Synthesis of Extracellular L-lysine-α-oxidase along with Degrading Enzymes by Trichoderma cf. aureoviride Rifai VKM F-4268D: Role in Biocontrol and Systemic Plant Resistance.

When cultivating on wheat bran or deactivated fungal mycelium as a model of "natural growth", the ability of Trichoderma to synthesize extracellular L-lysine-α-oxidase (LysO) simultaneously with cell-wall-degrading enzymes (proteases, xylanase, glucanases, chitinases, etc.), responsible for mycoparasitism, was shown. LysO, in turn, causes the formation of H2O2 and pipecolic acid. These compounds are known to be signaling molecules and play an important role in the induction and development of systemic acquired resistance in plants. Antagonistic effects of LysO have been demonstrated against phytopathogenic fungi and Gram-positive or Gram-negative bacteria with dose-dependent cell death. The antimicrobial effect of LysO decreased in the presence of catalase. The generating intracellular ROS in the presence of LysO was also shown in both bacteria and fungi, which led to a decrease in viable cells. These results suggest that the antimicrobial activity of LysO is due to two factors: the formation of exogenous hydrogen peroxide as a product of the enzymatic oxidative deamination of L-lysine and the direct interaction of LysO with the cell wall of the micro-organisms. Thus, LysO on its own enhances the potential of the producer in the environment; namely, the enzyme complements the strategy of the fungus in biocontrol and indirectly participates in inducing SAR and regulating the relationship between pathogens and plants.

Properties of Organosilicon Elastomers Modified with Multilayer Carbon Nanotubes and Metallic (Cu or Ni) Microparticles.

The structural and electro-thermophysical characteristics of organosilicon elastomers modified with multilayer carbon nanotubes (MWCNTs) synthesized on Co-Mo/Al2O3-MgO and metallic (Cu or Ni) microparticles have been studied. The structures were analyzed with scanning electron microscopy (SEM), transmission electron microscopy (TEM), Raman spectroscopy, and energy-dispersive X-ray spectroscopy (EDX). The main focus of this study was the influence of metallic dispersed fillers on the resistance of a modified elastomer with Cu and Ni to the degradation of electrophysical parameters under the action of applied electrical voltage. The distribution of the temperature field on the surface of a modified polymer composite with metallic micro-dimensional structures has been recorded. The collected data demonstrate the possibility of controlling the degradation caused by electrical voltage. It has been found that repeated on/off turns of the elastomer with an MWCNTs on 50 and 100 cycles leads to a deterioration in the properties of the conductive elastomer from the available power of 1.1 kW/m2 (-40 °C) and, as a consequence, a decrease in the power to 0.3 kW/m2 (-40 °C) after 100 on/off cycles. At the same time, the Ni additive allows increasing the power by 1.4 kW/m2 (-40 °C) and reducing the intensity of the degradation of the conductive structures (after 100 on/off cycles up to 1.2 kW/m2 (-40 °C). When Ni is replaced by Cu, the power of the modified composite in the heating mode increases to 1.6 kW/m2 (-40 °C) and, at the same time, the degradation of the conductive structures in the composite decreases in the mode of cyclic offensives (50 and 100 cycles) (1.5 kW/m2 (-40 °C)). It was found that the best result in terms of heat removal is typical for an elastomer sample with an MWCNTs and Cu (temperature reaches 93.9 °C), which indicates an intensification of the heat removal from the most overheated places of the composite structure. At the same time, the maximum temperature for the Ni additives reaches 86.7 °C. A sample without the addition of a micro-sized metal is characterized by the local unevenness of the temperature field distribution, which causes undesirable internal overheating and destruction of the current-conducting structures based on the MWCNTs. The maximum temperature at the same time reaches a value of 49.8 °C. The conducted studies of the distribution of the micro-sizes of Ni and Cu show that Cu, due to its larger particles, improves internal heat exchange and intensifies heat release to the surface of the heater sample, which improves the temperature regime of the MWCNTs and, accordingly, increases resistance to electrophysical degradation.

