Anti-Inflammatory and Antioxidant Potential of Green Synthesized Iron Zinc Oxide (Fe0.25-ZnO) Nanoparticles of the Elaeagnus angustifolia.

Research interest in examining Elaeagnus angustifolia's potential as a source of anti-inflammatory and antioxidant agents has grown as a result of the plant's endorsement as a rich source of bioactive chemicals with promising anti-inflammatory and antioxidant activity.  In this study, zinc oxide (Fe0.25-ZnO) bimetallic nanoparticles (E.ang-Fe0.25-ZnO NPs) were synthesized using an aqueous extract of Elaeagnus angustifolia. Synthesized Fe0.25-ZnO nanoparticles were characterized by FTIR and XRD. The anti-inflammatory and antioxidant activities were investigated in LPS-stimulated RAW 264.7 macrophages using RT-PCR and ELISA techniques for antioxidant- and inflammation-related genes. The concentration of 39.6µg/ml of E.ang-Fe0.25-ZnO NPs demonstrated a significant anti-inflammatory activity by suppressing the mRNA levels of TNF-α and IL-6 by 88.3%±1.9 and 93.6%±0.1, respectively, compared to LPS-stimulated cells.  This was confirmed by the significant reduction of TNF-α and IL-6 secretion levels from 95.2 and 495.6 pg/ml in LPS-stimulated cells to 5.6 and 26.5 pg/ml in E.ang-Fe0.25-ZnO treated group. In addition, E.ang-Fe0.25-ZnO NPs nanoparticles treatment significantly enhanced the expression of antioxidant-related genes, SOD and CAT. Together, our results proved that phyto-mediated Fe0.25-ZnO nanoparticles using Elaeagnus angustifolia have great potential in biomedical applications such as anti-inflammatory and antioxidant.

Enhancing chemosensitivity of PANC1 pancreatic cancer cells to gemcitabine using ANGTPL4, Notch1 and NF-κß1 siRNAs.

Aim: siRNA can silence targeted genes with lesser toxicity than therapeutic drugs. Therefore, this study aims to investigate new approaches to treat pancreatic cancer (PC) using combinations of siRNA and gemcitabine. Methods: Three genes, ANGTPL4, Notch1 and NF-κß1, were silenced using siRNA, and their anti-proliferative effects were studied in combination with gemcitabine on pancreatic cancer cell line (PANC-1) using MTT viability assay. Results: Our results showed a significant reduction in PANC-1 cells growth upon treating cells with gemcitabine and single and combinations of siRNA sequences specific for ANGTPL4, Notch1 and NF-κß1 genes. Conclusion: Co-transfection of gemcitabine-treated PANC-1 cells with ANGPTL4, Notch1 and NF-κßsiRNAs enhances the chemosensitivity of PANC-1 cells to gemcitabine can be a promising therapeutic approach.

Pancreatic cancer (PC) is prominent with its aggressive behavior and metastatic properties, making it one of the leading causes of cancer-related deaths worldwide. PC is associated with poor prognosis and low survival rate, with 5 years survival rate of less than 9%. Moreover, only 20% of PC patients could undergo surgery, which makes investigating new therapeutic approaches to treat PC necessary. In the current study, the chemosensitivity of pancreatic cancer cells to gemcitabine has been enhanced using a single and combination of ANGTPL4, Notch1 and NF-κß1 siRNA.

The Association of M235T Genetic Polymorphism in Angiotensinogen Gene and Other Non-Genetic Factors with Essential Hypertension among Jordanian Patients.

