Biochemical and molecular study on interleukin-1ß gene expression and relation of single nucleotide polymorphism in promoter region with Type 2 diabetes mellitus.

Interleukin-1ß (IL-1ß) assumes a centric role in the regulation of immune and inflammatory responses and thus has been recognized in immune mediated diseases like type 2 diabetes mellitus (T2DM). We aimed to investigate expressed level of IL-1ß and its relation with IL-1ß -511T>C polymorphism in T2DM patients. This study enrolled 80 subjects (50 patients with T2DM and 30 healthy control subjects). Laboratory investigations included fasting (FBG) and 2 h postprandial blood sugar (2 h PBG), HBA1c, lipid profile, and renal function tests. Genotyping of IL-1ß -511T>C (rs16944) SNP assay by real-time PCR and relative quantitation of IL-1ß gene expression transcript by real-time PCR. RESULTS: T2DM patients had significantly higher FBG and 2 h PBG, HBA1c, LDLc, TC, TG, systolic, and diastolic BP while lower HDLc compared with control group. IL 1- ß -511 T>C, CC genotype and C allele were significantly associated with risk of T2DM with odds ratio (OR) 4.73, 95%CI (1.21-18.39) and OR 2.27, 95%CI (1.72-4.40), respectively. Moreover, diabetic patients had significantly higher IL 1- ß gene transcript compared with control group (P < 0.001). CC genotype of IL 1- ß -511 T > C had the highest significant level of IL 1- ß gene transcript demonstrated compared with C/T and T/T genotypes (P < 0.001) in patients. CONCLUSION: C allele of IL-1 ß -511 T >C could be considered risk factor contributor to T2DM and excess level of IL-1 ß transcript may disclose to some degree the inflammatory role of cytokines in T2DM.

Potential role of MicroRNA 200c gene expression in assessment of colorectal cancer.

BACKGROUND AND AIM: Colorectal cancer (CRC) is a common cancer worldwide that affects men and women of all racial and ethnic groups. Recent evidence supports the role of microRNAs in CRC. We planned to investigate microRNA200c expression and its relation with diagnosis, prognosis, metastasis and overall survival in CRC patients. This study enrolled 90 subjects (3'0 CRC patients, 30 patients with benign colorectal polyps and 30 healthy control subjects). METHODS: Laboratory investigations included measurement of serum CA19-9 and CEA by enzyme linked immunosorbent assay (ELISA) method and relative quantitation (RQ) of microRNA200c gene expression by real time PCR technique. RESULTS: Significant higher MicroRNA200c expression levels in CRC patients versus both benign (P < 0.011) and control groups (P < 0.001), additionally, benign group had elevated levels versus control (P < 0.001). MicroRNA 200c at cutoff >4.56 had sensitivity 86.67% and specificity 73.33% (P < 0.001) for CRC discrimination. Kaplan-Meier survival analysis revealed significant association (P = 0.028) of high expression of microRNA200c with decreased overall survival. CONCLUSION: Noticeable up-regulation of microRNA200c in CRC and its remarkable relation with unfavorable survival suggesting its potential dual use as a diagnostic and prognostic biomarker for CRC.

A preliminary study of the relation between IL-4 and hypertension in type II diabetes mellitus.

Clinical outcome of T2DM can be influenced by a polymorphism of different cytokines. The aim of this study was to evaluate the effect of genetic variation and gene expression of IL-4 in T2DM patients. This study was carried out on type II diabetic patients and healthy people served as controls. All subjects were submitted to estimation of IL-4 gene polymorphism using VNTR PCR method and gene expression by real-time PCR. There was a significant decrease of IL-4 gene expression and serum IL4 levels in subjects with B2B2 genotypes. A significant positive correlation was found between IL-4 gene expression and the HDL-c levels and negative correlation between serum IL4 levels and LDLc. Also, a negative correlation was found between serum IL4 and gene expression with both systolic and diastolic blood pressure levels in patients group. It can be concluded that IL-4 gene expression and serum IL4 reduction in patients with B2B2 genotypes has a relation to dyslipidemia and hypertension in the patient with type 2 diabetes mellitus.