Histone Abundance Quantification via Flow Cytometry of Htb2-GFP Allows Easy Monitoring of Cell Cycle Perturbations in Living Yeast Cells, Comparable to Standard DNA Staining.

Assaying changes in the amount of DNA in single cells is a well-established method for studying the effects of various perturbations on the cell cycle. A drawback of this method is the need for a fixation procedure that does not allow for in vivo study nor simultaneous monitoring of additional parameters such as fluorescence of tagged proteins or genetically encoded indicators. In this work, we report on a method of Histone Abundance Quantification (HAQ) of live yeast harboring a GFP-tagged histone, Htb2. We show that it provides data highly congruent with DNA levels, both in Saccharomyces cerevisiae and Ogataea polymorpha yeasts. The protocol for the DNA content assay was also optimized to be suitable for both Ogataea and Saccharomyces yeasts. Using the HAQ approach, we demonstrate the expected effects on the cell cycle progression for several compounds and conditions and show usability in conjunction with additional fluorophores. Thus, our data provide a simple approach that can be utilized in a wide range of studies where the effects of various stimuli on the cell cycle need to be monitored directly in living cells.

Comparative analysis of calcified soft tissues revealed shared deregulated pathways.

Introduction: Calcification of soft tissues is a common age-related pathology that primarily occurs within vascular tissue. The mechanisms underlying pathological calcification in humans and tissue specificity of the process is still poorly understood. Previous studies examined calcified tissues on one to one basis, thus preventing comparison of deregulated pathways across tissues. Purpose: This study aimed to establish common and tissue-specific changes associated with calcification in aorta, artery tibial, coronary artery and pituitary gland in subjects from the Genotype-Tissue Expression (GTEx) dataset using its RNA sequencing and histological data. Methods: We used publicly available data from the GTEx database https://gtexportal.org/home/aboutGTEx. All GTEx tissue samples were derived by the GTEx consorcium from deceased donors, with age from 20 to 79, both men and women. GTEx study authorization was obtained via next-of-kin consent for the collection and banking of de-identified tissue samples for scientific research. Hematoxylin and eosin (H&E) staining of arteries were manually graded based on the presence of calcification on a scale from zero to four, where zero designates absence of calcification and four designates severe calcification. Samples with fat contamination and mislabeled tissues were excluded, which left 430 aorta, 595 artery tibial, 124 coronary artery, and 283 pituitary samples for downstream gene expression analysis. Transcript levels of protein-coding genes were associated with calcification grade using sex, age bracket and cause of death as covariates, and tested for pathway enrichment using gene set enrichment analysis. Results: We identified calcification deposits in 28 (6.5%) aortas, 121 (20%), artery tibials, 54 (43%), coronary arteries, and 24 (8%) pituitary glands of GTEx subjects. We observed an age-dependent increase in incidence of calcification in all vascular tissues, but not in pituitary. Subjects with calcification in the artery tibial were significantly more likely to have calcification in the coronary artery (OR = 2.56, p = 6.3e-07). Markers of calcification previously established in preclinical and in vitro studies, e.g., BMP2 and RUNX2, were deregulated in the calcified tibial and coronary arteries, confirming the relevance of these genes to human pathology. Differentially expressed genes associated with calcification poorly overlapped across tissues suggesting tissue-specific nuances in mechanisms of calcification. Nevertheless, calcified arteries unanimously down-regulated pathways of intracellular transport and up-regulated inflammatory pathways suggesting these as universal targets for pathological calcification. In particular, PD-1 and PD-L1 genes were up-regulated in calcified tissues but not in the blood of the same subjects, suggesting that localized inflammation contributes to pathological calcification. Conclusion: Pathological calcification is a prevalent disease of aging that shares little changes in expression in individual genes across tissues. However, our analysis suggests that it potentially can be targeted by alleviating local inflammation of soft tissues.

Metal-Organic Framework (MOF)-A Universal Material for Biomedicine.

