PURPOSE: In carbon ion beam radiation therapy, fragmentation processes within the patient lead to changes in the composition of the particle field with increasing depth. Consequences are alterations of the resulting dose distribution and its biological effectiveness. To enable accurate treatment planning, the characteristics of the ion spectra resulting from fragmentation processes need to be known for various ion energies and target materials. In this work, we present a novel method for ion type identification using a small and highly flexible setup based on a single detector and designed to simplify measurements and overcome current shortages in available fragmentation data. MATERIALS AND METHODS: The presented approach is based on the pixelated, semiconductor detector Timepix. The large number of pixels with small pitch, all individually calibrated for energy deposition, enables detection and visualization of single particle tracks. For discrimination among different ion species, the pattern recognition analysis of the detector signal is used. Fragmentation spectra resulting from a primary carbon ion beam at various depths of tissue-equivalent material were studied to identify different ion species in mixed particle fields. The performance of the method was evaluated quantitatively using reference data from an established technique. RESULTS: All ion species resulting from carbon ion fragmentation in tissue-equivalent material could be separated. For measurements behind a 158-mm-thick water tank, the relative fractions of H, He, Be, and B ions detected agreed with corresponding reference data within the limits of uncertainty. For the relatively rare lithium ions, the agreement was within 2.3 Δref (uncertainty of reference). CONCLUSION: For designated configurations, the presented ion type identification method enables studies of therapeutic carbon ion beams with a simple, small, and configurable detection setup. The technique is promising to enable online fragmentation studies over a wide range of beam and target parameters in the future.

• Keywords
• carbon ion beam therapy, detector Timepix, ion type identification, nuclear fragmentation, pattern recognition analysis, pixel,
• Publication type
• Journal Article MeSH

Different explosive materials have been studied numerically and experimentally to assess the efficiency of a small diameter shaped charge in terms of produced jet characteristics and penetration depth into RHA steel targets. 26 different explosives have been simulated numerically using Autodyn hydrocode, whereas recommended explosives have been loaded into small diameter shaped charges by pressing technique and tested by static firing against RHA targets in order to validate the numerical calculations. The numerical analysis has presented an intensive global view about the variation of the shaped charge jets as a potential of the loaded explosive charge efficiencies. A successful trial has been performed to measure the shaped charge jet velocity using detonation velocity VOD 812 apparatus, where its measured value was only 3.6% different from the numerical one for HMX-V5 explosive. Besides, TITAN (L3) flash X-ray radiograph has also been implemented to explore the jet profile using the same explosive type and to measure its jet tip velocity, which has only 2.1% different from that estimated numerically. Extensive fragmentation analysis has been presented, which showed increase in both the fragment number and the fragment speed when the used explosive charge is of high detonation velocity. CL-20 explosive exhibited the largest jet tip velocity and its scaled collapse velocity was found to be 140% of TNT explosive. The calculated average fragment speed has been validated and the measured fragment speed has only 2.3% difference when compared to the SPH calculations.

• Keywords
• Autodyn, Flash X-ray radiograph, Fragmentation, HMX, Penetration, Plastic bonded explosives, Shaped charges,
• Publication type
• Journal Article MeSH

Multi-target drug discovery is one of the most followed approaches in the active central nervous system (CNS) therapeutic area, especially in the search for new drugs against Alzheimer's disease (AD). This is because innovative multi-target-directed ligands (MTDLs) could more adequately address the complexity of this pathological condition. In a continuation of our efforts aimed at a new series of anti-AD MTDLs, we combined the structural features of the cholinesterase inhibitor drug tacrine with that of resveratrol, which is known for its purported antioxidant and anti-neuroinflammatory activities. The most interesting hybrid compounds (5, 8, 9 and 12) inhibited human acetylcholinesterase at micromolar concentrations and effectively modulated Aβ self-aggregation in vitro. In addition, 12 showed intriguing anti-inflammatory and immuno-modulatory properties in neuronal and glial AD cell models. Importantly, the MTDL profile is accompanied by high-predicted blood-brain barrier permeability, and low cytotoxicity on primary neurons.

