1p protein, we examined several mutant TEC1 strains for biofilm formation and growth rate. The TEC1 recombination cassette was constructed by fusing the TEC1 ORFs behind a tetracycline promoter. The cassette is flanked with a homologous sequence of ADH1 promoter to facilitate homologous recombination into the promoter region of the ADH1 allele of the tec1/tec1 of the CaAs12 strain. The studies of biofilm formation and growth rate of TEC1mutants have revealed an essential role of CHAM (C-terminal Hyphal Activation Motif) in exerting virulence in Candida albicans. Herewith we conclude that the activity of Tec1p cannot be executed by autonomous activation of Tec1p itself but rather by a hetero dimeric complex formation with a cooperative partner which is not discovered until now. In conclusion, the C-terminal hyphal activation motif CHAM of Tec1p is essential for biofilm formation and virulence in vitro.

Keywords :TEC1, Candida albicans, Biofilm, Virulence genes, Fungi.

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The correlation of gene mutation of coagulopathy cascade with elevated D-dimer levels in COVID-19 patients

Authors :Mohammad Mansour Albalbaki, O’la AL-Fawares1, Walid Aburayyan, Nesrin Seder, Ola Mohammed Al-Sanabra, Lamya AL-Tahrawe, Mohammad Nazeeh Shatnawi ​

Abstract :SARS-CoV-2 had a pertinent implication on people’s daily lives and medical procedures during the pandemic or even after it. COVID-19 infections varied in the clinical aspect from mortality to asymptomatic episodes. The symptoms included fever, cough, difficulty breathing, loss of mobility, and chest pain. On the other hand, the patients who passed away from COVID-19 showed multiple organ failure, respiratory dysfunction, and disseminated coagulations throughout the body. D-dimer is a biomarker implemented in blood clotting and is commonly elevated in patients with thrombotic complications. This review sheds light on the correlation between gene mutations in the coagulopathy cascade among COVID-19 patients with high D-dimer levels. The elevated D-dimer levels are significantly associated with mutations in genes involved in the coagulopathy cascade. The findings suggest that these mutations may play an important role in developing thrombotic problems in COVID-19 patients. As a result, understanding the genetic basis of thrombotic development in COVID-19 could lead to new ways of avoiding and treating thrombotic problems

Keywords :COVID-19, hematology, coagulation, D-dimer, fibrinogen immunothrombosis

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Comparison of preservation enrichment media for long storage duration of Campylobacter jejuni

Authors : O'la AL-Fawares, Rozan O. Al-Khresieh, Walid Aburayyan, Nesrin Seder, Hala I Al-Daghistani, Nasser El-Banna, and Emran M. Abu-Taleb​

Abstract :Research on Campylobacter jejuni has been greatly impaired due to poor culturing and storage techniques. This study aims to evaluate different preservation methods for C. jejuni for long-term periods of preservation. Three different preservation methods (i, ii, and iii) were tested for their potential for long-term storage of C. jejuni which could be recommended as a simple and reliable method. In the first preservation method (i), 15% glycerol was added to 85% of the bacterial suspension of 4 McFarland standards (McF) and yeast extract. In the second method (ii) the bacterial suspension was supplemented with defibrinated lacked horse blood along with skim milk. Skim milk and 50% glycerol were added to the bacterial suspension in method (iii). Results indicated that method (ii) maintained the highest preservation rate of C. jejuni and the bacteria was capable to grow even after 12 months of storage at -80°C. Method (i) was less efficient than method (ii) where isolates eventually lost their ability to grow after 8 months. Method (iii) showed the least successful preservation rate as many isolates started to die after six months of preservation. This study emphasizes the importance of using defibrinated lacked horse blood along with skim milk during the preservation of bacterial isolates of C. jejuni.