BACKGROUND: Hypertension, characterized by elevated pressure, poses a significant health risk. Recent studies in Jordan highlight high hypertension rates, emphasizing the need for genetic investigations to comprehend essential hypertension determinants. The AGT gene, part of the Renin Angiotensin System, is linked to blood pressure regulation. Limited information exists on the frequency of this polymorphism among Jordanian hypertensive patients. AIMS: This study explores the association between the AGT M235T polymorphism and essential hypertension in Jordan. METHODS: A cross-sectional study with 435 participants (199 hypertensive, 236 non-hypertensive) was conducted at the University of Jordan Hospital. Blood pressure was measured, and genetic analysis of the AGT M235T polymorphism was completed using the PCR-RFLP technique. Chi-square and t-tests were used for comparisons using SPSS software. RESULTS: Hypertensive patients exhibited significantly higher weight, BMI, and blood pressure. Genotyping results showed no significant difference (p > 0.05, Chi-square) in AGT M235T polymorphism distribution between control and patient groups. In addition, allele frequencies showed comparable patterns (p > 0.05, Chi-square). All genotype frequencies showed no deviation from the Hardy-Weinberg equation (p > 0.05, Chi-square). CONCLUSIONS: The AGT M235T genetic polymorphism is not more prevalent among hypertensive patients in Jordan, although the average weight and BMI among hypertensive patients is higher than the non-hypertensive participants. Obesity can be addressed as a potential risk factor for essential hypertension in Jordan. In addition, it is recommended to find out the influence of the AGT M235T genetic polymorphism on the response of antihypertensive drugs among hypertensive patients in Jordan.

Anti­angiogenic and cytotoxic evaluation of green­synthesized Fe2ZnO4 nanoparticles against MCF­7 cell line.

The use of plants for nanoparticle (NP) synthesis, grounded in green chemistry principles, is an environmentally friendly and economically viable approach. In the present study, the leaf extract of Elaeagnus angustifolia L. was used as a biosynthetic agent to generate bimetallic zinc oxide NPs. The present study investigated the effect of ZnO NPs on anti-angiogenesis and cell migration. Various bimetallic NPs, including zinc-iron oxide and nickel-zinc oxide, underwent characterization through Fourier-transform infrared spectroscopy and X-ray Diffraction within the 25-65˚ range. Confirmation of NP formation was determined by identifying the surface plasmon resonance peak. MTT assay was used to determine the cytotoxic properties of E. angustifolia L. extracts, ZnO NPs and associated metals in MCF-7 breast cancer cells. The plant extract demonstrated antiproliferative effects at 200 µg/ml, whereas E. ang-Fe2ZnO4 NPs showed varying cytotoxic effects based on concentration. The rat aortic ring and cell migration assays illuminated anti-angiogenic attributes, with the E. ang-Fe2ZnO4 NPs blocking blood vessel development entirely at 100 µg/ml, implying profound anti-angiogenic efficacy. Therefore, E. ang-Fe2ZnO4 NPs may serve a role in antiangiogenic therapy.

Effect of quercetin on doxorubicin cytotoxicity in sensitive and resistant human MCF7 breast cancer cell lines.

Chemoresistance is the major cause of cancer recurrence, relapse and eventual death. Doxorubicin resistance is one such challenge in breast cancer. The use of quercetin, an antioxidant, in combination with doxorubicin has been investigated for offering protection to normal cells from the toxic side effects of doxorubicin in addition to modulation of its resistance. The present study aimed to investigate the effects of quercetin in prevention of a doxorubicin-chemoresistant phenotype in both doxorubicin-sensitive and -resistant human MCF-7 breast cancer cell lines. A doxorubicin-resistant MCF-7 cell line was established. The development of resistant cells was closely monitored for changes in morphological features. Sensitivity to doxorubicin and the doxorubicin/quercetin combination was assessed using the tetrazolium assay. To determine the mechanism by which quercetin sensitizes the doxorubicin MCF-7-resistant cell line to doxorubicin, gene expression alterations in breast cancer-related genes were examined using the reverse transcription-quantitative PCR (RT-qPCR) array technology. Resistant MCF cells were successfully developed and the inhibitory concentration (IC50) value of doxorubicin increased from 0.133 to 4 µM (wild-type to resistant). The effects of the quercetin/doxorubicin combination exhibited different effects on wild-type vs. resistant cells. The IC50 of doxorubicin was reduced in wild cells, whereas resistant cells showed an increase in cell viability at lower concentrations and a potentiation of the effects of doxorubicin only at higher concentrations. Annexin V/propidium iodide staining demonstrated that quercetin drives cells into late apoptosis and necrosis, but in resistant cells, necrosis predominates. RT-qPCR results revealed that quercetin led to a reversal in doxorubicin effects via up- and downregulation of important genes such as SNAI2, PLAU and CSF1 genes. Downregulation of cell migration genes, SNAI2 (-31.23-fold) and plasminogen activator, urokinase (PLAU; -30.62-fold), and the apoptotic pathway gene, colony stimulating factor 1 (CSF1; -17.25-fold) were the most important querticin-associated events. Other gene alterations were also observed involving cell cycle arrest and DNA repair pathways. The results of the present study indicated that quercetin could lead to a reversal of doxorubicin resistance in breast cancer cells via downregulation of the expression of important genes, such as SNAI2, PLAU and CSF1. Such findings may represent a potential strategy for reversing breast cancer cell-related chemoresistance.