Five Fibrosis Biomarkers Together with Serum Ferritin Level to Diagnose Liver Fibrosis and Cirrhosis.

BACKGROUND: Liver fibrosis is a dynamic procedure that results from an irregularity between fibrogenesis and fibrolysis. After time this procedure can lead to cirrhosis of the liver. Liver fibrosis and cirrhosis assessment is very important for both therapeutic decisions and prognostic evaluations. In this study, we tried to use serum ferritin (SF) together with five fibrosis tests (Age-Platelet index (API), aspartate aminotransferase to alanine aminotransferase ratio (AAR), AST to platelet ratio index (APRI), Fibrosis 4 score (FIB-4), and fibro-quotient (Fibro-Q)) to assess liver fibrosis and cirrhosis and estimate possible correlation between inflammation and SF. METHODS: This study was carried out on eighty-eight patients infected with HCV and twenty healthy subjects as a control. Complete blood count (CBC), aspartate aminotransferase (AST), alanine aminotransferase (ALT), antiHCV antibody, detection of HCV RNA by real-time PCR, and serum ferritin (SF) were assessed. Then API, ARR, APRI, FIB-4, and Fibro-Q were calculated. Different fibrosis stages (mild fibrosis stage (F1), moderate fibrosis stage (F2), severe fibrosis stage (F3), cirrhotic stage (F4)) were assessed using transient elastography by Fibro Scan®. RESULTS: FIB-4 index was significantly elevated (p < 0.01) with the progression of liver fibrosis at F1, F2, F3, and F4 when compared to healthy control group. The APRI score elevation between F0 and F3 and between F0 and F4 was significant (p < 0.01). SF was elevated in all fibrosis stages and significantly (p < 0.01) at F3 and F4 compared to controls. CONCLUSIONS: APRI coupled with SF should be the best reliable biomarkers for liver cirrhosis. Simultaneously, from our data SF involved in all stages of inflammation. Therefore, down regulation of ferritin in the early stage of fibrosis should be helpful in decreasing the inflammatory effect of ferritin.

Metformin, an Anthropogenic Contaminant of Seidlitzia rosmarinus Collected in a Desert Region near the Gulf of Aqaba, Sinai Peninsula.

A phytochemical investigation of Seidlitzia rosmarinus collected along the shoreline of the Gulf of Aqaba in the remote southern desert region of the Sinai peninsula has revealed the presence of the registered drug metformin (4). However, analysis of the 14C content revealed the drug to be an anthropogenic contaminant. Consequently, natural product researchers should be aware that compounds isolated from plants might originate from environmental contamination rather than biosynthesis. The new natural product N-(4-hydroxyphenylethyl)-α-chloroferuloylamide was isolated as a mixture of the E and Z isomers along with a number of other well-established secondary metabolites.

Synthesis of some new carbonitriles and pyrazole coumarin derivatives with potent antitumor and antimicrobial activities.

3-Acetyl-4-hydroxycoumarin (2) was reacted with some aldehydes (4-chlorobenzaldehyde, 4-bromobenzaldehyde, 5-methylfurfural) to afford the chalcones (3a-c). Cyclization of these chalcones with malononitrile in the presence of ammonium acetate afforded pyridine carbonitriles (4a-c), while the cyclization reaction of chalcones (3a-c) with ethyl cyanoacetate afforded the oxopyridine carbonitriles (5a-c). On the other hand, the chalcones (3a-c) reacted with hydrazine hydrate in alcohol to yield pyrazoles (6a-c), but when the same reaction is carried out in the presence of acetic acid, the acetyl pyrazole derivatives (7a-c) were obtained. Finally, the reaction of the chalcones (3a-c) with phenylhydrazine afforded phenylpyrazole derivatives (8a-c). The structures of synthesized compounds were confirmed by their micro analysis and spectral data (IR, NMR and MS). Twelve samples were evaluated for the human breast adenocarcinoma cytotoxicity, three of them showed moderate activity, the rest of the samples showed weak cytotoxic activity (very high IC50), but for the hepatocarcinoma cell lines four samples showed weak cytotoxic effect, while the rest of the compounds showed very weak effect. For antimicrobial study, three compounds proved to be the most promising against tested bacterial organisms.