Keywords :Campylobacter jejuni, defibrinated lacked horse blood, growth recovery, preservation methods, skim milk, ultra-cold mechanical freezers

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Probiotic therapy: A survey of Middle Eastern healthcare providers’ attitudes, beliefs, and practice patterns

Authors : ​O’la Ahmad AL-Fawares, Walid Abu Rayyan, Abdul Fattah Salah Fararjeh, Nesrin Seder

Abstract :This study compares Jordanian healthcare professionals to those in Syria, Lebanon, Palestine, Iraq, Egypt, the Gulf, and Morocco to assess their probiotic knowledge, attitudes, and practices. A cross-sectional study was carried out from May to August 2020 involving 269 healthcare providers (HCPs) from medical, dentistry, pharmacy, and nutrition majors. An online survey was used to collect data via email and social media. The analysis of variance test was used to examine if the variants reject the Null hypothesis. Multilingual linear test correlation coefficient was used to analyze the relationship between probiotic knowledge, attitude, and practice variables. Of all respondents, 13.3% of healthcare medical providers had good knowledge. The majority of HCPs that used probiotics were dietitians (69.4%). However, the multilingual linear test for probiotic knowledge, attitude, and practice correlated significantly with gender and specialty are 0.005 and 0.009, respectively. Most respondents had a limited level of knowledge of probiotics, despite the positive attitudes and positive practices toward probiotics, which could be improved by the implementation of targeted learning programs and workshops.

Keywords :Probiotics, Jordan, knowledge, attitude, practice, Middle East.

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​​​​​Research Title :Epidemiology, antibiotic resistance, and molecular detection of blaOXA and blaCTX-M Genes in ESBL-Producing Escherichia coli from urinary tract infections in Jordanian hospitals

Authors :Ayman Alsheikh, Abdalla Abuawwad, Israa Yousef, Dhia’a Abueswailem, Hafez Al-Momani, Walid Aburayyan, Nesrin Seder, Montasir Al-Mansi, Mohammad Al-Najjar, Lana Al-Shoubaki, Yaqeen Rjoub, Nawal Al-zaa’q ​

Abstract :Background: Urinary tract infections (UTIs) impose significant challenges among patients infected with multidrugresistant Escherichia coli (E. coli). The emergence of the extended-spectrum beta-lactamase (ESBL) strains complicates the treatment and exacerbates life-threatening infections. The comprehensive analysis of the resistance mechanisms combined with identifying the relevant genes amend the implications encountered with the devastating effects generated by the pathogenic microorganisms. This study sought about the prevalence of ESBL-producing E. coli in UTIs patients in Jordan, focusing on the prevalence of blaOXA and blaCTX-M resistance genes. Methods: A total of 500 urine samples were compiled from patients diagnosed with UTIs between November 2023 and May 2024. The samples were collected from five governmental hospitals distributed in three districts in Jordan (North, Middle, and South). Urine samples were cultured in the microbiology lab and the percentage of E. coli isolates was explored. Later on, isolates were tested for ESBL production using the double disk synergy test. Thus, molecular detection of the blaOXA and blaCTX-M genes was performed using polymerase chain reaction (PCR) assays. Statistical analysis was conducted to assess the prognostic correlation between the prevalence of resistance genes and variables such as location, age, and gender. Results In the current study, we investigated the prevalence of ESBL-producing E. coli in UTIs across Jordanian hospitals, focusing on the prevalence of the blaOXA and blaCTX-M genes with the correlation with the epidemiological factors. Our findings revealed an overall prevalence of 18% for ESBL- E. coli isolates, with the highest prevalence in the Northern governorates exhibiting 50% (OR=1.67, 95% CI: 1.103–2.519). Females showed a higher prevalence of the blaOXA gene (62.29%) compared to males (37.7%) (p=0.008). Moreover, the blaCTX-M gene was universally present in all samples with a prevalence of 98%. The multinomial regression analysis has revealed a significant correlation (p=0.001) between location on the distribution of the blaOXA gene among the study population. Conclusion The findings of the current study highlight alarming rates for the prevalence of ESBL-producing E. coli in UTIs across Jordan in general and in the northern districts as specific. Emphasizing the crucial need for effective surveillance, antibiotic stewardship, and infection control strategies. The significant regional variations in resistance rates suggest that localized factors, such as antibiotic misuse and environmental influences combined with the widespread of blaCTX-M and blaOXA genes point to the need for targeted antibiotic resistance surveillance in Jordan.

Keywords :Escherichia Coli (E. Coli), Urinary Tract Infection (UTI), Extended Spectrum β-Lactamase (ESBL), Jordan.