Anti-Inflammatory and Antioxidant Effects of Rosmarinic Acid Trimetallic (Cu0.5Zn0.5Fe2O4) Nanoparticles.

Newly, green metallic-nanoparticles (NPs) have received scientists' interest due to their wide variable medicinal applications owned to their economical synthesis and biologically compatible nature. In this study, we used rosmarinic acid (RosA) to prepare Cu0.5Zn0.5FeO4 NPs and later encapsulated them using PEG polymer. Characterization of NPs was done using the XRD method and SEM imaging. Further, we explored the encapsulated NPs for anti-inflammatory properties by downregulating the expression of pro-inflammatory cytokines mRNA in LPS-stimulated Raw 264.7 cells. Besides, employing DPPH, NO and ABTS radical scavenging assays to examine the antioxidant activity of the synthesized Cu0.5Zn0.5FeO4 NPs. Cu0.5Zn0.5FeO4 NPs revealed moderate antioxidant activity by scavenging DPPH and nitric oxide. We demonstrated that the NPs showed high potential anti-inflammatory activity by suppressing the mRNA and protein levels of pro-inflammatory cytokines in a dose-dependent manner, in LPS-induced Raw 264.7 cells. To our best knowledge, this is the first report where RosA was found to be a suitable phyto source for the green synthesis of Cu0.5Zn0.5FeO4 NPs and their in vitro anti-inflammatory and antioxidant effects. Taken together, our findings suggest that the RosA is a green resource for the eco-friendly synthesis of Cu0.5Zn0.5FeO4/PEG NPs, which further can be employed as a novel anti-inflammatory therapeutic agent.

Alteration in the expression of the chemotherapy resistance­related genes in response to chronic and acute hypoxia in pancreatic cancer.

Pancreatic cancer is currently one of the least curable types of human cancer and remains a key health problem. One of the most important characteristics of pancreatic cancer is its ability to grow under hypoxic conditions. Hypoxia is associated with resistance of cancer cells to radiotherapy and chemotherapy. It is a major contributor to pancreatic cancer genetic instability, which local and systemic resistance that may result in poor clinical outcome. Accordingly, identifying gene expression changes in cancer resistance genes that occur under hypoxic conditions may identify a new therapeutic target. The aim of the present study was to explore the association between hypoxia and resistance to chemotherapy and determine the alteration in the expression of cancer resistance-related genes in the presence of hypoxia. Pancreatic cancer cells (PANC-1) were exposed to 8 h hypoxic episodes (<1% oxygen) three times/week for a total of 20 episodes (chronic hypoxia) or 72 h hypoxic episodes twice/week for a total of 10 episodes (acute hypoxia). The alterations in gene expression were examined using reverse transcription-quantitative PCR array compared with normoxic cells. Chemoresistance of hypoxic cells toward doxorubicin was assessed using MTT cell proliferation assay. Both chronic and acute hypoxia induced chemoresistance toward doxorubicin in PANC-1 pancreatic cancer cell line. The greatest changes occurred in estrogen Receptor Alpha Gene (ESR1) and ETS Like-1 protein (ELK1) pathways, in nucleic transcription factor Peroxisome proliferator-activated receptors (PPARs) and in a cell cycle inhibitor cyclin dependent kinase inhibitor 1A (CDKN1A). The present study demonstrated that exposing cells to prolonged hypoxia results in different gene expression changes involving pleotropic pathways that serve a role in inducing resistance in pancreatic cancer.