MicroRNA-29a and MicroRNA-124 as novel biomarkers for hepatocellular carcinoma.

BACKGROUND: Numerous microRNAs (miRNAs) have been observed to be abnormally expressed in cancer. Therefore, miRNA signatures could be potential noninvasive diagnostic and prognostic biomarkers for hepatocellular carcinoma (HCC). AIMS: To correlate miRNA-29a and miRNA-124 expression levels with the clinical features and survival rates of HCC patients. METHODS: Serum miRNA expression in 150 samples (50 patients with HCC, 50 patients with liver cirrhosis, and 50 healthy controls) were quantified using real-time qRT-PCR. RESULTS: The expression levels of serum miRNA-29a were higher and the levels of miRNA-124 were lower in patients with HCC than in patients with liver cirrhosis and controls. ROC curve analysis showed promising accuracy for both miRNAs in distinguishing patients with HCC from those with liver cirrhosis. Levels of miRNA-29a were related to tumor number, size, stage, and outcome, whereas levels of miRNA-124 were related to vascular invasion. The overall survival rate of patients with low miRNA-29a expression was significantly higher than that of patients with high expression. Additionally, the multivariate analysis identified miRNA-29a as an independent prognostic variable. CONCLUSIONS: The investigated miRNAs showed acceptable accuracy in the diagnosis of HCC; therefore, both could be utilized as diagnostic biomarkers. Additionally, miRNA-29a could be used as a prognostic biomarker.

Down regulation of miR-30a-5p and miR-182-5p in gastric cancer: Clinical impact and survival analysis.

BACKGROUND AND AIM: Gastric Cancer (GC) is a leading cause of morbidity and mortality worldwide, particularly in developing nations, only a few suitable gastric cancer serum biomarkers with acceptable sensitivity and specificity exist. This work aims to highlight and uncover miR-30a-5p and miR-182-5p's diagnostic roles regarding gastric cancer and their roles in predicting prognosis. METHODS: 148 patients participated in this study. Groups I, II, and III had 47 patients with GC, 54 patients with benign gastric lesions, and 47 apparently healthy subjects of coincided age and gender as controls, respectively. All participants were clinically evaluated and subjected to CBC, serum CEA, and CA19-9 by ELISA, and real-time PCR tests of miR-30a-5p and miR-182-5p. RESULTS: MiR30a-5p and miR-182-5p were down regulated in gastric cancer patients in Group I more than Groups II and III (P < 0.001). ROC curve analysis revealed that miR30a-5p had better AUC, sensitivity, and specificity (0.961%, 93.62%, and 90.74%respectively). When miR-182-5p was gathered with CEA and CA19-9, specificity raised to 98.15% and PPV to 97.6%. Lower miR-30a-5p levels are linked with the presence of distant metastases, advanced TNM stage, and degree of pathological differentiation of tumors in GC patients (p = 0.034, 0.019, 0.049) respectively. According to the multivariate analysis, miR30a-5p expression level could be an independent predictor of GC. CONCLUSION: Our results exhibited that miRNAs, miR-30a-5p and miR182-5p, gene expression have a diagnostic power and can identify patients with GC. MiR-30a-5p displayed the highest diagnostic specificity and sensitivity. Besides other known tumor markers, they could offer simple noninvasive biomarkers that predict gastric cancer.