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​​​​​Research Title :Unveiling etiology and mortality risks in community-acquired pneumonia: A machine learning approach

Authors : Alaa Ali, Ahmad R Alsayed, Nesrin Seder, Yazun Jarrar, Raed H Altabanjeh, Mamoon Zihlif, Osama Abu Ata, Anas Samara, Malek Zihlif ​

Abstract :Community-acquired pneumonia (CAP) is associated with high mortality, and accurate diagnosis and risk prediction are essential for improving patient outcomes. Traditional diagnostic methods have limitations, prompting the use of machine learning (ML) to enhance diagnostic precision and treatment strategies. This study aims to develop ML models to predict CAP etiology and mortality using clinical data to enable early intervention. A retrospective cohort study was conducted on 251 adult CAP patients admitted to two Jordanian hospitals between March 2021 and February 2024. Various clinical data were analyzed using ML techniques, including linear regression, random forest, Shapley additive explanations (SHAP), lasso regression, mutual information analysis, logistic regression, and correlation analysis. Key predictors of CAP survival included zinc, vitamin C, enoxaparin, and insulin bolus. Mutual information analysis identified neutrophils, alanine transaminase, mean corpuscular volume, hemoglobin, and platelets as significant mortality predictors, while lasso regression highlighted meropenem, arterial blood gases, PCO2, and platelet count. Logistic regression confirmed intensive care unit (ICU) stay, pH, pulmonary severity index, white blood cell (WBC) count, and bicarbonate levels as crucial variables. Interestingly, lymphocyte count emerged as the strongest predictor of bacterial CAP, conflicting with established knowledge that associates neutrophils with bacterial infections. However, findings related to HCO3, blood urea nitrogen, and WBC levels were consistent with clinical expectations. SHAP analysis highlighted basophils and fever as key predictors. Further investigation is needed to resolve conflicting findings and optimize predictive models. ML offers promising applications for CAP prognosis but requires refinement to address discrepancies and improve reliability in clinical decision-making.

Keywords :Community-acquired pneumonia, CAP, machine learning, ML, mortality prediction, risk assessment, clinical predictors, SHAP analysis, logistic regression

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​​​​​Research Title :Improving HbA1c Levels by Methylcobalamin Vitamin in Diabetic Volunteers, Combined with Dapagliflozin as Type 2 Diabetes Mellitus Routine Treatment: A Controlled Randomized , Double-blind Trial

Authors : Walid Aburayyan, Zainab Zakaraya, Mohammed Hamad, Ibrahim S Majali, Wael Abu Dayyih, Nesrin Seder, Haneen Alkhadeir, Anas Khaleel ​

Abstract :Background: Diabetes mellitus is predominantly a growing global problem interconnected proportionally with obesity escalation. The current study evaluated the prognostic implications of vitamin B12 administration on Body Mass Index (BMI) and glycosylated hemoglobin (HbA1c) levels in type 2 diabetic patients treated with dapagliflozin. Methods: In this controlled randomized, double-blind trial, 160 patients for each arm were enrolled from July 2022 to June 2023 in Amman, Jordan.; 76 females and 84 males with inclusion criteria of vitamin B12 less than 233 ng/ml, age between 19-76 years, HbA1c range between 6.8-9.1%, and BMI less than 35. Group I received only dapagliflozin 10 mg/daily for a period of 12 months, whereas, group II received vitamin B12 supplements, methylcobalamin 500 µg, once daily with dapagliflozin 10 mg/day. HbA1c, Vitamin B12, and BMI were measured at time intervals of 0, 6, and 12 months. Using SPSS version 23, P values<0.05 were considered statistically significant. The continuous variables were reported as median and IQR. Mann-Whitney-u test and Correlations Spearman's rho were used for continuous variables. Results: The co-administration of vitamin B12 significantly decreased the levels of HbA1c in group II (54 participants) to 6.66±0.643 by 0.6 %, F(2,78)=172, P<0.001, compared to the subjects in group I (6.92±0.434). A significant impact of vitamin B12 administration on BMI lowering was observed at different time intervals during the study (P=0.002). Conclusion: The co-administration of vitamin B12 as a supplement for diabetic patients improved BMI and HbA1c levels.

