A neutralizing antibody against human DNA polymerase epsilon inhibits cellular but not SV40 DNA replication.

The contribution of human DNA polymerase epsilon to nuclear DNA replication was studied. Antibody K18 that specifically inhibits DNA polymerase activity of human DNA polymerase epsilon in vitro significantly inhibits DNA synthesis both when microinjected into nuclei of exponentially growing human fibroblasts and in isolated HeLa cell nuclei. The capability of this neutralizing antibody to inhibit DNA synthesis in cells is comparable to that of monoclonal antibody SJK-132-20 against DNA polymerase alpha. Contrary to the antibody against DNA polymerase alpha, antibody K18 against DNA polymerase epsilon did not inhibit SV40 DNA replication in vitro. These results indicate that DNA polymerase epsilon plays a role in replicative DNA synthesis in proliferating human cells like DNA polymerase alpha, and that this role for DNA polymerase epsilon cannot be modeled by SV40 DNA replication.     

O6-Methylguanine-DNA Methyltransferase and ATP-Binding Cassette Membrane Transporter G2 Promotor Methylation: Can Predict the Response to Chemotherapy in Advanced Breast Cancer?

Background:ATP-binding cassette membrane transporter G2 (ABCG2) gene is one of transporter family and well characterized for their association with chemoresistance. Promoter methylation is a mechanism for regulation of gene expression. O6-Methyl guanine DNA methyl transferase (MGMT) gene plays a fundamental role in DNA repair. MGMT has the ability to remove alkyl adducts from DNA at the O6 position of guanine. Alkylating agents exert their function through adding these alkyls adducts to DNA leading to cell death unless it is repaired by MGMT. MGMT promoter was found to be methylated in several malignancies. The aim of the present work is to study the relation of MGMT and ABCG2 promoter methylation status in advanced breast cancer patients to response to cyclophosphamide–doxorubicin (AC) based therapeutic regime.Methods:This retrospective study included Forty-two female patients with advanced breast cancer assessed before receiving chemotherapy and after the completion of regimens. They were grouped into responders and non-responders according to RECIST criteria. Methylation analysis of MGMT and ABCG2 genes were performed on breast cancer tissues.Results:MGMT promoter was methylated in 40.5% of the cases. ABCG2 promoter was methylated in 14.3% of cases. There was no statistically significant association between MGMT and ABCG2 promoter methylation status and clinicopathological parameters. There was statistically significant association between methylation status of both promoters and response to AC when followed by Taxane.Conclusion:Methylation of MGMT and ABCG2 promoters combined could be a potential predictive factor for response to cyclophosphamide-doxorubicin based therapeutic regime.Key Words: ABCG2, Breast cancer, Chemoresistance, DNA methylation, MGMT     

Role of miRNA-210, miRNA-21 and miRNA-126 as diagnostic biomarkers in colorectal carcinoma: impact of HIF-1α-VEGF signaling pathway

Colorectal cancer (CRC) is a major cause of death worldwide. Novel non-invasive, high diagnostic value screening test is urgently needed to improve survival rate, treatment and prognosis. Stable, small, circulating microRNA (miRNA) offers unique opportunities for the early diagnosis of several diseases. It acts as tumor oncogenes or suppressors and involve in cell death, survival, and metastasis. Communication between miRNA and carcinogenesis is critical but it still not clear and needs further investigation. The aim of our study is to evaluate the role of miR-210, miR-21, miR-126, as non-invasive diagnostic biomarkers for screening, early detection of CRC, studying their correlation with prognostic variables, and clarifying the roles of miRNAs on HIF-1α-VEGF signaling pathway. The expression of miR-210, miR-21 and miR-126 was performed using qRT-PCR in adenocarcinoma (no?=?35), adenomas (no?=?51), and neoplasm free controls (no?=?101). Serum levels of VEGF and HIF-1α was determined by ELISA Kit. The results show that the expression of miR-210, miR-21, VEGF, HIF-1α was significantly up-regulated while that miRNA-126 was down-regulated in both adenocarcinoma and adenomas compared with controls (p?<?0.001 for each). No significant difference was noted comparing patients with adenocarcinoma and adenomas. The three miRNAs correlated with VEGF, HIF-α. The miR-210 and miR-21 associated with TNM classification and clinical staging of adenocarcinoma (p?<?0.001) and they show high diagnostic value with sensitivity and specificity 88.6%, 90.1% and 91.4%, 95.0% respectively. Our study revealed that circulating miR-210, miR-21 were up-regulated while miR-126 was down-regulated in CRC and adenomas patients, they all correlated with TNM staging and they had high diagnostic value. HIF-1α VEGF signaling pathways regulated by miRNAs played a role in colon cancer initiation. To the best of our knowledge, this is the first study of this miRNAs panel in CRC in our community. These data suggested that these biomarkers could be a potential novel, non-invasive marker for early diagnosis, screening and predicting prognosis of CRC. Understanding the molecular functions by which miRNAs affect cancer and understanding its roles in modulating the signaling output of VEGF might be fruitful in reducing the incidence and slowing the progression of this dark malignancy.

