Sugar decorated star-shaped (co)polymers with resveratrol-based core – physicochemical and biological properties

Star-shaped glycopolymers due to the attractive combination of the physicochemical, morphological, self-assembly properties along with biological activity have gained increased attention as innovative agents in novel cancer therapies. Unfortunately, the production of these highly desirable biomaterials remains a challenge in modern macromolecular chemistry. The main reason for that is the low polymerizability of ionic glycomonomers originated from their steric congestion and the occurrence of ionic interactions that generally negatively influence the polymerization progress and hinder controllable reaction pathway. In this work, the new ionic sugar monomer was (co)polymerized for the first time via Activator Generated by Electron Transfer Atom Transfer Radical Polymerization (AGET ATRP) using a three-arm resveratrol-based core to obtain star-like (co)polymers. The obtained products were examined in terms of their physicochemical properties and morphology. Aside from the synthesis of these new glycopolymers, also a thorough description of their thermal properties, ability to self-assembly, the formation of stable superstructures was studied in detail. It was found that examined (co)polymers did not show any heterogeneities and phase separation, while their variation of glass transition temperature (Tg) was strictly related to the change in the number of glycomonomer. Also, the stability and shapes of formed superstructures strictly depend on their composition and topology. Finally, we have shown that synthesized carbohydrate-based polymers revealed high antiproliferative activity against several cancer cell lines (i.e., breast, colon, glioma, and lung cancer). The cytotoxic activity was particularly observed for star-shaped polymers that were systematically enhanced with the growing concentration of amine moieties and molecular weight. The results presented herein suggest that synthesized star-shaped glyco(co)polymers are promising as drug or gene carriers in anticancer therapies or anti-tumor agents, depending on their cytotoxicity. Graphical abstract

Fabrication and Biological Activities of Plasmid DNA Gene Carrier Nanoparticles Based on Biodegradable l-Tyrosine Polyurethane

Gene therapy is a suitable alternative to chemotherapy due to the complications of drug resistance and toxicity of drugs, and is also known to reduce the occurrence of cellular mutation through the use of gene carriers. In this study, gene carrier nanoparticles with minimal toxicity and high transfection efficiency were fabricated from a biocompatible and biodegradable polymer, l-tyrosine polyurethane (LTU), which was polymerized from presynthesized desaminotyrosyl tyrosine hexyl ester (DTH) and polyethylene glycol (PEG), by using double emulsion and solvent evaporation techniques, resulting in the formation of porous nanoparticles, and then used to evaluate their potential biological activities through molecular controlled release and transfection studies. To assess cellular uptake and transfection efficiency, two model drugs, fluorescently labeled bovine serum albumin (FITC-BSA) and plasmid DNA-linear polyethylenimine (LPEI) complex, were successfully encapsulated in nanoparticles, and their transfection properties and cytotoxicities were evaluated in LX2 as a normal cell and in HepG2 and MCF7 as cancer cells. The morphology and average diameter of the LTU nanoparticles were confirmed using light microscopy, transmission electron microscopy, and dynamic light scattering, while confocal microscopy was used to validate the cellular uptake of FITC-BSA-encapsulated LTU nanoparticles. Moreover, the successful cellular uptake of LTU nanoparticles encapsulated with pDNA-LPEI and the high transfection efficiency, confirmed by gel electrophoresis and X-gal assay transfection, indicated that LTU nanoparticles had excellent cell adsorption ability, facilitated gene encapsulation, and showed the sustained release tendency of genes through transfection experiments, with an optimal concentration ratio of pDNA and LPEI of 1:10. All the above characteristics are ideal for gene carriers designed to transport and release drugs into the cytoplasm, thus facilitating effective gene therapy.