Keywords :Body mass index; Dapagliflozin; Diabetes mellitus; Glycated hemoglobin; Vitamin B12

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​​​​​Research Title :Characterization of Antibiofilm and Antimicrobial Effects of Trigona Stingless Bee Honey Compared to Stinging Bee Centaurea hyalolepis and Citrus Honeys

Authors :Walid Salem Aburayyan, Nesrin Seder, O’la Al-fawares, AbdulFattah Fararjeh, Ibrahim S Majali, Yousef Al-Hajaya ​

Abstract :The antibiofilm and antimicrobial properties of tropical honey types including Malaysian stingless bee honey remain explicitly unexplored when compared with Apies honey. The antibiofilm and antimicrobial activities of the Malaysian Trigona honey were characterized with two stinging bee honey types (Centaurea hyalolepis and Citrus honeys) from Jordan. The antibiofilm and antimicrobial investigations were conducted on a set of seven microbial strains; five bacterial species of Pseudomonas aeruginosa ATCC 10145, Streptococcus pyogenes ATCC 19615, Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Klebsiella pneumoniae ATCC 13883, and two fungal strains Candida albicans ATCC 10231 and Candida krusei ATCC 14243. The antimicrobial investigations revealed a broad spectrum activity for Trigona honey against Gram-positive, Gram-negative, and fungal strains over the two honey types. One-way ANOVA showed a significant difference (p < 0.001) in the zone of inhibition ranging from 9 to 25 mm and minimum inhibition activity (MIC) ranged from 9.4–29.6% (w/v) against the microbial strains. Moreover, the addition of honey to established biofilms has induced a degradation activity in the biofilm mass. Two-way ANOVA showed a significant biofilm degradation proportion (p < 0.001) ranging from 1.3% to 91.3% following treatment with Trigona honey and the other honey types in relevance to the concentration ranging from 10% to 50% (w/v). Moreover, the antibiofilm activity was highly consistent with MIC affecting bacterial growth inhibition. In conclusion, a robust antimicrobial and antibiofilm activity for Trigona stingless bee honey over the stinging bee Centaurea hyalolepis and Citrus honeys is noticed which endows the usage of Trigona honey in the antimicrobial industry

Keywords : Trigona, antibiofilm, Centaurea hyalolepis, Citrus, honey

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​​​​​Research Title :The correlation of gene mutation of coagulopathy cascade with elevated D-dimer levels in COVID-19 patients

Authors :Mohammad Mansour Albalbaki, Walid Aburayyan, Nesrin Seder, Ola Mohammed Al-Sanabra, Lamya AL-Tahrawe, Mohammad Nazeeh Shatnawi ​

Abstract :SARS-CoV-2 had a pertinent implication on people’s daily lives and medical procedures during the pandemic or even after it. COVID-19 infections varied in the clinical aspect from mortality to asymptomatic episodes. The symptoms included fever, cough, difficulty breathing, loss of mobility, and chest pain. On the other hand, the patients who passed away from COVID-19 showed multiple organ failure, respiratory dysfunction, and disseminated coagulations throughout the body. D-dimer is a biomarker implemented in blood clotting and is commonly elevated in patients with thrombotic complications. This review sheds light on the correlation between gene mutations in the coagulopathy cascade among COVID-19 patients with high D-dimer levels. The elevated D-dimer levels are significantly associated with mutations in genes involved in the coagulopathy cascade. The findings suggest that these mutations may play an important role in developing thrombotic problems in COVID-19 patients. As a result, understanding the genetic basis of thrombotic development in COVID-19 could lead to new ways of avoiding and treating thrombotic problems.

Keywords :COVID-19, hematology, coagulation, D-dimer, fibrinogen immunothrombosis

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​​​​​Research Title :Probiotic therapy: a survey of Middle Eastern healthcare providers’ attitudes, beliefs, and practice patterns

Authors : Walid Abu Rayyan, Abdul Fattah Salah Fararjeh, Nesrin Seder ​

Abstract :This study compares Jordanian healthcare professionals to those in Syria, Lebanon, Palestine, Iraq, Egypt, the Gulf, and Morocco to assess their probiotic knowledge, attitudes, and practices. A cross-sectional study was carried out from May to August 2020 involving 269 healthcare providers (HCPs) from medical, dentistry, pharmacy, and nutrition majors. An online survey was used to collect data via email and social media. The analysis of variance test was used to examine if the variants reject the Null hypothesis. A multilingual linear test correlation coefficient was used to analyze the relationship between probiotic knowledge, attitude, and practice variables. Of all respondents, 13.3% of healthcare medical providers had good knowledge. The majority of HCPs that used probiotics were dietitians (69.4%). However, the multilingual linear test for probiotic knowledge, attitude, and practice correlated significantly with gender and speciality are 0.005 and 0.009, respectively. Most respondents had a limited level of knowledge of probiotics, despite the positive attitudes and positive practices toward probiotics, which could be improved by the implementation of targeted learning programs and workshops.

