TLC and HPTLC Finger Printing Analysis of Cyperus rotundus (Linn.)

Cyperus rotundus (Linn.) is a versatile plant belonging to the family Cyperaceae, used in herbal medicines worldwide to cure various human ailments. The present study attempts to analyze the profiles of flavonoids in different extracts of C. rotundus with the help of thin-layer chromatography (TLC) and high-performance thin-layer chromatographic (HPTLC) fingerprint. Flower and stem extracts of C. rotundus were screens out with the help of TLC, and the Rf values were determined. HPTLC was used to quantify the flavonoid from flower extract of a plant at a 1.0 mg/mL concentration. It revealed the occurrence of flavonoids, especially quercetin in the ethanolic extract of C. rotundus flower, by using mobile phase toluene-ethyl acetate-formic acid (3:4:2.5 v/v), on a pre-coated plate of silica gel and quantified the amount of quercetin by densitometry absorbance mode at 257 nm. The limits of detection and quantification were 30.08 & 91.16 ng/mL, and the relative standard deviation ranged between 1.03 to 1.48 for intra-day and inter-day for HPTLC. The calibration range was 200-700 ng per band (r2 = 0.99321). Quercetin quantity in the ethanolic extract of the flower was found to be 0.011 mg/mL of the extract with an average recovery of 99.01–100.00%. Such fingerprinting is valuable in quality control and checking adulterants of natural drugs. Therefore, it can be helpful for the assessment of different marketable pharmaceuticals preparations.

Reinvestigation of Carbohydrate Specificity of EBCA-1 Monoclonal Antibody Used for the Detection of Candida Mannan

Monoclonal antibody EBCA-1 is used in the sandwich immune assay for the detection of circulating Candida mannan in blood sera samples for the diagnosis of invasive candidiasis. To reinvestigate carbohydrate specificity of EBCA-1, a panel of biotinylated oligosaccharides structurally related to distinct fragments of Candida mannan were loaded onto a streptavidin-coated plate to form a glycoarray. Its use demonstrated that EBCA-1 recognizes the trisaccharide β-Man-(1→2)-α-Man-(1→2)-α-Man and not homo-α-(1→2)-linked pentamannoside, as was reported previously.

Influence of Polymer Coatings on the Mechanical Properties of Steel Samples in Tensile and Bending Tests

: An experimental and theoretical study of the effect of polymer coatings on an epoxy-polyester base on the mechanical properties of samples in the form of steel plates has been carried out. It is shown that, despite the fact that the thickness of the coatings is only 100 μm, they have a significant effect on the mechanical properties of plates up to 1,5 mm thick, leading to a decrease in Young's modulus, tensile strength and ultimate deformations of the samples. It was shown that the elastic modulus of the coated plate cannot be determined unambiguously from tests for central tension and three-point bending. In bending tests, there is a more significant reduction in plate stiffness compared to tensile tests. This effect is confirmed by calculations within the framework of classical models of the theory of elasticity.

Heparanase Modulation by Wingless/INT (Wnt)

Heparanase is an endo-beta-glucuronidase, the only enzyme in mammals capable of cleaving heparan sulfate/heparin chains from proteoglycans. The oligosaccharides generated by heparanase present extensive biological functions since such oligosaccharides interact with adhesion molecules, growth factors, angiogenic factors and cytokines, modulating cell proliferation, migration, inflammation, and carcinogenesis. However, the regulation of heparanase activity is not fully understood. It is known that heparanase is synthesized as an inactive 65 kDa isoform and that post-translation processing forms an active 50 kDa enzyme. In the present study, we are interested in investigating whether heparinase is regulated by its own substrate as observed with many other enzymes. Wild-type Chinese hamster (Cricetulus griséus) ovary cells (CHO-K1) were treated with different doses of heparin. Heparanase expression was analyzed by Real-time PCR and flow cytometry. Also, heparanase activity was measured. The heparanase activity assay was performed using a coated plate with biotinylated heparan sulfate. In the present assay, a competitive heparin inhibition scenario was set aside. Exogenous heparin trigged a cell signaling pathway that increased heparanase mRNA and protein levels. The Wnt/beta-catenin pathway, judged by TCF-driven luciferase activity, seems to be involved to enhance heparanase profile during treatment with exogenous heparin. Lithium chloride treatment, an activator of the Wnt/beta-catenin pathway, confirmed such mechanism of transduction in vivo using zebrafish embryos and in vitro using CHO-K1 cells. Taken together the results suggest that heparin modulates heparanase expression by Wnt/beta-catenin.