• Keywords
• Acetylcholinesterase, Alzheimer's disease, Amyloid, Multitarget compounds, Neuroinflammation,
• MeSH
• Acetylcholinesterase metabolism   MeSH
• Alzheimer Disease drug therapy   metabolism   MeSH
• Amyloid beta-Peptides chemistry   MeSH
• Antioxidants chemistry   metabolism   pharmacology   therapeutic use   MeSH
• Butyrylcholinesterase metabolism   MeSH
• Cholinesterase Inhibitors chemistry   metabolism   pharmacology   therapeutic use   MeSH
• Molecular Targeted Therapy * MeSH
• Blood-Brain Barrier metabolism   MeSH
• Liver drug effects   MeSH
• Rats MeSH
• Humans MeSH
• Ligands MeSH
• Neuroprotective Agents chemistry   pharmacology   therapeutic use   MeSH
• Peptide Fragments chemistry   MeSH
• Protein Aggregates drug effects   MeSH
• Drug Design * MeSH
• Resveratrol MeSH
• Stilbenes chemistry   MeSH
• Tacrine chemistry   metabolism   pharmacology   therapeutic use   MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Acetylcholinesterase MeSH
• amyloid beta-protein (1-42) MeSH Browser
• Amyloid beta-Peptides MeSH
• Antioxidants MeSH
• Butyrylcholinesterase MeSH
• Cholinesterase Inhibitors MeSH
• Ligands MeSH
• Neuroprotective Agents MeSH
• Peptide Fragments MeSH
• Protein Aggregates MeSH
• Resveratrol MeSH
• Stilbenes MeSH
• Tacrine MeSH

Thanks to the widespread use and safety profile of donepezil (1) in the treatment of Alzheimer's disease (AD), one of the most widely adopted multi-target-directed ligand (MTDL) design strategies is to modify its molecular structure by linking a second fragment carrying an additional AD-relevant biological property. Herein, supported by a proposed combination therapy of 1 and the quinone drug idebenone, we rationally designed novel 1-based MTDLs targeting Aβ and oxidative pathways. By exploiting a bioisosteric replacement of the indanone core of 1 with a 1,4-naphthoquinone, we ended up with a series of highly merged derivatives, in principle devoid of the "physicochemical challenge" typical of large hybrid-based MTDLs. A preliminary investigation of their multi-target profile identified 9, which showed a potent and selective butyrylcholinesterase inhibitory activity, together with antioxidant and antiaggregating properties. In addition, it displayed a promising drug-like profile.

• Keywords
• Alzheimer's disease, drug design, medicinal chemistry, multi-target drug discovery, polypharmacology,
• MeSH
• Acetylcholinesterase chemistry   metabolism   MeSH
• Alzheimer Disease drug therapy   MeSH
• Amyloid beta-Peptides antagonists & inhibitors   metabolism   MeSH
• Antioxidants chemistry   metabolism   pharmacology   MeSH
• Cholinesterase Inhibitors chemistry   metabolism   pharmacology   therapeutic use   MeSH
• Donepezil chemistry   metabolism   pharmacology   therapeutic use   MeSH
• Blood-Brain Barrier diagnostic imaging   metabolism   MeSH
• Indans chemistry   MeSH
• Humans MeSH
• Ligands * MeSH
• Cell Line, Tumor MeSH
• Neuroprotective Agents chemistry   metabolism   pharmacology   therapeutic use   MeSH
• Oxidative Stress drug effects   MeSH
• Protein Aggregates drug effects   MeSH
• Drug Design MeSH
• Cell Survival drug effects   MeSH
• Structure-Activity Relationship MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Acetylcholinesterase MeSH
• Amyloid beta-Peptides MeSH
• Antioxidants MeSH
• Cholinesterase Inhibitors MeSH
• Donepezil MeSH
• indacrinone MeSH Browser
• Indans MeSH
• Ligands * MeSH
• Neuroprotective Agents MeSH
• Protein Aggregates MeSH