The first study characterizing the respiratory microbiome in cystic fibrosis patients in Jordan

Cystic fibrosis (CF) is most commonly seen in Caucasians and is uncommon in the Middle East. This study, based in Jordan, aimed to describe the association between lung exacerbation in CF patients and the respiratory microbiome composition. Using the 16S rRNA marker-gene sequencing, we investigated the microbiota in sputa during exacerbation (E1) and 14 days after the exacerbation (E2) of two CF patients admitted to the hospital. Detected genera with high abundance in the E1-related sputa of the first patient included Achromobacter and Streptococcus. At E2, Achromobacter and Staphylococcus were the highest abundant genera. Regarding the second patient, Veillonella and Streptococcus, were the highest abundant genera at E1. Whereas, Streptococcus and Veillonella were the highest abundant genera. This is the first study, based in Jordan, to report and describe the respiratory microbiome during and after the exacerbation of CF patients. This study suggests that pulmonary exacerbation in CF patients can result in alterations in their respiratory microbiome. A better knowledge of this link could allow more focused use of antibiotics, especially during exacerbations, improving clinical efficacy and patient outcomes. (AU)

Differential miRNA expression of hypoxic MCF7 and PANC-1 cells.

Background: Hypoxia plays a critical role in the tumor microenvironment by affecting cellular proliferation, metabolism, apoptosis, DNA repair, and chemoresistance. Since hypoxia provokes a distinct shift of microRNA, it is important to illustrate the relative contribution of each hypoxamiR to cancer progression. Aims: The present study aims to shed light on the hypoxamiRs that are involved in pancreatic and breast cancer progression to highlight novel targets for the development of new therapies. Methods: For 20 cycles, MCF7 breast cancer cells and PANC-1 pancreatic cancer cells were subjected to chronic cyclic hypoxia, which consisted of 72 hours of hypoxia followed by 24 hours of reoxygenation. After 10 and 20 cycles of hypoxia, miRNA expression alterations were profiled using RT-PCR array and further analyzed using a visual analytics platform. The MTT cell proliferation assay was used to determine hypoxic cells' chemoresistance to doxorubicin. Results: Under chronic cyclic hypoxia, hypoxic PANC-1 cells have a comparable doubling time with their normoxic counterparts, whereas hypoxic MCF7 cells show a massive increase in doubling time when compared to their normoxic counterparts. Both hypoxic cell lines developed EMT-like phenotypes as well as doxorubicin resistance. According to the findings of miRNet, 6 and 10 miRNAs were shown to play an important role in enriching six hallmarks of pancreatic cancer in the 10th and 20th cycles of hypoxia, respectively, while 7 and 11 miRNAs were shown to play an important role in enriching the four hallmarks of breast cancer in the 10th and 20th cycles of hypoxia, respectively. Conclusions: miR-221, miR-21, miR-155, and miR-34 were found to be involved in the potentiation of hypoxic PANC-1 hallmarks at both the 10th and 20th cycles, while miR-93, miR-20a, miR-15, and miR-17 were found to be involved in the potentiation of hypoxic MCF7 hallmarks at both the 10th and 20th cycles. This variation in miRNA expression was also connected to the emergence of an EMT-like phenotype, alterations in proliferation rates, and doxorubicin resistance. The chemosensitivity results revealed that chronic cyclic hypoxia is critical in the formation of chemoresistant phenotypes in pancreatic and breast cancer cells. miR-181a and let-7e expression disparities in PANC1, as well as miR-93, miR-34, and miR-27 expression disparities in MCF7, may be associated with the formation of chemoresistant MCF7 and PANC-1 cells following 20 cycles of chronic cyclic hypoxia. Indeed, further research is needed since the particular mechanisms that govern these processes are unknown.

CRHR1 polymorphism at rs242941, rs242940, and rs72834580: association of symptoms improvement with intranasal corticosteroids in allergic rhinitis Jordanian patients.