Keywords : Probiotics, Jordan, knowledge, attitude, practice, Middle East.

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​​​​​Research Title :Comparison of preservation enrichment media for long storage duration of Campylobacter jejuni

Authors : Rozan O Al-Khresieh, Walid Aburayyan, Nesrin Seder, Hala I Al-Daghistani, Nasser El-Banna, Emran M Abu-Taleb​

Abstract :Research on Campylobacter jejuni has been greatly impaired due to poor culturing and storage techniques. This study aims to evaluate different preservation methods for C. jejuni for long-term periods of preservation. Three different preservation methods (i, ii, and iii) were tested for their potential for long-term storage of C. jejuni which could be recommended as a simple and reliable method. In the first preservation method (i), 15% glycerol was added to 85% of the bacterial suspension of 4 McFarland standards (McF) and yeast extract. In the second method (ii) the bacterial suspension was supplemented with defibrinated lacked horse blood along with skim milk. Skim milk and 50% glycerol were added to the bacterial suspension in method (iii). Results indicated that method (ii) maintained the highest preservation rate of C. jejuni and the bacteria was capable to grow even after 12 months of storage at -80°C. Method (i) was less efficient than method (ii) where isolates eventually lost their ability to grow after 8 months. Method (iii) showed the least successful preservation rate as many isolates started to die after six months of preservation. This study emphasizes the importance of using defibrinated lacked horse blood along with skim milk during the preservation of bacterial isolates of C. jejuni.

Keywords : Campylobacter jejuni, defibrinated lacked horse blood, growth recovery, preservation methods, skim milk, ultra-cold mechanical freezers.

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​​​​​Research Title :A Comparative study for the Antimicrobial and Antibiofilm Activities of Trigona Honey against Opportunistic Microorganisms

Authors :Walid Aburayyan, Hilmi Abu Bakar, Nesrin Seder ​

Abstract :Biofilms formed by Pseudomonas aeruginosa and Streptococcus pyogenes are associated with devastating infections specifically in immunocompromised patients. The emergence of multidrug resistance in bacteria raised the ultimate need to establish an alternative drug to eradicate biofilm infections. Malaysian stingless bee honey (Trigona) has been aroused as a potential food with antibacterial and antibiofilm activities. However, there is limited knowledge about the effect of Trigona honey on the opportunistic bacteria during the early stage development of infections. Hence, this study aimed to evaluate the devastating effect ofTrigona honey on P. aeruginosa and S. pyogenesgrowth and biofilm formation. The minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of Trigona honey were determined by the broth dilution method. Biofilm formation assay for P. aeruginosa and S. pyogenes was conducted using microtiter plates. Degradation assay for established biofilms was determined after the tested bacteria were exposed to 20% of Trigona honey. Trigona honey showed MIC and MBC values of 20% and 25%, respectively against both bacterial strains. Trigona honey successfully degraded 45.67% and 61.94% of P. aeruginosa and S. pyogenes biofilms, respectively.In conclusion Trigona honey can be used as a topical agent to prevent and treat bacterial infections.

Keywords :Trigona honey, Biofilm, Antibacterial, Malaysia.

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​​​​​Research Title :The C-terminal of Tec1p in Candida albicans Ameliorate Biofilm Formation and Generation tim