Tamoxifen resistance remains a clinical obstacle in the treatment of hormone sensitive breast cancer. It has been reported that tamoxifen is able to target respiratory complex I within mitochondria. Therefore, we established two tamoxifen-resistant cell lines, MCF7 Tam5R and T47D Tam5R resistant to 5 μM tamoxifen and investigated whether tamoxifen-resistant cells exhibit mitochondrial changes which could help them survive the treatment. The function of mitochondria in this experimental model was evaluated in detail by studying i) the composition and activity of mitochondrial respiratory complexes; ii) respiration and glycolytic status; iii) mitochondrial distribution, dynamics and reactive oxygen species production. We show that Tam5R cells exhibit a significant decrease in mitochondrial respiration, low abundance of assembled mitochondrial respiratory supercomplexes, a more fragmented mitochondrial network connected with DRP1 Ser637 phosphorylation, higher glycolysis and sensitivity to 2-deoxyglucose. Tam5R cells also produce significantly higher levels of mitochondrial superoxide but at the same time increase their antioxidant defense (CAT, SOD2) through upregulation of SIRT3 and show phosphorylation of AMPK at Ser 485/491. Importantly, MCF7 ρ0 cells lacking functional mitochondria exhibit a markedly higher resistance to tamoxifen, supporting the role of mitochondria in tamoxifen resistance. We propose that reduced mitochondrial function and higher level of reactive oxygen species within mitochondria in concert with metabolic adaptations contribute to the phenotype of tamoxifen resistance.

• Keywords
• Breast cancer, Mitochondria, Mitochondrial fragmentation, Reactive oxygen species, Tamoxifen resistance,
• MeSH
• Apoptosis MeSH
• Cell Cycle MeSH
• Drug Resistance, Neoplasm * MeSH
• Phenotype MeSH
• Glycolysis * MeSH
• Antineoplastic Agents, Hormonal pharmacology   MeSH
• Humans MeSH
• Mitochondria metabolism   pathology   MeSH
• Mice, Nude MeSH
• Mice MeSH
• Tumor Cells, Cultured MeSH
• Breast Neoplasms drug therapy   metabolism   pathology   MeSH
• Cell Movement MeSH
• Cell Proliferation MeSH
• Reactive Oxygen Species metabolism   MeSH
• Electron Transport Complex I metabolism   MeSH
• Superoxides metabolism   MeSH
• Tamoxifen pharmacology   MeSH
• Xenograft Model Antitumor Assays MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Mice MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Antineoplastic Agents, Hormonal MeSH
• Reactive Oxygen Species MeSH
• Electron Transport Complex I MeSH
• Superoxides MeSH
• Tamoxifen MeSH

A BCL1 leukemia-cell-targeted polymer-drug conjugate with a narrow molecular weight distribution consisting of an N-(2-hydroxypropyl)methacrylamide copolymer carrier and the anticancer drug pirarubicin is prepared by controlled radical copolymerization followed by metal-free click chemistry. A targeting recombinant single chain antibody fragment (scFv) derived from a B1 monoclonal antibody is attached noncovalently to the polymer carrier via a coiled coil interaction between two complementary peptides. Two pairs of coiled coil forming peptides (abbreviated KEK/EKE and KSK/ESE) are used as linkers between the polymer-pirarubicin conjugate and the targeting protein. The targeted polymer conjugate with the coiled coil linker KSK/ESE exhibits 4× better cell binding activity and 2× higher cytotoxicity in vitro compared with the other conjugate. Treatment of mice with established BCL1 leukemia using the scFv-targeted polymer conjugate leads to a markedly prolonged survival time of the experimental animals compared with the treatment using the free drug and the nontargeted polymer-pirarubicin conjugate.

