Karin Nilsson

RETRACTED: Vulnerability of Glioblastoma Cells to Catastrophic Vacuolization and Death Induced by a Small Molecule

Glioblastoma multiforme (GBM) is the most aggressive form of brain cancer with marginal life expectancy. Based on the assumption that GBM cells gain functions not necessarily involved in the cancerous process, patient-derived glioblastoma cells (GCs) were screened to identify cellular processes amenable for development of targeted treatments. The quinine-derivative NSC13316 reliably and selectively compromised viability. Synthetic chemical expansion reveals delicate structure-activity relationship and analogs with increased potency, termed Vacquinols. Vacquinols stimulate death by membrane ruffling, cell rounding, massive macropinocytic vacuole accumulation, ATP depletion, and cytoplasmic membrane rupture of GCs. The MAP kinase MKK4, identified by a shRNA screen, represents a critical signaling node. Vacquinol-1 displays excellent in vivo pharmacokinetics and brain exposure, attenuates disease progression, and prolongs survival in a GBM animal model. These results identify a vulnerability to massive vacuolization that can be targeted by small molecules and point to the possible exploitation of this process in the design of anticancer therapies.

The Molecular Mechanism of Peptide-mediated Erythromycin Resistance

The macrolide antibiotic erythromycin binds at the entrance of the nascent peptide exit tunnel of the large ribosomal subunit and blocks synthesis of peptides longer than between six and eight amino acids. Expression of a short open reading frame in 23 S rRNA encoding five amino acids confers resistance to erythromycin by a mechanism that depends strongly on both the sequence and the length of the peptide. In this work we have used a cell-free system for protein synthesis with components of high purity to clarify the molecular basis of the mechanism. We have found that the nascent resistance peptide interacts with erythromycin and destabilizes its interaction with 23 S rRNA. It is, however, in the termination step when the pentapeptide is removed from the peptidyl-tRNA by a class 1 release factor that erythromycin is ejected from the ribosome with high probability. Synthesis of a hexa- or heptapeptide with the same five N-terminal amino acids neither leads to ejection of erythromycin nor to drug resistance. We propose a structural model for the resistance mechanism, which is supported by docking studies. The rate constants obtained from our biochemical experiments are also used to predict the degree of erythromycin resistance conferred by varying levels of resistance peptide expression in living Escherichia coli cells subjected to varying concentrations of erythromycin. These model predictions are compared with experimental observations from growing bacterial cultures, and excellent agreement is found between theoretical prediction and experimental observation.

Electromagnetic forces in the air gap of a permanent magnet linear generator at no load

The basis for the work is the slow speed energy conversion of ocean wave energy into electricity using a direct-drive three-phase permanent magnetized linear generator. One of several important issues is the normal forces in the air gap, which is critical when designing the support structure of the generator. The electromagnetic forces in the air gap have been analyzed using Maxwell stress tensor method implemented in a two- dimensional finite element code. Simplified analytic calculations are made in order to validate the results from the extensive computer calculations. The normal electromagnetic forces in the air gap, Fδ, are analyzed for a two-sided linear generator at no load. An unstable condition of the global force on the piston occurs due to the fast increasing normal force as the air gap width decreases. A horizontal displacement of the piston from a neutral position with 3mm air gap on both sides produces a resulting horizontal force on the piston, increasing with the displacement. A displacement of 1mm gives a resulting horizontal force on the piston of 5.5kN per pole and meter of core length, which is increased to 9kN per pole and meter of core length for a displacement of 1.5mm. Furthermore, the normal force varies due to cogging as the piston moves vertically. At a constant air gap width of 3mm the normal forces per pole are varying between 9.9 and 11.3kN∕m of core length as the piston is moving from one pole to the next.The basis for the work is the slow speed energy conversion of ocean wave energy into electricity using a direct-drive three-phase permanent magnetized linear generator. One of several important issues is the normal forces in the air gap, which is critical when designing the support structure of the generator. The electromagnetic forces in the air gap have been analyzed using Maxwell stress tensor method implemented in a two- dimensional finite element code. Simplified analytic calculations are made in order to validate the results from the extensive computer calculations. The normal electromagnetic forces in the air gap, Fδ, are analyzed for a two-sided linear generator at no load. An unstable condition of the global force on the piston occurs due to the fast increasing normal force as the air gap width decreases. A horizontal displacement of the piston from a neutral position with 3mm air gap on both sides produces a resulting horizontal force on the piston, increasing with the displacement. A displaceme...