The diagnostic and prognostic role of MiRNA 15b and MiRNA 378a in neonatal sepsis.

BACKGROUND AND OBJECTIVES: Sepsis is one of the major factors for both term and preterm babies with morbidity and mortality. On the basis of recent clinical trials, altered circulating micro-RNAs (miRNAs) may serve as possible biomarkers in sepsis for diagnosis and prognosis. The aim of this research is to assess the diagnostic and prognostic biomarkers of miRNA 15b and miRNA 378a for neonatal sepsis. SUBJECTS & METHODS: This study was carried out 25 neonates with sepsis admitted to neonatal ICU of Menoufia University Hospital and 25 healthy controls from February 2019 to May 2020. The relative quantification (RQ) of miRNA-15b and miRNA-378a expression was assessed using real time PCR technique. Results: Our results demonstrated that patients with sepsis had significantly higher level of MiRNA-15b than the healthy volunteers. On the other hand, patients with sepsis had significantly lower level of MiRNA-378a than the healthy volunteers. The ROC curve showed that the serum MiRNA-15b was a significant discriminator of sepsis with a combined sensitivity and specificity of 76% and 88% with cutoff point of 3.24. In addition, serum MiRNA-378a was a significant discriminator of sepsis with a combined sensitivity and specificity of 60% and 88% with cutoff point of 0.361. The miRNA-15b expression significantly correlated positive with respiratory rate (r =0.415,p =0.039), WBCs (r = 0.408, p =0.043), and CRP (r =0.407, p=0.043). Likewise, miRNA-378a expression significantly correlated negative with respiratory rate (r =-0.415p =0.024), WBCs (r =- 0.442, p =0.027), and CRP (r =- 0.459, p=0.021). CONCLUSION: Both MiRNA 15b and 378a are promising biomarker for neonatal sepsis.

Chemical constituents from Limonium tubiflorum and their in silico evaluation as potential antiviral agents against SARS-CoV-2.

Wild plants growing in the Egyptian deserts are facing abiotic stress, which can lead to interesting & safe natural products possessing potential chemical profiles. Consequently, our study was designed to assess the phytochemical composition of the aerial parts of Limonium tubiflorum (family Plumbaginaceae) growing wild in Egypt for the first time. In addition, in silico screening and molecular dynamic simulation of all isolated phytoconstituents were run against the main protease (Mpro) and spike glycoprotein SARS-CoV-2 targets which displayed a crucial role in the replication of this virus. Our findings showed that the phytochemical investigation of 70% ethanol extract of L. tubiflorum aerial parts afforded six known flavonoids; myricetin 3-O-(2''-galloyl)-ß-d-galactopyranoside (1), myricetin 3-O-(2''-galloyl)-α-l-rhamnopyranoside (2), myricetin 3-O-(3''-galloyl)-α-l-rhamnopyranoside (3), myricetin 3-O-ß-d-galactopyranoside (5), apigenin (6), myricetin (7), along with two known phenolic acid derivatives; gallic acid (4) and ethyl gallate (8). Docking studies revealed that compounds (1) & (2) were the most effective compounds with binding energies of -17.9664 & -18.6652 kcal mol-1 against main protease and -18.9244 & -18.9272 kcal mol-1 towards spike glycoprotein receptors, respectively. The molecular dynamics simulation experiment agreed with the docking study and reported stability of compounds (1) and (2) against the selected targets which was proved by low RMSD for the tested components. Moreover, the structure-activity relationship revealed that the presence of the galloyl moiety is necessary for enhancement of the activity. Overall, the galloyl substructure of myricetin 3-O-glycoside derivatives (1 and 2) isolated from L. tubiflorum may be a possible lead for developing COVID-19 drugs. Further, in vitro and in vivo assays are recommended to support our in silico studies.

In vitro antioxidant and antiproliferative activities of flavonoids from Ailanthus excelsa (Roxb.) (Simaroubaceae) leaves.