• Keywords
• cancer therapy, coiled coil, drug targeting, hydrophilic polymer, scFv,
• MeSH
• Acrylamides chemistry   MeSH
• Molecular Targeted Therapy MeSH
• Click Chemistry MeSH
• Cyclin D1 antagonists & inhibitors   immunology   MeSH
• Immunoglobulin Fragments administration & dosage   immunology   MeSH
• Immunoconjugates administration & dosage   chemistry   MeSH
• Drug Delivery Systems MeSH
• Leukemia immunology   pathology   therapy   MeSH
• Humans MeSH
• Disease Models, Animal MeSH
• Antibodies, Monoclonal chemistry   immunology   MeSH
• Mice MeSH
• Drug Carriers administration & dosage   chemistry   MeSH
• Peptides chemistry   immunology   MeSH
• Polymers administration & dosage   chemistry   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Acrylamides MeSH
• Cyclin D1 MeSH
• Immunoglobulin Fragments MeSH
• Immunoconjugates MeSH
• Antibodies, Monoclonal MeSH
• N-(2-hydroxypropyl)methacrylamide MeSH Browser
• Drug Carriers MeSH
• Peptides MeSH
• Polymers MeSH

Per- and polyfluoroalkyl substances (PFAS) are under regulatory scrutiny since some of them are persistent, bioaccumulative, and toxic. The occurrence of 4777 PFAS was investigated in the Danube River Basin (DRB; 11 countries) using target and suspect screening. Target screening involved investigation of PFAS with 56 commercially available reference standards. Suspect screening covered 4777 PFAS retrieved from the NORMAN Substance Database, including all individual PFAS lists submitted to the NORMAN Suspect List Exchange Database. Mass spectrometry fragmentation patterns and retention time index predictions of the studied PFAS were established for their screening by liquid chromatography - high resolution mass spectrometry using NORMAN Digital Sample Freezing Platform (DSFP). In total, 82 PFAS were detected in the studied 95 samples of river water, wastewater, groundwater, biota and sediments. Suspect screening detected 72 PFAS that were missed by target screening. Predicted no effect concentrations (PNECs) were derived for each PFAS via a quantitative structure-toxicity relationship (QSTR)-based approach and used for assessment of their environmental risk. Risk characterization revealed 18 PFAS of environmental concern in at least one matrix. The presence of PFAS in all studied environmental compartments across the DRB indicates a potentially large-scale migration of PFAS in Europe, which might require their further systematic regulatory monitoring.

• Keywords
• Danube River Basin, Ecotoxicological risk assessment, Environmental risk assessment, Per-and polyfluoroalkyl substances (PFAS), Suspect screening, Target screening,
• MeSH
• Water Pollutants, Chemical * analysis   MeSH
• Fluorocarbons * analysis   MeSH
• Wastewater analysis   MeSH
• Groundwater * chemistry   MeSH
• Rivers chemistry   MeSH
• Biota MeSH
• Water analysis   MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Water Pollutants, Chemical * MeSH
• Fluorocarbons * MeSH
• Waste Water MeSH
• Water MeSH

Liposomes functionalized with monoclonal antibodies or their antigen-binding fragments have attracted much attention as specific drug delivery devices for treatment of various diseases including cancer. The conjugation of antibodies to liposomes is usually achieved by covalent coupling using cross-linkers in a reaction that might adversely affect the characteristics of the final product. Here we present an alternative strategy for liposome functionalization: we created a recombinant Fab antibody fragment genetically fused on its C-terminus to the hydrophobic peptide derived from pulmonary surfactant protein D, which became inserted into the liposomal bilayer during liposomal preparation and anchored the Fab onto the liposome surface. The Fab-conjugated liposomes specifically recognized antigen-positive cells and efficiently delivered their cargo, the Alexa Fluor 647 dye, into target cells in vitro and in vivo. In conclusion, our approach offers the potential for straightforward development of nanomedicines functionalized with an antibody of choice without the need of harmful cross-linkers.