OBJECTIVES: Rhinitis is classified into several types with allergic rhinitis (AR) being the most common. AR is among the inflammatory diseases, such as asthma and chronic obstructive pulmonary disease (COPD), in which corticosteroids are administered to overcome the decrease in cortisol production. The treatment options available for AR vary with 1st line treatment being intranasal corticosteroids (INCS). The responsiveness to corticosteroids is due to their binding to corticotropin-releasing hormone receptor-1 (CRHR1). Various studies have studied the responsiveness to corticosteroids treatment in patients with asthma and COPD in association with CRHR1 gene single nucleotide polymorphisms (SNPs). METHODS: In our study, we investigated the association of three SNPs of CRHR1 gene (rs242941, rs242940, and rs72834580) with symptoms improvement post-treatment in AR patients. Blood samples were collected from 103 patients for DNA extraction and gene sequencing. Those patients started to receive INCS for 8 weeks and their symptoms were assessed, through a questionnaire, before treatment and post-treatment to check for symptoms improvement. RESULTS: Our data showed that improvement of eye redness is significantly less following INCS treatment in patients with allele (C) (AOR=0.289, p-value-0.028, 95 % CI=0.096-0.873) and genotype (CC) (AOR=0.048, p-value-0.037, 95 % CI=0.003-0.832) of rs242941 SNP. There was no correlation with other genotypes, alleles, or haplotypes of the investigated SNPs. CONCLUSIONS: Our findings show that there is no correlation between CRHR1 gene polymorphism and symptoms improvement following INCS treatment. Further studies are required to evaluate the association of INCS and symptoms improvement post-treatment with larger sample size.

Alteration of the Respiratory Microbiome in Hospitalized Patients with Asthma-COPD Overlap during and after an Exacerbation.

The immediate aim of this study was to comparatively examine the bacterial respiratory microbiome of patients in a stable state and during an exacerbation of asthma-COPD (chronic obstructive pulmonary disease) overlap (ACO). This prospective observational study took place in Jordan between 1 September 2021 and 30 April 2022. Sputum samples from patients with recognized ACO were acquired within 48 h of the exacerbation onset and again at 3 weeks following the exacerbation. The next-generation sequencing Illumina MiSeq was employed and uncovered significantly high bacterial diversity in the sputa. The results showed a significant decrease in the taxonomic richness in the sputum samples collected during the exacerbation episodes compared with those collected from patients in a stable state (p = 0.008), with an increase in the taxonomic evenness (p < 0.005). This change in the composition of the airway bacterial community suggests that the replacement of a significant portion of the airway microbiome with certain microorganisms may play a role in the decrease in microbial diversity observed during an ACO exacerbation. Greater knowledge of this link could allow for a more focused administration of antibiotics, especially during exacerbations, improving clinical efficacy and patient outcomes.

Phytochemical Analysis and Anticancer Properties of Drimia maritima Bulb Extracts on Colorectal Cancer Cells.

Cancer is a worldwide health problem and is the second leading cause of death after heart disease. Due to the high cost and severe side effects associated with chemotherapy treatments, natural products with anticancer therapeutic potential may play a promising role in anticancer therapy. The purpose of this study was to investigate the cytotoxic and apoptotic characteristics of the aqueous Drimia maritima bulb extract on Caco-2 and COLO-205 colorectal cancer cells. In order to reach such a purpose, the chemical composition was examined using the GC-MS method, and the selective antiproliferative effect was determined in colon cancer cell lines in normal gingival fibroblasts. The intracellular ROS, mitochondrial membrane potential, and gene expression changes in selected genes (CASP8, TNF-α, and IL-6 genes) were assessed to determine the molecular mechanism of the antitumor effect of the extract. GC-MS results revealed the presence of fifty-seven compounds, and Proscillaridin A was the predominant secondary metabolite in the extract. The IC50 of D. maritima bulb extract on Caco-2, COLO-205, and the normal human gingival fibroblasts were obtained at 0.9 µg/mL, 2.3 µg/mL, and 13.1 µg/mL, respectively. The apoptotic effect assay indicated that the bulb extract induced apoptosis in both colon cancer cell lines. D. maritima bulb extract was only able to induce statistically significant ROS levels in COLO-205 cells in a dose-dependent manner. The mitochondrial membrane potential (MMP) revealed a significant decrease in the MMP of Caco-2 and COLO-205 to various concentrations of the bulb extract. At the molecular level, RT-qPCR was used to assess gene expression of CASP8, TNF-α, and IL-6 genes in Caco-2 and COLO-205 cancer cells. The results showed that the expression of pro-inflammatory genes TNF-α and IL-6 were upregulated. The apoptotic initiator gene CASP8 was also upregulated in the Caco-2 cell line and did not reach significance in COLO-205 cells. These results lead to the conclusion that D. maritima extract induced cell death in both cell lines and may have the potential to be used in CRC therapy in the future.