The present study aimed to investigate the chemical composition, and the antioxidant and antiproliferative activities of Ailanthus excelsa, a plant used in Egyptian traditional medicine. Chromatographic separation of a methanol extract of A. excelsa leaves yielded four flavones, namely apigenin (1), apigenin 7-O-beta-glucoside (2), luteolin (3), and luteolin 7-O-beta-glucoside (4), and seven flavonols, namely kaempferol (5), kaempferol 3-O-alpha-arabinoside (6), kaempferol 3-O-beta-galactoside (7), quercetin (8), quercetin 3-O-alpha-arabinoside (9), quercetin 3-O-beta-galactoside (10), and quercetin 3-O-rutinoside (11). The A. excelsa extract tested in different in vitro systems (DPPH and FRAP assays) showed significant antioxidant activity. The potential antiproliferative activity of the A. excelsa extract and isolated flavonoids against five human cancer cell lines such as ACHN, COR-L23, A375, C32, and A549 was investigated in vitro by the SRB assay in comparison with one normal cell line, 142BR. The extract exhibited the highest inhibitory activity against C32 cells with an IC50 value of 36.5 microg ml(-1). Interesting activity against COR-L23 was found with 10 (IC50 value of 3.2 microg ml(-1)). Compounds 1 and 3 inhibited cell growth in both amelanotic melanoma and malignant melanoma cells.

CXCL9 chemokine level is associated with spontaneous clearance and sustained virological response in Egyptian Chronic Hepatitis C patients receiving direct acting antivirals.

BACKGROUND: Chronic Hepatitis C virus (HCV) infection is associated with progressive liver inflammation which in turn leads to cirrhosis and finally causes hepatocellular carcinoma (HCC). By different escape mechanisms, the virus succeeds to evade the innate and acquired immune responses to establish chronic infection. AIM: This study aimed to evaluate the level of chemokine CXCL9 and its correlation with some biochemical parameters in different subjects of HCV patients. MATERIALS AND METHODS: A total of 83 persons participated in this study including healthy subjects without both HCV antibodies and HCV RNA (22.9%), HCV treated responders accomplished SVR post treatment, with HCV antibodies and absence of HCV RNA (24.1%), spontaneous or natural clearance patients, with positive HCV antibodies and negative HCV RNA without treatment (26.5%) and chronic HCV-patients, with both positive HCV antibodies and HCV RNA with no treatment (26.5%). HCV RNA was quantitated by real time PCR and serum CXCL9 level was measured by ELISA commercial kit pre-coated with human MIG/CXCL9 antibody. Assessment of biochemical and hematological parameters was carried out. RESULTS: Data showed that, the level of CXCL9 was significantly increased in chronic individuals (627.1 pg/ml) (P< 0.001) than spontaneous clearance (107.76 pg/ml) and responder subjects (117.28 pg/ml) (P⩽ 0.05). No correlation has been found between CXCL9 level and viral load. Furthermore, CXCL9 levels correlated variably with some biochemical and hematological parameters according to each subject. CONCLUSION: Serum Chemokine CXCL9 level is associated with spontaneous clearance of HCV and response to HCV treatment, which may be identified as a predictive marker among HCV patients.

Ficus retusa-stabilized gold and silver nanoparticles: Controlled synthesis, spectroscopic characterization, and sensing properties.

Ficus retusa was used as reducing and stabilizing agent in the green synthesis of silver and gold nanoparticles with high dispersion stability and controllable size and shape. The controlling of reaction conditions i.e. contact time, extract quantity, metal concentration, and pH value enables the tuning of the particle size and size distribution of the metal nanoparticles. UV-visible spectroscopy was used to follow the spectral profile changes of the surface plasmon resonance of the metal nanoparticles due to different treatments. The surface plasmon resonance varies between 400 and 432 nm and between 522 and 554 nm for silver and gold nanoparticles, respectively, depending on the different reaction parameters. Atomic force and transmission electron microscopy results confirmed the success of preparation of spherical silver (15 nm) and gold (10-25 nm) nanoparticles with narrow size-distribution. Fourier transform infrared spectroscopy suggested the phenolic compounds play the key role in the reduction and stabilizing of metal ions. The colorimetric sensitivity of silver and gold nanoparticles to detect the presence of heavy metals in water was studied.