A New Silicon Dioxide-Coated MALDI-ToF Sample Plate for Peptide Analysis

In this report, we describe the development and testing of a new coated plate which improves the sensitivity and accuracy in matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-ToF-MS). The coated plate was covered with a thin layer of hydrophobic silicon dioxide, which enabled sample enrichment due to the water repellent nature of the silicon dioxide surface. Sensitivity and required laser strengths were tested using peptide standards, with the results that these coated plates required lower laser power and showed increased sensitivity than that of common plates. Accuracy was tested using bacteria, saliva, and serum samples. The coated plates showed significantly increased degrees of accuracy through their capacity to reduce mass shift. The importance and necessity of accuracy analysis in the assessment of new sample plates, which is rarely described in other papers, is also discussed.

Smilax glabra Roxb. Inhibits Collagen Induced Adhesion and Migration of PC3 and LNCaP Prostate Cancer Cells through the Inhibition of Beta 1 Integrin Expression

Smilax glabra Roxb. (SGR) has been used as a traditional medicine for brucellosis and syphilis. In this study, we investigated whether nontoxicological levels of water extract of SGR (WESGR) are effective for suppressing steps in the progression of prostate cancer, such as collagen-mediated migration and adhesion and identified the target molecule responsible for such effects. We found that nontoxicological levels of WESGR did not attenuate PC3 and LNCaP cell adhesion to serum but did significantly do so with collagen. In addition, using the Boyden chamber assay, we found that nontoxicological levels of WESGR did not inhibit the migration of PC3 and LNCaP cells to a serum-coated area but did significantly attenuate migration to a collagen-coated area. Interestingly, the expression of α2β1 integrin, a known receptor of collagen, was not affected by ectopic administration of WESGR. However, WESGR significantly attenuated the expression of β1 integrin, but not α2 integrin when PC3 and LNCaP cells were placed on a collagen-coated plate, resulting in attenuation of focal adherent kinase phosphorylation. Finally, 5-O-caffeoylquinic acid was determined as a functional single component which is responsible for antiprostate cancer effects of WESGR. Taken together, our results suggest a novel molecular mechanism for WESGR-mediated antiprostate cancer effects at particular steps such as with migration and adhesion to collagen, and it could provide the possibility of therapeutic use of WESGR against prostate cancer progression.

Serological and Molecular Characterisation of Cucumber mosaic virus Infecting Lagenaria siceraria L. in Adim, Biase L.G.A., Cross River State, Nigeria

Cucumber mosaic virus (CMV) is one of the most important viral pathogens infecting a wide range of plant species in Nigeria. Mosaic and mottle symptoms were observed on Lagenaria siceraria L. in Adim Southern-Nigeria in 2018 and the infected leaves collected for investigation. This research was aimed at characterising the virus responsible for this infection with a view to identifying it. Antigen coated plate (ACP) enzyme linked-immunosorbent assay (ELISA) and gene sequencing were employed methods in the characterisation process. The Amplified Complementary Deoxyribonucleic Acid (cDNA) was cloned and the nucleotide sequence was determined. Result of serology revealed that the virus belonged to the genus Cucumovirus while the gene sequence obtained when compared to known virus sequences present in the GenBank using the Basic Local Alignment Search Tool (BLAST) program available at National Centre for Biotechnology Information (NCBI) revealed 97% sequence homologue with Cucumber mosaic virus confirming it as Cucumber mosaic virus. This is the first report of CMV infecting L. siceraria in Nigeria. I recommend further studies on insect and host range test of this virus be carried on.