GD2 ganglioside-binding antibody 14G2a and specific aurora A kinase inhibitor MK-5108 induce autophagy in IMR-32 neuroblastoma cells

The process of autophagy and its role in survival of human neuroblastoma cell cultures was studied upon addition of an anti-GD2 ganglioside (GD2) 14G2a mouse monoclonal antibody (14G2a mAb) and an aurora A kinase specific inhibitor, MK-5108. It was recently shown that combination of these agents significantly potentiates cytotoxicity against IMR-32 and CHP-134 neuroblastoma cells in vitro, as compared to the inhibitor used alone. In this study we gained mechanistic insights on autophagy in the observed cytotoxic effects exerted by both agents using cytotoxicity assays, RT-qPCR, immunoblotting, and autophagy detection methods. Enhancement of the autophagy process in the 14G2a mAb- and MK-5108-treated IMR-32 cells was documented by assessing autophagic flux. Application of a lysosomotropic agent—chloroquine (CQ) affected the 14G2a mAb- and MK-5108-stimulated autophagic flux. It is our conclusion that the 14G2a mAb (40 μg/ml) and MK-5108 inhibitor (0.1 μM) induce autophagy in IMR-32 cells. Moreover, the combinatorial treatment of IMR-32 cells with the 14G2a mAb and CQ significantly potentiates cytotoxic effect, as compared to CQ used alone. Most importantly, we showed that interfering with autophagy at its early and late step augments the 14G2a mAb-induced apoptosis, therefore we can conclude that inhibition of autophagy is the primary mechanism of the CQ-mediated sensitization to the 14G2a mAb-induced apoptosis. Although, there was no virtual stimulation of autophagy in the 14G2a mAb-treated CHP-134 neuroblastoma cells, we were able to show that PHLDA1 protein positively regulates autophagy and this process exists in a mutually exclusive manner with apoptosis in PHLDA1-silenced CHP-134 cells.     

Vascular extracellular adenosine metabolism in mice correlates with susceptibility to atherosclerosis

Abstract

Animal models are widely used in atherosclerosis research. The most useful, economic and valid is mouse genetic model of this pathology. Purinergic signaling is an important mechanism regulating processes involved in the vascular inflammation and atherosclerosis. The aim of this study was to measure vascular activities of nucleotide and adenosine-degrading ecto-enzymes in different strains of mice and to compare them to atherosclerotic susceptibility.

The vascular extracellular nucleotide catabolism pathway was analyzed in 6-month-old male genetically unmodified mouse strains: FVB/NJ, DBA/2J, BALB/c, C57Bl/6J and mouse knock-outs on C57Bl/6J background for LDLR (LDLR-/-) and for ApoE and LDLR (ApoE-/-LDLR-/-). LDLR-/- mice were a model of moderate hypercholesterolemia, while ApoE-/-LDLR-/- mice, a model of severe hypercholesterolemia with advanced atherosclerosis.

FVB/NJ, DBA/2J and BALB/c mice showed high rates of vascular extracellular AMP hydrolysis and low activity of adenosine deamination. In turn, all mice with the C57Bl/6J background expressed diminished activity of vascular AMP hydrolysis. Mice with genetically-induced hyperlipidemia and atherosclerosis on the C57Bl/6J background revealed increased ecto-adenosine deaminase activity.

Mouse strains that were resistant to atherosclerosis (FVB/NJ, DBA/2J, BALB/c) exhibited a protective extracellular vascular ecto-enzyme pattern directed toward the production of anti-inflammatory and anti-atherosclerotic adenosine. In turn, mice with genetically induced hypercholesterolemia and atherosclerosis expressed disturbed activities of ecto-5’nucleotidase and ecto-adenosine deaminase related to decreased production and increased degradation of extracellular adenosine.     

Ant communities and Solidago plant invasion: Environmental properties and food sources

The invasion of Solidago is one of the main threats to the biodiversity of natural meadows, leading to changes in animal and plant communities, as well as soil features. We compared effects of soil microclimatic conditions (temperature and moisture) and the availability of potential protein sources (dry mass of epigean invertebrates) on ants between meadows invaded by Solidago altissima and S. canadensis and those uninvaded. Our results showed that the ant communities were different between the uninvaded and invaded meadows, with reduction of ant abundance and species richness in the latter. Myrmica spp. were abundant in the uninvaded meadows, whereas Lasius niger was the dominant species in the invaded ones. We found that the lower moisture negatively influenced the abundance of Myrmica species in the Solidago‐invaded meadows. Moreover, the epigean invertebrate dry mass, as an estimation of the availability of protein sources, varied between the two types of meadows, with a higher abundance in the uninvaded ones. The abundance of Myrmica ants with narrower ecological requirements showed a positive correlation with the invertebrate biomass in the invaded meadows. In contrast, the abundance of L. niger with broad ecological requirements was negatively correlated with the invertebrate biomass in the invaded meadows, possibly as a strategy to reduce interspecific competition. Our study showed that the invasion of Solidago plants caused changes in the abundance and species composition of ant communities through modification in microhabitat conditions, that is, decreasing soil moisture, reducing biomass and changing distribution of prey invertebrates.     