     

Performance of some elite potato cultivars under abiotic stress at North Sinai

The current experiment was adopted during the summer 2018, fall 2018/2019 and summer 2019 respectively at the Experimental Farm of Baloza station, Desert Research Center. North Sinai Governorate, Egypt to study the effect of different doses of irradiation (0, 20, 30 and 40 Gy), three irrigation levels (100, 80 and 60% field capacity on growth, yield and its quality of some potato cultivars (Spunta, Cara, Caruso and Hermes). Treated Spunta cultivar pre planting with 20 (Gy) and irrigated with 80% field capacity was the best treatment for increasing number of aerial stem/plants, leaf area, total chlorophyll in leaves, average tuber weight, and total yield/fed. Hermes cultivar with 20 (Gy) and irrigation level of 80% was the best for increasing dry matter content in tuber in both mutagenic generations.     

Synthesis and biological evaluation of 2-styrylquinolines as antitumour agents and EGFR kinase inhibitors: molecular docking study

A new series of 4,6-disubstituted 2-(4-(dimethylamino)styryl)quinoline 4a,b–9a,b was synthesized by the reaction of 2-(4-(dimethylamino)styryl)-6-substituted quinoline-4-carboxylic acids 3a,b with thiosemicarbazide, p-hydroxybenzaldehyde, ethylcyanoacetate, and 2,4-pentandione. In addition, the antitumour activity of all synthesized compounds 3a,b–9a,b was studied via MTT assay against two cancer cell lines (HepG2 and HCT116). Furthermore, epidermal growth factor receptor (EGFR) inhibition, using the most potent antitumour compounds, 3a, 3b, 4a, 4b, and 8a, was evaluated. The interpretation of the results showed clearly that the derivatives 3a, 4a, and 4b exhibited the highest antitumour activities against the tested cell lines HepG2 and HCT116 with IC₅₀ range of 7.7–14.2?µg/ml, in comparison with the reference drugs 5-fluorouracil (IC₅₀?=?7.9 and 5.3?µg/ml, respectively) and afatinib (IC₅₀?=?5.4 and 11.4?µg/ml, respectively). In vitro EGFR screening showed that compounds 3a, 3b, 4a, 4b, and 8a exhibited moderate inhibition towards EGFR with IC₅₀ values at micromolar levels (IC₅₀ range of 16.01–1.11?µM) compared with the reference drugs sorafenib (IC₅₀ =?1.14?µM) and erlotinib (IC₅₀ =?0.1?µM). Molecular docking was performed to study the mode of interaction of compounds 3a and 4b with EGFR kinase.     

Synthesis,Characterization, and Biological Evaluation of Cu(II) Complexes Containing Triflupromazine with Glycine and Histidine

Two novel copper (II) complexes [Cu(TFP)(Gly)Cl] ⋅ 2H₂O complex ( 1 ) and [Cu(TFP)(His)Cl] ⋅ 2H₂O complex ( 2 ) are synthesized, where TFP stands for trifluropromazine, Gly. represents glycine, and His. is histidine. Chemical composition, IR, mass spectra, and magnetic susceptibility tests are performed. Complex binding with macromolecules was investigated using UV-vis, viscosity, gel electrophoresis, and fluorescence quenching. Fluorescence spectroscopy revealed that each complex could replace ethidium bromide (EB). These complexes exhibit grooved, non-covalent, and electrostatic interactions with CT-DNA. Spectroscopy analysis of the BSA interaction showed that complexes bind to protein (K_b values for ( 1 ) is 5.89×10³ M⁻¹ and for ( 2 ) is 9.08×10³ M⁻¹) more strongly than CT-DNA (K_b values for ( 1 ) is 5.43×10³ M⁻¹ and for ( 2 ) is 7.17×10³ M⁻¹). Molecular docking analysis and spectral absorption measurements showed high agreement. Antimicrobial, antioxidant, and anti-inflammatory properties were tested in vitro. The druggability of complex ( 2 ) should be tested in vivo as it is more biologically active.     

Multivariate analysis of vegetational variation in different habitats at Omayed,Egypt

The application of indicator species analysis has proved useful in classifying stands into groupings coinciding with topographic variations. 41 indicator pseudo-species are identified in the vegetation along a phytosociological gradient: some are characteristic of specific habitats, and the others indicate transition between groups of habitats.The use of qualitative estimations (transformed density records) in multivariate analysis in the present study indicates that they may be more preferable than quantitative estimations, because of their easy and fast recording in the field, and the less comprehensive computations, while yielding precise results.The X-axis of the reciprocal averaging ordination is related to the salinity and fertility gradients, while the Y-axis reflects soil texture.The phytosociological gradients of the canonical variate axes reflect essentially contrasts between groups of species, each correlated with one or the other of the environmental gradients.Comparing the results obtained by the two ordination methods (reciprocal averaging and canonical variates) we find that the first method detects the overall phytosociological changes along strong environmental gradients which would be helpful in studying large surveys. On the other hand, the second technique, which is mainly predictive, is more sensitive to changes within both the phytosociological and environmental gradients and can detect the impact of these changes on the overall variance of each gradient.Nomenclature follows Täckholm (1964).The authors are grateful to Dr J. Jeffers and Mr and Mrs P. Howard of the Institute of Terrestrial Ecology, England, for their help with applying the multivariate techniques to the vegetation data.     

A newEuphorbia species from Southern Sinai

A new species,Euphorbia sanctae-catharinae A. Fayed, from the group ofE. terracina L., is described from southern mountaneous area of Sinai.