The clinical impact of miRNA34a and P53 gene expression in colon cancer.

OBJECTIVE: To study the potential role of miRNA34a gene expression and its relationship with P53 gene expression, fate, stage, metastasis and overall survival of colorectal cancer. PATIENTS AND METHODS: This study was carried out 30 patients with colon adenocarcinoma, 30 patients with benign colon polyp and 30 apparently healthy persons served as controls. All participants were subjected to full history taking, general clinical examination. Complete blood count, liver and kidney function, determination of serum tumor markers were done. Estimation of microRNA 34a and P53 Gene expression by real-time PCR were done. RESULTS: There was a significant negative relationship between serum tumor markers and micro RNA 34a gene expression in cancer patients. Also, there was a statistically significant positive relationship between miRNA34a gene expression and P53 gene expression in both patients groups. The diagnostic accuracy of miRNA34a gene expression was both sensitive and specific for colon cancer. MiRNA34a and P53 gene expression had statistically significant relation with tumor stage and presence of metastases. CONCLUSION: It can be concluded that the level of miRNA34a can be used to differentiate between colon cancers and begin adenomas. MiRNA34a can be used as a prognostic marker in colon cancer.

Design, synthesis and evaluation of novel diaryl pyrrolopyrimidine and pyrrolothiazine derivatives as inhibitors of tumor necrosis factor stimulated gene-14 (TSG-14) production.

A novel series of pyrrolothiazines 2-4 and pyrrolopyrimidines 5-7 have been synthesized. The structures of these compounds were established by spectroscopic and element microanalytical data. The newly synthesized compounds were evaluated as inhibitors of TSG-14. The most effective results were obtained by the S-sec-butyl derivatives 6e (80%) and the N-ethyl derivatives 4e (70%).

A new gorgostane derivative from the Egyptian Red Sea soft coral Heteroxenia ghardaqensis.

Chemical investigation of the chloroform extract of the Egyptian Red Sea soft coral, Heteroxenia ghardaqensis (Family Xeniidae), led to the isolation of three gorgostane derivatives, namely gorgosten-5(E)-3ß-ol (1), gorgostan-3ß,5α,6ß, 11α-tetraol (sarcoaldosterol A) (2) and gorgostan-3ß,5α,6ß-triol-11α-acetate (3). To our knowledge, sterol 3 is reported in this article for the first time. The structure elucidation of these compounds was deduced by 1-D and 2-D NMR as well as ESIMS. Sterol 1 showed moderate activity as growth inhibitor of human colon tumour cell lines.

Phenolic metabolites from Pyruscalleryana and evaluation of its free radical scavenging activity.

Investigation of the aqueous alcoholic extract of Pyruscalleryana Decne. leaves led to the isolation of two new phenolic acids glycosides, namely protocatechuoylcalleryanin-3-O-ß-glucopyranoside (1) and 3'-hydroxybenzyl-4-hydroxybenzoate-4'-O-ß-glucopyranoside (2), together with nine known compounds among them lanceoloside A and methylgallate, which have been isolated for the first time from the genus Pyrus. Structures of the isolated compounds were established by spectroscopic analysis, including UV, IR, HRESI-MS, and 1D/2D NMR. The total extract and some isolated compounds were determined against DPPH (2,2-diphenyl-1-(2,4,6-trinitrophenyl) hydrazinyl radical, for their free radical scavenging activity, the total alcoholic extract showed strong antioxidant activity while the two new compounds showed weak antioxidant activity.