Extreme weather‐year sequences have nonadditive effects on environmental nitrogen losses

The frequency and intensity of extreme weather years, characterized by abnormal precipitation and temperature, are increasing. In isolation, these years have disproportionately large effects on environmental N losses. However, the sequence of extreme weather years (e.g., wet–dry vs. dry–wet) may affect cumulative N losses. We calibrated and validated the DAYCENT ecosystem process model with a comprehensive set of biogeophysical measurements from a corn–soybean rotation managed at three N fertilizer inputs with and without a winter cover crop in Iowa, USA. Our objectives were to determine: (i) how 2‐year sequences of extreme weather affect 2‐year cumulative N losses across the crop rotation, and (ii) if N fertilizer management and the inclusion of a winter cover crop between corn and soybean mitigate the effect of extreme weather on N losses. Using historical weather (1951–2013), we created nine 2‐year scenarios with all possible combinations of the driest (“dry”), wettest (“wet”), and average (“normal”) weather years. We analyzed the effects of these scenarios following several consecutive years of relatively normal weather. Compared with the normal–normal 2‐year weather scenario, 2‐year extreme weather scenarios affected 2‐year cumulative NO₃^? leaching (range: ?93 to +290%) more than N₂O emissions (range: ?49 to +18%). The 2‐year weather scenarios had nonadditive effects on N losses: compared with the normal–normal scenario, the dry–wet sequence decreased 2‐year cumulative N₂O emissions while the wet–dry sequence increased 2‐year cumulative N₂O emissions. Although dry weather decreased NO₃^? leaching and N₂O emissions in isolation, 2‐year cumulative N losses from the wet–dry scenario were greater than the dry–wet scenario. Cover crops reduced the effects of extreme weather on NO₃^? leaching but had a lesser effect on N₂O emissions. As the frequency of extreme weather is expected to increase, these data suggest that the sequence of interannual weather patterns can be used to develop short‐term mitigation strategies that manipulate N fertilizer and crop rotation to maximize crop N uptake while reducing environmental N losses.     

Novel tool in rheumatoid arthritis diagnosis—The usage of urease flap region peptidomimetics

Rheumatoid arthritis (RA) is an autoimmune inflammatory disease. Early diagnosis can prevent joint erosion. However, available biomarkers do not always allow for clear distinction between RA and non‐RA individuals. It has become known that bacteria/viruses are among the environmental triggers that initiate RA via multiple molecular mechanisms. Thus, to better understand the role of bacteria in RA, we synthetized 6 peptidomimetics of bacterial ureases' flap region. These peptides were then used to distinguish RA patients from healthy people sera by immunoblotting. Most patients' sera were bound to peptidomimetic characteristic for Enterobacter sp. and Klebsiella sp. flap urease. We also found similarities between peptidomimetic sequence and human proteins connected with RA. This pilot study suggests that bacteria may trigger RA via mechanism of molecular mimicry of urease to host proteins and ureases flap peptidomimetics may be potential candidate as a new additional diagnostic test.     

Nanosilica synthesis mediated by Aspergillus parasiticus strain

Rice husks (RHs) are plant waste materials abundant in phytoliths silica bodies. These were used as starting material for fungal-mediated biotransformation leading to the synthesis of a high-value added product. A strain of Aspergillus parasiticus was capable of transforming the amorphous silica conglomerates into structured nanoparticles (NPs) in the process of RHs biotransformation. Silica NPs were produced extracellularly and their size ranged from 3 to 400 nm depending on the biotransformation conditions and the post-biotransformation supernatant processing. To characterize the NP's structure and dimension, SEM, STEM, EDX and FTIR technics were applied. These demonstrated and confirmed that pyramid (400 nm), cubical (85 nm) and spherical (3 nm and 24 ± 8 nm) forms of silica NPs were obtained.     

Wernicke-Korsakoff syndrome as a rare phenotype of sporadic Creutzfeldt-Jakob disease

We reported the case of a patient with Wernicke-Korsakoff syndrome (WKs) as an early clinical manifestation of sporadic Creutzfeld-Jakob disease (sCJD). The 66-year-old female complained of dizziness and imbalance which mostly occurred while walking. A neurological examination revealed a triad of symptoms characteristic for WKs such as gaze paresis, ataxia of limbs and trunk as well as memory disturbances with confabulations. The disturbances increased during the course of the disease, which led to the death of the patient four months after the appearance of the signs. The patient was finally diagnosed with sCJD disease. The most useful ancillary examination results supporting sCJD diagnosis were brain diffusion DWI MRI (diffusion weighted magnetic resonance imaging) and the presence of 14–3-3 protein in CSF (cerebrospinal fluid). Since that manifestation of sCJD is very unique other causes should be taken into consideration while making a final diagnosis.     

Association mapping of drought tolerance-related traits in barley to complement a traditional biparental QTL mapping study

Key message

Association mapping of drought-related traits in barley was used to increase the density of existing QTL maps without recreating mapping populations.

Abstract

We used 109 spring barley genotypes exhibiting high or low drought tolerance to elucidate the associations between diversity array technology sequencing (DArTseq) and single nucleotide polymorphism (SNP) markers and various physiological parameters related to plant responses to drought conditions. The study was performed in controlled conditions (growth chambers), drought tolerance was phenotyped in the four-leaf seedlings. We identified 58 associations including 34 new markers (i.e., 16 DArTseq and 18 SNP markers). The results for three markers were consistent with the data obtained in an earlier traditional biparental QTL mapping study. The regions neighboring markers on linkage group 2H contained the highest number of significant marker–trait associations. Five markers related to the photosynthetic activity of photosystem II were detected on chromosome 4H. The lowest number of associations were observed for the sequences neighboring DArT markers on linkage group 6H. A chromosome 3H region related to water use efficiency and net photosynthesis rate in both biparental QTL, and association study, may be particularly valuable, as these parameters correspond to the ability of plants to remain highly productive under water deficit stress. Our findings confirm that association mapping can increase the density of existing QTL maps without recreating mapping populations.