• Keywords
• Active targeting, Antibody engineering, Immunoliposome, Liposome functionalization, Recombinant Fab antibody fragment,
• MeSH
• CD48 Antigen metabolism   MeSH
• CD59 Antigens metabolism   MeSH
• Immunoglobulin Fab Fragments chemistry   immunology   metabolism   MeSH
• Jurkat Cells MeSH
• Humans MeSH
• Liposomes chemistry   MeSH
• Lymphoma immunology   metabolism   pathology   MeSH
• Antibodies, Monoclonal chemistry   immunology   metabolism   MeSH
• Mice MeSH
• Tumor Cells, Cultured MeSH
• Peptide Fragments immunology   metabolism   MeSH
• Pulmonary Surfactant-Associated Protein D immunology   metabolism   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• CD48 Antigen MeSH
• CD59 Antigens MeSH
• CD48 protein, human MeSH Browser
• CD59 protein, human MeSH Browser
• Immunoglobulin Fab Fragments MeSH
• Liposomes MeSH
• Antibodies, Monoclonal MeSH
• Peptide Fragments MeSH
• Pulmonary Surfactant-Associated Protein D MeSH

Objective.In the present hadrontherapy scenario, there is a growing interest in exploring the capabilities of different ion species other than protons and carbons. The possibility of using different ions paves the way for new radiotherapy approaches, such as the multi-ions treatment, where radiation could vary according to target volume, shape, depth and histologic characteristics of the tumor. For these reasons, in this paper, the study and understanding of biological-relevant quantities was extended for the case of4He ion.Approach.Geant4 Monte Carlo based algorithms for dose- and track-averaged LET (Linear Energy Transfer) calculations, were validated for4He ions and for the case of a mixed field characterised by the presence of secondary ions from both target and projectile fragmentation. The simulated dose and track averaged LETs were compared with the corresponding dose and frequency mean values of the lineal energy,yD¯andy¯F, derived from experimental microdosimetric spectra. Two microdosimetric experimental campaigns were carried out at the Italian eye proton therapy facility of the Laboratori Nazionali del Sud of Istituto Nazionale di Fisica Nucleare (INFN-LNS, Catania, I) using two different microdosimeters: the MicroPlus probe and the nano-TEPC (Tissue Equivalent Proportional Counter).Main results.A good agreement ofL¯dTotalandL¯tTotalwithy¯Dandy¯Texperimentally measured with both microdosimetric detectors MicroPlus and nano-TEPC in two configurations: full energy and modulated4He ion beam, was found.Significance.The results of this study certify the use of a very effective tool for the precise calculation of LET, given by a Monte Carlo approach which has the advantage of allowing detailed simulation and tracking of nuclear interactions, even in complex clinical scenarios.

• Keywords
• Geant4, Monte Carlo, linear energy transfer, microdosimetry, target fragmentation,
• MeSH
• Algorithms MeSH
• Ions MeSH
• Linear Energy Transfer * MeSH
• Monte Carlo Method MeSH
• Protons MeSH
• Radiometry * methods   MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Ions MeSH
• Protons MeSH

Cell death-inducing DFF[DNA fragmentation factor]-like effector-a (CIDEa), may initiate apoptosis by disrupting a complex consisting of 40-kDa caspase-3-activated nuclease (DFF40/CAD) and its 45-kDa inhibitor (DFF45/ICAD). CIDEa, however, was found to be localized in mitochondria. We have performed immunodetection of CIDEa in whole cells and subcellular fractions of HeLa cells adapted for a tetracycline-inducible CIDEa expression. Using immunocytochemistry we observed redistribution, enhanced upon treatment with camptothecin or valinomycin, of CIDEa to nucleus. Similarly, CIDEa content increased in the nuclear fraction but decreased in cytosolic fraction in cells treated to initiate apoptosis. We hypothesize that CIDEa is sequestered in mitochondria while transfer of this potentially dangerous protein from mitochondria into nucleus intensifies or even initiates apoptosis.

• MeSH
• Apoptosis MeSH
• Cell Nucleus metabolism   MeSH
• HeLa Cells MeSH
• Humans MeSH
• Mitochondria metabolism   MeSH
• Apoptosis Regulatory Proteins biosynthesis   metabolism   MeSH
• Protein Transport MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• CIDEA protein, human MeSH Browser
• Apoptosis Regulatory Proteins MeSH