Erythromycin resistance by L4/L22 mutations and resistance masking by drug efflux pump deficiency

We characterized the effects of classical erythromycin resistance mutations in ribosomal proteins L4 and L22 of the large ribosomal subunit on the kinetics of erythromycin binding. Our data are consistent with a mechanism in which the macrolide erythromycin enters and exits the ribosome through the nascent peptide exit tunnel, and suggest that these mutations both impair passive transport through the tunnel and distort the erythromycin‐binding site. The growth‐inhibitory action of erythromycin was characterized for bacterial populations with wild‐type and L22‐mutated ribosomes in drug efflux pump deficient and proficient backgrounds. The L22 mutation conferred reduced erythromycin susceptibility in the drug efflux pump proficient, but not deficient, background. This ‘masking’ of drug resistance by pump deficiency was reproduced by modelling with input data from our biochemical experiments. We discuss the general principles behind the phenomenon of drug resistance ‘masking’, and highlight its potential importance for slowing down the evolution of drug resistance among pathogens.

Drug efflux pump deficiency and drug target resistance masking in growing bacteria

Recent experiments have shown that drug efflux pump deficiency not only increases the susceptibility of pathogens to antibiotics, but also seems to “mask” the effects of mutations, that decrease the affinities of drugs to their intracellular targets, on the growth rates of drug-exposed bacteria. That is, in the presence of drugs, the growth rates of drug-exposed WT and target mutated strains are the same in a drug efflux pump deficient background, but the mutants grow faster than WT in a drug efflux pump proficient background. Here, we explain the mechanism of target resistance masking and show that it occurs in response to drug efflux pump inhibition among pathogens with high-affinity drug binding targets, low cell-membrane drug-permeability and insignificant intracellular drug degradation. We demonstrate that target resistance masking is fundamentally linked to growth-bistability, i.e., the existence of 2 different steady state growth rates for one and the same drug concentration in the growth medium. We speculate that target resistance masking provides a hitherto unknown mechanism for slowing down the evolution of target resistance among pathogens.

Pleiotrophin promotes vascular abnormalization in gliomas and correlates with poor survival in patients with astrocytomas.

Secretion of pleiotrophin by aggressive gliomas triggers the formation of abnormal blood vessels by endothelial cells. Growing blood vessels in gliomas Aggressive gliomas are particularly lethal, in part, because of increased density of blood vessels and abnormal vasculature that enables tumor growth and damages the brain. Various secreted factors, including VEGF and pleiotrophin, act on endothelial cells to promote blood vessel formation. By analyzing patient data, Zhang et al. correlated increased pleiotrophin abundance to more aggressive grades of glioma and decreased survival. When implanted in mice, glioma cells that released pleiotrophin formed larger tumors with more blood vessels and increased VEGF concentrations near the blood vessels. Mice had smaller gliomas and survived longer when treated with inhibitors of ALK, a receptor for pleiotrophin, or an inhibitor of the VEGF receptor. These results suggest that blocking the signals that promote the abnormal blood vessel growth may be beneficial to patients with aggressive gliomas. Glioblastomas are aggressive astrocytomas characterized by endothelial cell proliferation and abnormal vasculature, which can cause brain edema and increase patient morbidity. We identified the heparin-binding cytokine pleiotrophin as a driver of vascular abnormalization in glioma. Pleiotrophin abundance was greater in high-grade human astrocytomas and correlated with poor survival. Anaplastic lymphoma kinase (ALK), which is a receptor that is activated by pleiotrophin, was present in mural cells associated with abnormal vessels. Orthotopically implanted gliomas formed from GL261 cells that were engineered to produce pleiotrophin showed increased microvessel density and enhanced tumor growth compared with gliomas formed from control GL261 cells. The survival of mice with pleiotrophin-producing gliomas was shorter than that of mice with gliomas that did not produce pleiotrophin. Vessels in pleiotrophin-producing gliomas were poorly perfused and abnormal, a phenotype that was associated with increased deposition of vascular endothelial growth factor (VEGF) in direct proximity to the vasculature. The growth of pleiotrophin-producing GL261 gliomas was inhibited by treatment with the ALK inhibitor crizotinib, the ALK inhibitor ceritinib, or the VEGF receptor inhibitor cediranib, whereas control GL261 tumors did not respond to either inhibitor. Our findings link pleiotrophin abundance in gliomas with survival in humans and mice, and show that pleiotrophin promotes glioma progression through increased VEGF deposition and vascular abnormalization.