Authors : Walid Abu Rayyan, Nesrin Seder, Klaus Schroppel ​

Abstract :The morphogenetic switch from yeast to hyphae form under the regulation of transcription regulatory genes in response to the host environment is the major contributor tothe pathogenesis of C. albicans. Tec1p of C. albicans, atranscription factor belonging to the TEA transcription factor family, harborsa conserved TEA domain for DNAbinding in the N terminal of the transcription proteins. Tec1p is involved in mediating morphological changeby activating different signalingpathways that lead to hyphal formation. The knockout strain tec1/tec1is not capable of hyphae developmentin vitro and is avirulent in vivo. To shed the light on therelationships between the structureand function of the C. albicans Tec1p protein, we examined several mutant TEC1 strains for biofilm formation and growth rate. The TEC1 recombination cassette was constructed by fusing the TEC1 ORFs behind a tetracycline promoter. The cassette is flanked with a homologous sequence of ADH1 promoter to facilitate homologous recombination into the promoter region of the ADH1 allele of the tec1/tec1 of the CaAs12 strain.The studies of biofilm formation and growth rate ofTEC1mutants have revealed an essential role of CHAM (C-terminal Hyphal Activation Motif) inexerting virulence in Candida albicans.Herewith we conclude that the activity of Tec1p cannot be executed by autonomous activation of Tec1p itself but rather by a heterodimeric complex formation with a cooperative partner which is not discovered until now. In conclusion, the C-terminal hyphal activation motif CHAM of Tec1p is essential for biofilm formation and virulence in vitro.

Keywords :TEC1, Candida albicans, Biofilm, Virulence genes, Fungi.

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​​​​​Research Title :The Implications of COVID-19 Infection on Hematologic Parameters and Coagulation Activity: A Review

Authors : Walid Abu Rayyan1, Waffaa Abu Hazzaa1, Nesrin Seder, O'la Al-Fawares and Abdul Fattah Salah Fararjeh​

Abstract :Wuhan, China, substantially is the epicenter of the COVID-19 pandemic in December 2019. Coronavirus, the confounder virus, a zoonotic in origin was the causative agent of the disseminated disease worldwide. Structural similarities and convergence points were demonstrated between the coronavirus, SARS, and MERS viruses. Aberrantly, a subset of patients developed a serious acute respiratory distress syndrome or diffuse alveolar injury whereas the rest of the patients encountered mild or no symptoms. The pathological clinical laboratory findings are not only critical in the diagnosis of the COVID-19 infection, on the contrary, but they are also crucial in the prognostic predictions about disease prognosis and therapeutic response. This review aims to give some historical context for the pandemic, demonstrate the laboratory's important role in the screening of COVID-19 infection, and review the current phase of biomarker examination in COVID-19 infection, focusing on markers derived directly from the hematological laboratory, reflecting the implications of COVID-19 on the hematological system and coagulation pathways. In conclusion, there is a direct significant correlation between infection severity, the death rate in COVID-19 patients, and the low number of either WBCs or a high number of WBCs with a low number of lymphocytes.

Keywords : COVID-19; Coagulation; D-Dimer; Hematology; Lymphopenia.

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​​​​​Research Title :Transcriptome Analysis of Pseudomonas aeruginosa Biofilm Following the Exposure to Malaysian Stingless Bee Honey

Authors : Nesrin Seder, Mohd Hilmi Abu Bakar, and Walid Salem Abu Rayyan​

Abstract :Introduction: Malaysian stingless bee honey (Trigona) has been aroused as a potential antimicrobial compound with antibiofilm activity. The capability of the gram-negative Bacillus P. aeruginosa to sustain a fatal infection is encoded in the bacterium genome. Methods: In the current study, a transcriptome investigation was performed to explore the mechanism underlying the biofilm dispersal of P. aeruginosa after the exposure to Trigona honey. Results: Microarray analysis of the Pseudomonas biofilm treated by 20% Trigona honey has revealed a down-regulation of 3478 genes among the 6085 screened genes. Specifically, around 13.5% of the down-regulated genes were biofilm-associated genes. The mapping of the biofilm-associated pathways has shown an ultimate decrease in the expression levels of the D-GMP signaling pathway and diguanylate cyclases (DGCs) genes responsible for c-diGMP formation. Conclusion: We predominantly report the lowering of c-di-GMP through the down regulation of DGC genes as the main mechanism of biofilm inhibition by Trigona honey.