Molecular Accounting and Profiling of Human Respiratory Microbial Communities: Toward Precision Medicine by Targeting the Respiratory Microbiome for Disease Diagnosis and Treatment.

The wide diversity of microbiota at the genera and species levels across sites and individuals is related to various causes and the observed differences between individuals. Efforts are underway to further understand and characterize the human-associated microbiota and its microbiome. Using 16S rDNA as a genetic marker for bacterial identification improved the detection and profiling of qualitative and quantitative changes within a bacterial population. In this light, this review provides a comprehensive overview of the basic concepts and clinical applications of the respiratory microbiome, alongside an in-depth explanation of the molecular targets and the potential relationship between the respiratory microbiome and respiratory disease pathogenesis. The paucity of robust evidence supporting the correlation between the respiratory microbiome and disease pathogenesis is currently the main challenge for not considering the microbiome as a novel druggable target for therapeutic intervention. Therefore, further studies are needed, especially prospective studies, to identify other drivers of microbiome diversity and to better understand the changes in the lung microbiome along with the potential association with disease and medications. Thus, finding a therapeutic target and unfolding its clinical significance would be crucial.

Discovery of new potent lysine specific histone demythelase-1 inhibitors (LSD-1) using structure based and ligand based molecular modelling and machine learning.

Lysine-specific histone demethylase 1 (LSD-1) is an epigenetic enzyme that oxidatively cleaves methyl groups from monomethyl and dimethyl Lys4 of histone H3 and is highly overexpressed in different types of cancer. Therefore, it has been widely recognized as a promising therapeutic target for cancer therapy. Towards this end, we employed various Computer Aided Drug Design (CADD) approaches including pharmacophore modelling and machine learning. Pharmacophores generated by structure-based (SB) (either crystallographic-based or docking-based) and ligand-based (LB) (either supervised or unsupervised) modelling methods were allowed to compete within the context of genetic algorithm/machine learning and were assessed by Shapley additive explanation values (SHAP) to end up with three successful pharmacophores that were used to screen the National Cancer Institute (NCI) database. Seventy-five NCI hits were tested for their LSD-1 inhibitory properties against neuroblastoma SH-SY5Y cells, pancreatic carcinoma Panc-1 cells, glioblastoma U-87 MG cells and in vitro enzymatic assay, culminating in 3 nanomolar LSD-1 inhibitors of novel chemotypes.

GSDM gene polymorphisms regulate the IgE level in asthmatic patients.

BACKGROUND: Gasdermin A (GSDMA) and gasdermin B (GSDMB) have been associated with childhood and adult asthma in many populations including the Jordanian population. It is also known that IgE plays a crucial role in various allergic disorders, such elevated levels of total serum IgE were detected in asthma and allergic rhinitis. IgE immunoglobulin is responsible for the release of numerous inflammatory mediators, such as histamine and prostaglandins, from mast cells in asthmatic patients. OBJECTIVE: In this study, single nucleotide polymorphisms of GSDMA (rs7212938, T/G) and GSDMB (rs7216389, T/C) in Jordanian population were investigated for their association with total IgE levels in serum of asthmatic children and adult subjects. METHODS: The genetic polymorphism analysis for SNPs was performed using the polymerase chain reaction (PCR)/restriction fragment length polymorphism method (RFLP). Three analysis models were applied to the genotype data: co-dominant, dominant and recessive. RESULTS: Our data demonstrate a significant correlation between GSDMB genetic SNP (rs7216389) and the total IgE serum level. Where one minor allele in the GSDMB gene is sufficient to induce significant changes in the IgE serum levels and plays a role in the pathogenesis of asthma in asthmatic children of the Jordanian population. Suggesting that this polymorphism might have a protective effect against asthma risk. While the presence of the GSDMB polymorphism alone might not be sufficient to associate with the high risk of developing asthma or responding to it in adults in Jordanian population. CONCLUSION: In conclusion, the current study confirms the significant association of GSDMB genetic SNP (rs7216389) with IgE levels in asthma patients in Jordanian population, while no significant correlation of GSDMA and IgE level was found in both child and adult asthmatic patients.