Metal-organic frameworks (MOFs) are a very promising platform for applications in various industries. In recent years, a variety of methods have been developed for the preparation and modification of MOFs, providing a wide range of materials for different applications in life science. Despite the wide range of different MOFs in terms of properties/sizes/chemical nature, they have not found wide application in biomedical practices at present. In this review, we look at the main methods for the preparation of MOFs that can ensure biomedical applications. In addition, we also review the available options for tuning the key parameters, such as size, morphology, and porosity, which are crucial for the use of MOFs in biomedical systems. This review also analyses possible applications for MOFs of different natures. Their high porosity allows the use of MOFs as universal carriers for different therapeutic molecules in the human body. The wide range of chemical species involved in the synthesis of MOFs makes it possible to enhance targeting and prolongation, as well as to create delivery systems that are sensitive to various factors. In addition, we also highlight how injectable, oral, and even ocular delivery systems based on MOFs can be used. The possibility of using MOFs as therapeutic agents and sensitizers in photodynamic, photothermal, and sonodynamic therapy was also reviewed. MOFs have demonstrated high selectivity in various diagnostic systems, making them promising for future applications. The present review aims to systematize the main ways of modifying MOFs, as well as the biomedical applications of various systems based on MOFs.

Changes in the Electrophysical Parameters of Nanomodified Elastomers Caused by Electric Current's Passage.

The development of reliable and effective functional materials that can be used in various technological fields and environmental conditions is one of the goals of modern nanotechnology. Heating elements' manufacturing requires understanding the laws of heat transfer under conditions of different supply voltages, as this expands the possibilities of such materials' application. Elastomers based on silicon-organic compounds and polyurethane modified with multi-walled carbon nanotubes (MWCNTs) were studied at various concentrations of Ni/MgO or Co-Mo/MgO and voltages (220, 250, and 300 V). It was found that an increase in voltage from 220 to 300 V leads to an initial increase in specific power on one-third followed by a subsequent decrease in a specific power when switched on again to 220 V (for -40 °C) of up to ~44%. In turn, for a polyurethane matrix, an increase in voltage to 300 V leads to an initial peak power value of ~15% and a decrease in power when switched on again by 220 V (for -40 °C) to ~36% (Ni/MgO -MWCNT). The conducted studies have shown that the use of a polyurethane matrix reduces power degradation (associated with voltage surges above 220 V) by 2.59% for Ni/MgO-based MWCNT and by 10.42% for Co-Mo/MgO. This is due to the better heat resistance of polyurethane and the structural features of the polymer and the MWCNT. The current studies allow us to take the next step in the development of functional materials for electric heating and demonstrate the safety of using heaters at a higher voltage of up to 300 V, which does not lead to their ignition, but only causes changes in electrophysical parameters.

Isolation of Clinical Microbial Isolates during Orthodontic Aligner Therapy and Their Ability to Form Biofilm.

The purpose of this study is to calculate microbiological composition of aligners after a day of wearing them. To date, the dental market for orthodontists offers many ways to correct bites. Aligners are transparent and almost invisible from the teeth. They are used for everyday wear to correct the incorrect position of the teeth, which was once considered the prerogative of braces. Scientists worldwide have repeatedly considered questions regarding the interaction between aligners and the oral cavity's microflora; however, the emphasis has mainly shifted toward species composition and antibiotic resistance. The various properties of these microorganisms, including biofilm formation, adhesion to various cells, and the ability to phagocytize, have not been studied so widely. In addition, these characteristics, as well as the microorganisms themselves, have properties that change over time, location, and in certain conditions. In this regard, the problem of biofilm formation in dental practice is always relevant. It requires constant monitoring since high contamination of orthodontic materials can reduce the effectiveness of local anti-inflammatory therapy and cause relapses in caries and inflammatory diseases of the oral cavity. Adhesive properties, one of the key factors in forming the architectonics of biofilms, provide the virulence factors of microorganisms and are characterized by an increase in optical density, determining the duration and retrospectivity of diagnostic studies. This paper focuses on the isolation of clinical microbial isolates during aligner therapy and their ability to form biofilms. In the future, we plan to use the obtained strains of microorganisms to create an effective and safe biofilm-destroying agent. We aimed to study morphometric and densitometric indicators of biofilms of microorganisms persisting on aligners.