Glioma-derived macrophage migration inhibitory factor (MIF) promotes mast cell recruitment in a STAT5-dependent manner.

Recently, glioma research has increased its focus on the diverse types of cells present in brain tumors. We observed previously that gliomas are associated with a profound accumulation of mast cells (MCs) and here we investigate the underlying mechanism.

Comparative drug pair screening across multiple glioblastoma cell lines reveals novel drug-drug interactions

BACKGROUND Glioblastoma multiforme (GBM) is the most aggressive brain tumor in adults, and despite state-of-the-art treatment, survival remains poor and novel therapeutics are sorely needed. The aim of the present study was to identify new synergistic drug pairs for GBM. In addition, we aimed to explore differences in drug-drug interactions across multiple GBM-derived cell cultures and predict such differences by use of transcriptional biomarkers. METHODS We performed a screen in which we quantified drug-drug interactions for 465 drug pairs in each of the 5 GBM cell lines U87MG, U343MG, U373MG, A172, and T98G. Selected interactions were further tested using isobole-based analysis and validated in 5 glioma-initiating cell cultures. Furthermore, drug interactions were predicted using microarray-based transcriptional profiling in combination with statistical modeling. RESULTS Of the 5 × 465 drug pairs, we could define a subset of drug pairs with strong interaction in both standard cell lines and glioma-initiating cell cultures. In particular, a subset of pairs involving the pharmaceutical compounds rimcazole, sertraline, pterostilbene, and gefitinib showed a strong interaction in a majority of the cell cultures tested. Statistical modeling of microarray and interaction data using sparse canonical correlation analysis revealed several predictive biomarkers, which we propose could be of importance in regulating drug pair responses. CONCLUSION We identify novel candidate drug pairs for GBM and suggest possibilities to prospectively use transcriptional biomarkers to predict drug interactions in individual cases.

Direct electric energy conversion system for energy conversion from marine currents

Abstract Direct electric energy conversion without input power regulating system or gearboxes may offer a simpler and thus low cost way of converting the energy of moving water. The proposed electrical system consists of a permanent magnetized direct drive generator, a rectification step, transmission, an inverter, a transformer, and a grid connection. This paper focuses on the generator, and simulations of a 5 kW permanent magnetized direct drive generator have been performed. Operation of the generator at both part load and overloads are calculated and presented. It is found that this direct drive permanent magnetized generator provides a relatively high efficiency for both part load and overload conditions.

Converting kinetic energy in small watercourses using direct drive generators

Tidal currents, ocean currents and unregulated watercourses are all large sources of energy that can be converted into electricity. Several technical and economical solutions within this research area have been demonstrated. In literature there exists two different turbine types, horizontal and vertical axis. The present paper focuses on the design of a permanent magnetized generator directly coupled to a vertical axis turbine. The proposed theoretical concept is adapted to data measurements regarding water current velocities and flow profiles from a Swedish watercourse. A high electromagnetic efficiency of 90% is obtained.Copyright