Keywords :Pseudomonas, biofilm, stingless bee, Trigona, microarray, D-GMP

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​​​​​Research Title :Phytochemical Investigation, Comparison and Characterization Study of Malaysian Stingless Bee Honey versus Jordanian Honey by LC-MS/MS

Authors : ​Nesrin Seder, Walid A.Rayyan, Wael A.Dayyih, Mohammad A. Al-Natour, and Abu Bakar Mohd Hilmi

Abstract :Phytochemical studies have revealed more than 200 distinct polyphenolic compounds present in honey alone. Stingless bee honey (Trigona honey) is naturally found in tropical and subtropical regions. This study aims to interrogate the physical parameters, polyphenolic content, and antioxidant characteristics of Malaysian Trigona honey in comparison with Apis honey (Centaurea hyalolepis)and Citrus honey from Jordan. The three honey types were subjected to phytochemical and chromatographic analysis to explore the differences in honey composition related to bees type and geographical location. The total phenolic content ranged between 288.09 and 663.19 mg GAE/kg of honey in the three honey types. Trigona honey had the highest phenolic content (663.19 mgGAE/kg) followed by Centaurea hyalolepis honey (471.87 mgGAE/kg), both of which were higher than Citrus honey (288.09 mgGAE/kg). Trigona honey showed an IC50 of 61.042 ± 0.45 mg/mL, whereas, Centaurea hyalolepis honey and Citrus honey had IC50 of 120.29 ± 1.64 mg/mL and 129.51 ± 4.3 mg/mL, respectively. Statistical analysis has revealed a significant negative correlation between the IC50 value for the three honey samples and the concentration of polyphenols (p≤0.001). Chromatographic analysis using LC-MS/MS showed a 28 and 42-fold difference in the polyphenolic content in Trigona over C. hyalolepis and Citrus honey, respectively. In conclusion, the diversity in the polyphenols contents and the high amounts of phytochemical compounds found in Trigona honey confers the antioxidant activity and there is no unique compound responsible for such activity over C. hyalolepis and Citrus honey

Keywords : Trigona honey, Phytochemical, LC-MS/MS, Polyphenols, Jordan.

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​​​​​Research Title :Wound healing, antimicrobial and anti-oxidant activity for Jordanian Juglans Regia L. unripe fruits

Authors : ​Afaf H. Al-Nadaf, Nesrin J. Seder, and Walid Abu Rayyan

Abstract :The present study, attempts to evaluate extraction efficiency of unripe fruit of J. regia in Jordan using different solvents with soxhlet and microwave extraction methods. We used five different types of solvents according to their increase in polarity to cover a wide range of phytochemical products. We examined the presence of some secondary metabolites as Tannins, Flavonoids, Saponins, and Alkaloids, as well as total phenols. Our results showed a higher efficiency for the microwave extraction method over the soxhlet extraction method in terms of higher yield amount and less time and solvents consuming. Water usage as an extraction solvent has generated the highest yield amount of 1.3 g in the defatted J. regia using microwave methods, whereas, the highest concentration of total phenols was demonstrated in the ethyl acetate extract of J. regia. In the antimicrobial susceptibility test, the ethyl acetate extract of J. regia showed a significant MIC inhibition concentration of 0.85 mg/ml against S. aureus, P. aeruginosa, and S. epidermidis. The healing activity of the ethyl acetate extract was as good as the reference drug (Baneocin) and even the ethyl acetate had an increase of 5% (P˂ 0.05) than reference drug.

Keywords : Juglans regia L, Unripe fruit, Wound healing, Microwave extraction, Antioxidant.

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​​​​​Research Title :The Phytochemical Analysis and Antimicrobial Activity of Pergularia Tomentosa in North East Kingdom of Saudi Arabia KSA

Authors : Walid Abu Rayyan, Sami A. G. Alshammari, Abdulaziz M. F. AL-Sammary, Mutab S. S. ALShammari, Nisreen Seder, Luay F. Abu-Qatouseh, Muna Bostami, Kenza Mansoor, Mohammed F. Hamad, Ibrahim AlMajali and Wael Abu Dayyih​

Abstract :The increment in numbers of antimicrobial resistant strains along with the scarcity of new targets for drug industry has forced scientists to investigate deeply in the natural resources for new compounds with antimicrobial activity. Pergularia tomentosa is a member of the Apocynaceae family found in a wide geographical region including the Gulf region, Africa, and the Middle East. It is used as a remedy for the treatment of skin sores, asthma, and bronchitis. Dried plants of Pergularia tomentosa were subjected to extraction by using a Soxhlet extractor process to obtain essential oil and characterized by HPLC- Mass Spectroscopy (GC-MS). The essential oil was evaluated for antibacterial activity against pathogenic microorganisms by well diffusion method and confirmed by microdilution method. Additionally, we measured the antioxidant activity of the extracts using DPPH reagent. Phytochemical analysis has revealed variation in compositions and concentrations of P. tomentosa constituents grown in Hail from other agricultural regions. The lowest MIC was recorded with ethyl acetate extract MIC of 6.25 mg/ml against S. typhi, whereas, the ethanolic extract had the broadest effective against the five strains with a MIC of 25 mg/ml. In conclusion, we summarize a variation between the phytochemical constituents of P. tomentosa plants grown in the district of Hail and other geographical regions. In addition, there are several natural phytocompounds with an antimicrobial activity could be a good target for the antimicrobial and antioxidants industry.