Active Surveillance in RET Gene Carriers Belonging to Families with Multiple Endocrine Neoplasia

Multiple Endocrine Neoplasia 2 (MEN2) is a hereditary cancer syndrome for developing medullary thyroid cancer (MTC) due to germline mutations of RET gene. Subjects harboring a germline RET mutation without any clinical signs of MTC are defined as gene carriers (GCs), for whom guidelines propose a prophylactic thyroid surgery. We evaluate if active surveillance of GCs, pursuing early thyroid surgery, can be safely proposed and if it allows safely delaying thyroid surgery in children until adolescence/adulthood. We prospectively followed 189 GCs with moderate or high risk germline RET mutation. Surgery was planned in case of: elevated basal calcitonin (bCT) and/or stimulated CT (sCT); surgery preference of subjects (or parents, if subject less than 18 years old); other reasons for thyroid surgery. Accordingly, at RET screening, we sub-grouped GCs in subjects who promptly were submitted to thyroid surgery (Group A, n = 67) and who were not (Group B, n = 122). Group B was further sub-grouped in subjects who were submitted to surgery during their active surveillance (Group B1, n = 22) and who are still in follow-up (Group B2, n = 100). Group A subjects presented significantly more advanced age, bCT and sCT compared to Group B. Mutation RETV804M was the most common variant in both groups but it was significantly less frequent in Group A than B. Analyzing age, bCT, sCT and genetic landscape, Group B1 subjects differed from Group B2 only for sCT at last evaluation. Group A subjects presented more frequently MTC foci than Group B1. Moreover, Group A MTCs presented more aggressive features (size, T and N) than Group B1. Accordingly, at the end of follow-up, all Group B1 subjects presented clinical remission, while 6 and 12 Group A MTC patients had structural and biochemical persistent disease, respectively. Thank to active surveillance, only 13/63 subjects younger than 18 years at RET screening have been operated on during childhood and/or adolescence. In Group B1, three patients, while actively surveilled, had the possibility to reach the age of 18 (or older) and two patients the age of 15, before being submitted to thyroid surgery. In Group B2, 12 patients become older than 18 years and 17 older than 15 years. In conclusion, we demonstrated that an active surveillance pursuing an early thyroid surgery could be safely recommended in GCs. This patient-centered approach permits postponing thyroid surgery in children until their adolescence/adulthood. At the same time, we confirmed that genetic screening allows finding hidden MTC cases that otherwise would be diagnosed much later.

Metacognitive insight into cognitive performance in pre and early-stage Huntington's disease

ABSTRACT Objectives Insight is an important predictor of quality of life in Huntington's disease and other neurodegenerative conditions. However, estimating insight with traditional methods such as questionnaires is challenging and subject to limitations. This study experimentally quantified metacognitive insight into cognitive performance in Huntington's disease gene-carriers. Methods We dissociated perceptual decision-making performance and metacognitive insight into performance in healthy controls (n=29), premanifest (n=19) and early-manifest (n=10) Huntington's disease gene-carriers. Insight was operationalised as the degree to which a participant's confidence in their performance was informative of their actual performance (metacognitive efficiency) and estimated using a computational model (HMeta-d). Results We found that pre and early-manifest Huntington's disease gene-carriers were impaired in making perceptual decisions compared to controls. Gene-carriers required more evidence in favour of the correct choice to achieve similar performance and perceptual impairments were increased in those with manifest disease. Surprisingly, despite marked perceptual impairments, Huntington's disease gene-carriers retained metacognitive insight into their perceptual performance. This was the case after controlling for confounding variables and regardless of disease stage. Conclusion We report for the first time a dissociation between impaired cognition and intact metacognition (trial-by-trial insight) in the early-stages of a neurodegenerative disease. This unexpected finding contrasts with the prevailing assumption that cognitive deficits are associated with impaired insight. Future studies should investigate how intact metacognitive insight could be used by some early Huntington's disease gene-carriers to positively impact their quality of life. Key words: Huntington's disease, decision-making, cognition, insight, metacognition

Estimating the Two Graph Dextran-Stearic Acid-Spermine Polymers Based on Iron Oxide Nanoparticles as Carrier for Gene Delivery

