A novel Golgi mannosidase inhibitor: Molecular design,synthesis, enzyme inhibition,and inhibition of spheroid formation

Effective chemotherapy for solid cancers is challenging due to a limitation in permeation that prevents anticancer drugs from reaching the center of the tumor, therefore unable to limit cancer cell growth. To circumvent this issue, we planned to apply the drugs directly at the center by first collapsing the outer structure. For this, we focused on cell–cell communication (CCC) between N-glycans and proteins at the tumor cell surface. Mature N-glycans establish CCC; however, CCC is hindered when numerous immature N-glycans are present at the cell surface. Inhibition of Golgi mannosidases (GMs) results in the transport of immature N-glycans to the cell surface. This can be employed to disrupt CCC. Here, we describe the molecular design and synthesis of an improved GM inhibitor with a non-sugar mimic scaffold that was screened from a compound library. The synthesized compounds were tested for enzyme inhibition ability and inhibition of spheroid formation using cell-based methods. Most of the compounds designed and synthesized exhibited GM inhibition at the cellular level. Of those, AR524 had higher inhibitory activity than a known GM inhibitor, kifunensine. Moreover, AR524 inhibited spheroid formation of human malignant cells at low concentration (10 µM), based on the disruption of CCC by GM inhibition.     

Comparative HILIC/ESI-QTOF-MS and HPTLC studies of pyrrolizidine alkaloids in flowers of Tussilago farfara and roots of Arnebia euchroma

The utility of HPTLC and HILIC/ESI-QTOF-MS for the determination of pyrrolizidine alkaloids (PAs) and their N-oxides (PANOs) was compared in the selected plant species: Tussilago farfara L. (TF, flower) and Arnebia euchroma (Royle) I.M. Johnst. (AE, root). HPTLC confirmed the postulated presence of PAs (saturated and unsaturated) or PANOs in the tested extracts. In accordance with previous studies, HILIC/ESI-Q-TOF-MS confirmed the presence of the toxic PA senkirkine and the saturated otonecine-type PAs, tussilagine and isotussilagine in the TF extract and 7-angeloylretronecine and 9-angeloylretronecine in AE extract. Moreover, the following alkaloids were identified in AE root: intermedine, intermedine-N-oxide, leptanthine-N-oxide, echimidine-N-oxide (or their corresponding stereoisomers) and traces of 7-angeloylretronecine and 9-angeloylretronecine-N-oxide. The study demonstrates the HILIC/ESI-Q-TOF-MS method to be a very useful tool for monitoring PAs and PANOs in the test samples, even when not all of the necessary standards are available. Quantitative analysis of senkirkine in TF flower by HILIC/ESI-QTOF-MS featured high resolution, high precision, high mass accuracy, and very high sensitivity with limit-of-detection (LOD) of 27.50 fg/μL and limit-of-quantitation (LOQ) of 91.60 fg/μL. The results from both methods may be used for the development or rejection of European Pharmacopoeia (X) monographs of both investigated species.     

Accuracies in Po-210 determination for lead-210 dating

Various laboratory techniques have been utilized worldwide for measuring lead-210 in sub-recent deposits through its grand-daughter product polonium-210. Isotope dilution alpha spectrometry proved a suitable tool for absolute determination of lead-210 for the dating of aquatic deposits. Moreover, isotope dilution alpha spectrometry along with speciation experiments can be used to resolve depositional anomalies arising from supported lead-210/Ra-226 disequilibrium levels and unsupported lead-210 mobile fractions. Isotope dilution alpha spectrometry of sub-recent sediment and peat deposits has been critically evaluated for more than ten years. Our results show that type, size and composition of deposits analyzed as well as radiochemical procedures used, together with alpha counting techniques, are important factors influencing lead-210 determinations and tailing corrections using its granddaughter product polonium-210. Optimization of these parameters is of prime importance to achieve economic and accurate analyses, especially at low lead-210 concentrations and small sample sizes.     

Linkage between the loci for serum albumin and vitamin D binding protein (GC) in the Japanese quail

Vitamin D binding protein ( GC ) and serum protease inhibitor ( PI ) have been added to genetic markers in the Japanese quail. Both loci were controlled by autosomal codominant alleles named GC^A, GC^B and PI^A, PI^B, PI^C, respectively. Close linkage between the loci for serum albumin ( ALB ) and GC protein is reported. Two recombinants were observed in 145 informative offspring of 14 families. The recombination frequency between the loci was estimated as 0.014±0.006. Thus, GC was assigned to linkage group II in the Japanese quail. No signs of linkage were observed among the loci for the ALB-GC complex, PI. serum prealbumin 2 ( PA2 ), phosphoglucose isomerase ( PG1 ), 6-phosphogluconate dehydrogenase ( PGD ) and esterase-D ( ESD ).     