Anti-Proliferative Effect of Potential LSD1/CoREST Inhibitors Based on Molecular Dynamics Model for Treatment of SH-SY5Y Neuroblastoma Cancer Cell Line.

BACKGROUND: Lysine-specific demethylase is a demethylase enzyme that can remove methyl groups from histones H3K4me1/2 and H3K9me1/2. It is expressed in many cancers, where it impedes differentiation and contributes to cancer cell proliferation, cell metastasis and invasiveness, and is associated with inferior prognosis. LSD1 is associated with its corepressor protein CoREST, and utilizes tetrahydrofolate as a cofactor to accept CH2 from the demethylation process. The fact that the cofactor is best bound to the active site inspired us to explore its interactions to LSD1/CoREST enzyme complex utilizing molecular dynamics simulation, which aids designing novel and potent inhibitors. OBJECTIVE: In this study we minted to identify a new potential LSD1/CoREST inhibitors and test the potency and the safety of such inhibitors against human neuroblastoma and fibroblast cells lines. METHODS: We have implemented a previously derived model from the molecular dynamics simulation study and the key contacts to the active site in a subsequent structure based drug design and in-silico screening, which revealed a number of potential inhibitors toward LSD1/CoREST complex. The anti-proliferative activities of the identified compounds will be tested against neuroblastoma SH-SY5Y cancer cell line which known to highly express LSD1/CoREST complex. RESULTS: In-silico mining on National Cancer Institute (NCI) database identified 55 promising and structurally diverse inhibitors. Applying the abovementioned molecular modeling procedure yielded four compounds of LSD1/CoREST inhibiters with IC50 < 2µM. The four lead compounds were tested against SH-SY5Y neuroblastoma cell line that known to express high level of LSD1 and illustrated a potent activity with an IC50 ranging from 0.195 to 1.52µM. To estimate the toxicity of the selective leads, they were tested against normal fibroblast cells and scored a relatively high IC50 ranging from 0.303 to ≥ 100µM. CONCLUSION: Our model revealed promising inhibitors that can be used in treating cancers that overexpress the LSD1 enzyme such as the SH-SY5Y neuroblastoma.

Effects of cyclic acute and chronic hypoxia on the expression levels of metabolism related genes in a pancreatic cancer cell line.

The aim of this study was to characterize cycling hypoxia-induced changes in the expression of metabolism-related genes in the pancreatic cancer cell line PANC1. PANC1 cells were exposed to either 7 h cycles of hypoxia every other day for 20 cycles (cyclic acute hypoxia), or for 72 h cycles of hypoxia once a week for 5 cycles (cyclic chronic hypoxia). Changes in gene expression were profiled using reverse transcription-quantitative PCR and compared to cells cultured under normoxic conditions. Western blotting analysis confirmed upregulation of HIF1-α, glucose-6-phosphate isomerase, and ribokinase at the mRNA level. Upregulation in genes encoding enzymes involved in glycolysis was greater in cells cultured under cyclic acute hypoxia compared with cells cultured under chronic hypoxia including hexokinase2 and phosphoglycerate kinase 1. Genes encoding the pentose phosphate pathway (PPP) enzymes (transketolase and transaldolase) were upregulated to a similar degree. The expression of genes encoding pyruvate dehydrogenases that block pyruvate flow to the TCA cycle was significantly upregulated. Thus, exposure of PANC1 cells to acute hypoxia resulted in the upregulation of genes that shift the metabolism of cells towards glycolysis and the pentose phosphate pathway (PPP) in adaptation to hypoxic stress.

Investigating the human rhinovirus co-infection in patients with asthma exacerbations and COVID-19.