BackgroundNon-viral gene carriers because of their limited side effects, biocompatibility, simplicity and taking the advantages of electrostatic interactions have shown noticeable potential in gene delivery. The low transfection rate of non-viral vectors under physiological conditions is a significant issue. MethodsWe investigated the efficacy of hydrophilic and hydrophobic groups on gene carriers such as two synthesized amphiphilic polymer of dextran-stearic acid-spermine (DSASP) with verified lipid and amine conjugations that associated with Fe3O4 superparamagnetic nanoparticles to promote the target delivery and decrease the transfection time using static magnetic field.ResultsOur findings illustrate that magnetic nanoparticles are spherical with positive surface charges and superparamagnetic behaviors. The DSASP–pDNA/MNPs offered a strong pDNA condensation, protection against DNase degradation, significant cell viability in HEK 293T cells. Although conjugations of spermine play a critical role in transfection efficiency, amphiphilic polymer with more derivatives of stearic acid showed better transfection yields. ConclusionDSASP amphiphilic magnetic carriers offer new insights for gene delivery due to the amine contents and ameliorate the uptake of complexes via cell membrane based on its hydrophilic surface.

Marker features of the tomato basic material in heterosis breeding for greenhouse industry

Рассмотрены важные аспекты использования геноносителей различных маркерных признаков при создании и селекции гетерозисных гибридов томата для защищенного грунта. Наряду с общими требованиями к родительским компонентам, касающимися продуктивности растений, скороспелости, устойчивости к болезням и т. д., сегодня при селекции на гетерозис для защищенного грунта к исходному материалу предъявляют более высокие требования по наличию хозяйственно полезных признаков. Эти требования касаются типа роста растений, формы, окраски и качества плодов, типа кисти, формы плодоножки и т. д. Представлены характеристики исходного материала для создания гетерозисных гибридов томата, а также даны характеристики полученных гибридов. Исследования, проведенные в отделе селекции ВНИИО (ВНИИ овощеводства – филиал ФГБНУ ФНЦО), показали большую перспективность использования при гетерозисной селекции томата для защищенного грунта геноносителей различных маркерных признаков, особенно признаков, определяющих коричневую окраску эндосперма семян (гены bs и bs-2) и несочлененную плодоножку (ген j-2). Созданные с использованием этих маркерных признаков гибриды и селекционные линии по-своему уникальны и являются приоритетным достижением этого научного учреждения, так как они представляют большой интерес для гетерозисной селекции томата в качестве исходного материала. Целый ряд гибридов F1 включен в Госреестр селекционных достижений, допущенных к использованию (Удача, Голубчик, Клад овощевода, Заур и др.). В связи с развитием фермерского и любительского овощеводства в последние годы определенное внимание было уделено созданию новых мутантных линий с розовыми, малиновыми и темно-коричневыми плодами с высокими вкусовыми качествами (учитывая, что образцы такого типа пользуются повышенным спросом у населения и стоят дороже). При создании новых гетерозисных гибридов были учтены эти признаки родительских форм. Important aspects of the use of gene carriers of various marker traits in the creation and selection of heterotic tomato hybrids for greenhouse industry are considered. Along with the general requirements for the parent components concerning plant productivity, precocity, resistance to diseases, etc., today, when breeding for heterosis for greenhouse industry, higher requirements are imposed on the source material for the presence of economically useful features. These requirements relate to the type of plant growth, shape, colour and quality of fruits, brush type, stem shape, etc. The characteristics of the initial material for creating heterotic tomato hybrids are presented, as well as the characteristics of the resulting hybrids are given. Studies conducted in the breeding department of the ARRIVG (ARRIVG – a branch of the FSBI FSCV) showed great prospects for the use of gene carriers of various marker traits, especially those that determine the brown colour of the seed endosperm (bs and bs-2 genes) and an undifferentiated peduncle (j-2 gene) in the heterosis breeding of tomatoes for greenhouses. Hybrids and breeding lines created using these marker traits are unique in their own way and are a priority achievement of this scientific institution, since they are of great interest for the heterosis breeding of tomato as a basic material. A number of F1 hybrids are included in the State Register of Breeding Achievements allowed for use (Udacha, Golubchik, Klad ovoschevoda, Zaur, etc.). Due to the development of farming and amateur vegetable growing in recent years, some attention has been paid to the creation of new mutant lines with pink, crimson and dark brown fruits with high taste qualities (given that samples of this type are in high demand among the population and are more expensive). When creating new heterotic hybrids, these features of the parent forms were taken into account.