Automatic detection of pulmonary nodules in CT images by incorporating 3D tensor filtering with local image feature analysis

Computer-aided detection (CAD) technology has been developed and demonstrated its potential to assist radiologists in detecting pulmonary nodules especially at an early stage. In this paper, we present a novel scheme for automatic detection of pulmonary nodules in CT images based on a 3D tensor filtering algorithm and local image feature analysis. We first apply a series of preprocessing steps to segment the lung volume and generate the isotropic volumetric CT data. Next, a unique 3D tensor filtering approach and local image feature analysis are used to detect nodule candidates. A 3D level set segmentation method is used to correct and refine the boundaries of nodule candidates subsequently. Then, we extract the features of the detected candidates and select the optimal features by using a CFS (Correlation Feature Selection) subset evaluator attribute selection method. Finally, a random forest classifier is trained to classify the detected candidates. The performance of this CAD scheme is validated using two datasets namely, the LUNA16 (Lung Nodule Analysis 2016) database and the ANODE09 (Automatic Nodule Detection 2009) database. By applying a 10-fold cross-validation method, the CAD scheme yielded a sensitivity of 79.3% at an average of 4 false positive detections per scan (FP/Scan) for the former dataset, and a sensitivity of 84.62% and 2.8 FP/Scan for the latter dataset, respectively. Our detection results show that the use of 3D tensor filtering algorithm combined with local image feature analysis constitutes an effective approach to detect pulmonary nodules.     

Humulus lupulus L. cv. Cascade grown in Northern Italy: morphological and phytochemical characterization

Abstract

Several aroma hops (Humulus lupulus L.) were recently introduced in Northern Italy as a small-scale production of excellence. In this preliminary study, the American cultivar Cascade was investigated in a combined morphological and phytochemical survey. Morphological investigation on trichome structure, density and distribution was performed by scanning electron microscopy and light microscopy. Essential oil composition, α/β-acid and polyphenol profiles over 3 years were determined by GC-MS and HPLC analyses. Two types of non-glandular (simple and cystolithic) and glandular (peltate and bulbous) trichomes were observed on leaves and female inflorescences. The peltate trichomes resulted as the main sites of terpene production and accumulation. The essential oil profiles showed myrcene, β-caryophyllene, (E)-β-farnesene and humulene epoxide II as the dominant compounds over the three collection times, although with different relative abundances. The presence of two exclusive compounds, γ-muurolene and trans-γ-cadinene, characterized the investigated cv. Cascade, potentially enhancing herbal, woody and spicy aroma traits of this cultivation in Northern Italy. The bitter acid composition showed quantitative values consistent with literature data only for the second and third monitoring year. Qualitative differences in polyphenol content were also recorded, for the presence of quercetin-3-O-malonylglucoside and kaempferol-3-O-rutinoside, which may characterize this peculiar Italian cultivation.     

To share or not to share: Incentivizing data sharing in life science communities

Most scientists recognize the importance of sharing data online in an open fashion. Nonetheless, many studies have documented the concerns that accompany data sharing activities, including loss of credit or IP, misuse and the time needed to curate interoperable data. To this end, discussions around data sharing often identify incentives that could potentially ameliorate these disincentivising concerns. Nonetheless, current Open Data discussions often rely on evidence‐based studies to identify the disincentives to overcome. This results in highly specific and directed interventions. In contrast, this paper offers a different interpretation of these concerns. To do so, it makes use of the Thomas Theorem which suggests that: “If men define situations as real, they are real in their consequences”. Using empirical evidence from sub‐Saharan African (bio)chemistry laboratories, this paper illustrates how individual perceptions of research environments – whether associated with evidence or not – are highly influential in shaping data sharing practices. It concludes with the suggestion that discussion on incentivising data sharing amongst scientific communities need to take a broader set of concerns into account and offer a more creative approach to ameliorating environmental disincentives.     

Adipose triglyceride lipase regulates eicosanoid production in activated human mast cells

Human mast cells (MCs) contain TG-rich cytoplasmic lipid droplets (LDs) with high arachidonic acid (AA) content. Here, we investigated the functional role of adipose TG lipase (ATGL) in TG hydrolysis and the ensuing release of AA as substrate for eicosanoid generation by activated human primary MCs in culture. Silencing of ATGL in MCs by siRNAs induced the accumulation of neutral lipids in LDs. IgE-dependent activation of MCs triggered the secretion of the two major eicosanoids, prostaglandin D2 (PGD2) and leukotriene C4 (LTC4). The immediate release of PGD2 from the activated MCs was solely dependent on cyclooxygenase (COX) 1, while during the delayed phase of lipid mediator production, the inducible COX-2 also contributed to its release. Importantly, when ATGL-silenced MCs were activated, the secretion of both PGD2 and LTC4 was significantly reduced. Interestingly, the inhibitory effect on the release of LTC4 was even more pronounced in ATGL-silenced MCs than in cytosolic phospholipase A₂-silenced MCs. These data show that ATGL hydrolyzes AA-containing TGs present in human MC LDs and define ATGL as a novel regulator of the substrate availability of AA for eicosanoid generation upon MC activation.