Keywords : Antimicrobial; Antioxidants; HPLC-MS; Pergularia Tomentosa; Phytochemical

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​​​​​Research Title :The Role of Glutamine-Rich Region of Candida Albicans Tec1p in Mediating Morphological Transition and Invasive Growth

Authors : W. Abu Rayyan, A. Singh, A. M. Al-Jaafreh, W. Abu Dayyih, M. Bustami, S. Salem, N. Seder, K. Schröppel​

Abstract :Hyphal growth and the transcriptional regulation to the host environment are key issues during the pathogenesis of C. albicans. Tec1p is the C. albicans homolog of a TEA transcription factor family, which share a conserved DNA-binding TEA domain in their N-terminal. In order to define a structure-function relationship of the C. albicans Tec1p protein, we constructed several mutations on the N terminal, C terminal or in the TEA binding domain itself by homologous recombination technology. The modifications in the open reading frame of TEC1 were tested for reconstitution of the morphogenetic development of the tec1/tec1 mutant strain CaAS12. Mutation in the TEA consensus sequence did not confer transition to hyphae whereas the reconstitution of the full-length Tec1p has reconstituted hyphal development. A deletion in one of glutaminerich regions either in the Tec1p N-terminal or the C-terminal in regions of 53-212 or 637–744 aa, respectively, did not restore morphological development in mutant CaAS12 strain. Whereas, the reconstitution with Tec1p mutants other than the glutamate-rich region has restored the morphogenetic switch. Additionally, the deletion of the glutamine-rich region has attenuated the invasive growth and the heat shock resistance of C. albicans. In conclusion, we show that a glutamine-rich region of Tec1p is essential for the hyphal development and mediating adaptation to the host environment of C. albicans

Keywords : Candida albicans, transcription factor, TEA domain, hyphal formation, morphogenetic development, TEC1, Tet-induced

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​​​​​Research Title :Analytical and Comparative Study About the Impact of Lead Homeostasis on Cardiovascular Disorders in Humans

Authors :Eyad Mallah, Walid Abu Rayyan, Wael Abu Dayyih, Ibrahim S. Al-Majali, Haitham Qaralleh, Osama Yosef Al- Thunibat, Nisreen Seder, Mona Bustami, Luay Abu Qatoosah and Tawfiq Arafat ​

Abstract :World health organization reported cardiovascular disease as the first cause of mortality for five consecutive years of 2010-2014. Smoking is an exogenous source of metals contamination in the human body and attributed to 12% of all deaths among adults aged over 30 years. This is the first study carried out to elucidate the contribution of the personal habits as smoking and the physiological process as aging on lead homeostasis and deposition in different storage sites of the human body especially on the cardiovascular system in Jordanian cadavers. 120 biological samples (coronary artery, ribs, and blood) were obtained from 40 Jordanian cadavers passed away in car accidents aged between 8 and 97. Samples were analyzed for lead levels using graphite atomic absorption. Mean lead concentrations for coronary artery (C-Pb), ribs (R-Pb) and blood (B-Pb) were 3.01 µg/g, 2.71 µg/g and 5.96 µg/dl, respectively. A significant correlation was demonstrated between both B-Pb and C-Pb levels (r= 0.273 P<0.05). A significant increment was demonstrated in lead levels in grade I and II of the atherosclerotic coronary arteries, whereas, there was no significance in the increase of lead levels in grades III, VI, and V. Duncan's test showed a significant correlation in levels of B-Pb and C-Pb with smoking and age. Eventually, lead intoxication emits an initiation effect on the endpoint cardiovascular disorders even though the blood lead levels were within recommended exposure range but still these levels are conveying deleterious side effects on the cardiovascular system.

Keywords : Atomic Absorption Cadavers; Coronary; Jordan; Ribs; Lead

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