Objective: To investigate the co-infections with human rhinovirus (HRV) among patients with asthma exacerbation and COVID-19 in Jordan. Also, to determine the frequency of acute asthma exacerbation before and during the COVID-19 pandemic on a matched basis. Methods: The data of this prospective cohort research consisted of clinical variables. During the first visit, and after 14-days, nasopharyngeal swabs were taken and the quantitative polymerase chain reaction was performed for HRV and SARS-CoV-2 detection. Results: Forty-seven out of 175 (26.9%) COVID-19 adult cases have been diagnosed with asthma. The number of asthma exacerbations among the study participants was higher during 2021 than in 2020 (p=0.035). Most of the included asthmatic participants (61.7%) were only positive for SARS-CoV-2 and 38.3% were co-infected with HRV. The SARS-CoV-2 cycle threshold value was lower in samples infected with both viruses compared to samples infected with SARS-CoV-2 alone, p<0.005. Conclusion: Our findings indicate that HRV and SARS-CoV-2 were significantly more prevalent in asthma exacerbations than stable asthma. Thus, HRV and/or SARS-CoV-2 infections were potentially cofactors or contributors to the asthma exacerbation in this cohort. This is the first study, in Jordan, to investigate the HRV co-infection in COVID-19 asthmatic patients and HRV could be related with a higher severity of COVID-19.

No association between LDL receptor and CETP genetic variants and atorvastatin response in Jordanian hyperlipidemic patients.

OBJECTIVES: Atorvastatin is commonly used medication to achieve low levels of low-density lipoproteins (LDL). Cholesteryl ester transfer protein (CETP) and LDL receptor (LDLR) genetic variants can affect the cholesterol transport and hence may affect on atorvastatin response. This study aimed to investigate the influence of LDLR AvaII, CETP TaqIb, and Rs1532624 on the efficacy of 20 mg atorvastatin among Jordanian hyperlipidemic patients. METHODS: One hundred and 50 blood samples were collected from hyperlipidemic patients in the University of Jordan Hospital. Polymerase chain reaction-restriction fragment length polymorphism was used for genotyping of LDLR AvaII and CETP TaqIb genetic variants. The genotyping of CETP Rs1532624 variant was done by Sanger DNA-Sequencing. RESULTS: LDLR AvaII and CETP TaqIb and Rs1532624 variants showed a significant (p value < 0.05) association with the baseline of the LDL at the time of diagnoses. On the other hand, none of the tested genetic variants showed a significant (p value>0.05) association with LDL reduction after atorvastatin therapy. CONCLUSIONS: Results demonstrated a significant association between the LDLR AvaII and CETP TaqIb, and Rs1532624 genetic variants with the LDL baseline level. However, the atorvastatin therapy among hyperlipidemic patients of Jordanian origin was not affected by any of the tested variants.

Investigating the human rhinovirus co-infection in patients with asthma exacerbations and COVID-19

Objective: To investigate the co-infections with human rhinovirus (HRV) among patients with asthma exacerbation and COVID-19 in Jordan. Also, to determine the frequency of acute asthma exacerbation before and during the COVID-19 pandemic on a matched basis. Methods: The data of this prospective cohort research consisted of clinical variables. During the first visit, and after 14-days, nasopharyngeal swabs were taken and the quantitative polymerase chain reaction was performed for HRV and SARS-CoV-2 detection. Results: Forty-seven out of 175 (26.9%) COVID-19 adult cases have been diagnosed with asthma. The number of asthma exacerbations among the study participants was higher during 2021 than in 2020 (p=0.035). Most of the included asthmatic participants (61.7%) were only positive for SARS-CoV-2 and 38.3% were co-infected with HRV. The SARS-CoV-2 cycle threshold value was lower in samples infected with both viruses compared to samples infected with SARS-CoV-2 alone, p<0.005. Conclusion: Our findings indicate that HRV and SARS-CoV-2 were significantly more prevalent in asthma exacerbations than stable asthma. Thus, HRV and/or SARS-CoV-2 infections were potentially cofactors or contributors to the asthma exacerbation in this cohort. This is the first study, in Jordan, to investigate the HRV co-infection in COVID-19 asthmatic patients and HRV could be related with a higher severity of COVID-19. (AU)