Methyltransferase Contingencies in the Pathway of Everninomicin D Antibiotics and Analogues

Everninomicins are orthoester oligosaccharide antibiotics with potent activity against multidrug-resistant bacterial pathogens. Everninomicins act by disrupting ribosomal assembly in a distinct region in comparison to clinically prescribed drugs. We employed microporous intergeneric conjugation with Escherichia coli to manipulate Micromonospora for targeted gene-replacement studies of multiple putative methyltransferases across the octasaccharide scaffold of everninomicin effecting the A₁, C, F, and H rings. Analyses of gene-replacement and genetic complementation mutants established the mutability of the everninomicin scaffold through the generation of 12 previously unreported analogues and, together with previous results, permitted assignment of the ten methyltransferases required for everninomicin biosynthesis. The in vitro activity of A₁- and H-ring-modifying methyltransferases demonstrated the ability to catalyze late-stage modification of the scaffold on an A₁-ring phenol and H-ring C-4’ hydroxy moiety. Together these results establish the potential of the everninomicin scaffold for modification through mutagenesis and in vitro modification of advanced biosynthetic intermediates.     

Retinal Vessel Extraction Framework Using Modified Adaboost Extreme Learning Machine

An explicit extraction of the retinal vessel is a standout amongst the most significant errands in the field of medical imaging to analyze both the ophthalmological infections, for example, Glaucoma, Diabetic Retinopathy (DR), Retinopathy of Prematurity (ROP), Age-Related Macular Degeneration (AMD) as well as non retinal sickness such as stroke, hypertension and cardiovascular diseases. The state of the retinal vasculature is a significant indicative element in the field of ophthalmology. Retinal vessel extraction in fundus imaging is a difficult task because of varying size vessels, moderately low distinction, and presence of pathologies such as hemorrhages, microaneurysms etc. Manual vessel extraction is a challenging task due to the complicated nature of the retinal vessel structure, which also needs strong skill set and training. In this paper, a supervised technique for blood vessel extraction in retinal images using Modified Adaboost Extreme Learning Machine (MAD-ELM) is proposed. Firstly, the fundus image preprocessing is done for contrast enhancement and in-homogeneity correction. Then, a set of core features is extracted, and the best features are selected using “minimal Redundancy-maximum Relevance (mRmR).” Later, using MAD-ELM method vessels and non vessels are classified. DRIVE and DR-HAGIS datasets are used for the evaluation of the proposed method. The algorithm’s performance is assessed based on accuracy, sensitivity and specificity. The proposed technique attains accuracy of 0.9619 on the DRIVE database and 0.9519 on DR-HAGIS database, which contains pathological images. Our results show that, in addition to healthy retinal images, the proposed method performs well in extracting blood vessels from pathological images and is therefore comparable with state of the art methods.     

Histone Methylation Marks on Circulating Nucleosomes as Novel Blood-Based Biomarker in Colorectal Cancer

Circulating nucleic acids (CNAs) are under investigation as a liquid biopsy in cancer as potential non-invasive biomarkers, as stable structure in circulation nucleosomes could be valuable sources for detection of cancer-specific alterations in histone modifications. Our interest is in histone methylation marks with a focus on colorectal cancer, one of the leading cancers respective the incidence and mortality. Our previous work included the analysis of trimethylations of lysine 9 on histone 3 (H3K9me3) and of lysine 20 on histone 4 (H4K20me3) by chromatin immuno- precipitation-related PCR in circulating nucleosomes. Here we asked whether global immunologic measurement of histone marks in circulation could be a suitable approach to show their potential as biomarkers. In addition to H3K9me3 and H4K20me3 we also measured H3K27me3 in plasma samples from CRC patients (n = 63) and cancer free individuals (n = 40) by ELISA-based methylation assays. Our results show that of three marks, the amounts of H3K27me3 (p = 0.04) and H4K20me3 (p < 0.001) were significantly lower in CRC patients than in healthy controls. For H3K9me3 similar amounts were measured in both groups. Areas under the curve (AUC) in receiver operating characteristic (ROC) curves indicating the power of CRC detection were 0.620 for H3K27me3, 0.715 for H4K20me3 and 0.769 for the combination of both markers. In conclusion, findings of this preliminary study reveal the potential of blood-based detection of CRC by quantification of histone methylation marks and the additive effect of the marker combination.