Recent Advantages on Waste Management in Hydrogen Industry.

The turn to hydrogen as an energy source is a fundamentally important task facing the global energetics, aviation and automotive industries. This step would reduce the negative man-made impact on the environment on the one hand, and provide previously inaccessible power modes and increased resources for technical systems, predetermining the development of an absolutely new life cycle for important areas of technology, on the other. The most important aspect in this case is the development of next-generation technologies for hydrogen industry waste management that will definitely reduce the negative impact of technology on the environment. We consider the approaches and methods related to new technologies in the area of hydrogen storage (HS), which requires the use of specialized equipment equipped with efficient and controlled temperature control systems, as well as the involvement of innovative materials that allow HS in solid form. Technologies for controlling hydrogen production and storage systems are of great importance, and can be implemented using neural networks, making it possible to significantly improve all technological stages according to the criteria of energy efficiency reliability, safety, and eco-friendliness. The recent advantages in these directions are also reviewed.

Biopolymeric Nanoparticles-Multifunctional Materials of the Future.

Nanotechnology plays an important role in biological research, especially in the development of delivery systems with lower toxicity and greater efficiency. These include not only metallic nanoparticles, but also biopolymeric nanoparticles. Biopolymeric nanoparticles (BPNs) are mainly developed for their provision of several advantages, such as biocompatibility, biodegradability, and minimal toxicity, in addition to the general advantages of nanoparticles. Therefore, given that biopolymers are biodegradable, natural, and environmentally friendly, they have attracted great attention due to their multiple applications in biomedicine, such as drug delivery, antibacterial activity, etc. This review on biopolymeric nanoparticles highlights their various synthesis methods, such as the ionic gelation method, nanoprecipitation method, and microemulsion method. In addition, the review also covers the applications of biodegradable polymeric nanoparticles in different areas-especially in the pharmaceutical, biomedical, and agricultural domains. In conclusion, the present review highlights recent advances in the synthesis and applications of biopolymeric nanoparticles and presents both fundamental and applied aspects that can be used for further development in the field of biopolymeric nanoparticles.

Iodine Nanoparticles (Niodx™) for Radiotherapy Enhancement of Glioblastoma and Other Cancers: An NCI Nanotechnology Characterization Laboratory Study.

Effective and durable treatment of glioblastoma is an urgent unmet medical need. In this article, we summarize a novel approach of a physical method that enhances the effectiveness of radiotherapy. High atomic number nanoparticles that target brain tumors are intravenously administered. Upon irradiation, the nanoparticles absorb X-rays creating free radicals, increasing the tumor dose several fold. Radiotherapy of mice with orthotopic human gliomas and human triple negative breast cancers growing in the brain showed significant life extensions when the nanoparticles were included. An extensive study of the properties of the iodine-containing nanoparticle (Niodx) by the Nanotechnology Characterization Laboratory, including sterility, physicochemical characterization, in vitro cytotoxicity, in vivo immunological characterization, and in vivo toxicology, is presented. In summary, the iodine nanoparticle Niodx appears safe and effective for translational studies toward human use.

DNA microcircles - The promising tool for in vivo studies of the behavior of non-canonical DNA.

The paper discusses the reasons for the resurrection of the term DNA microcircles through the change of its definition to "topologically closed DNA circles with the length less than 1 Kbp" from the entire population of circular DNA that holds the name of minicircles. The possible applications of such tool for in vivo studies of non-canonical DNA are also discussed. Prospective for in vivo and in vitro studies of non-canonical DNA cloned into microcircles are demonstrated. A method of stepwise elongation or shortening of plasmids is discussed.

Novel Iodine nanoparticles target vascular mimicry in intracerebral triple negative human MDA-MB-231 breast tumors.

Triple negative breast cancer (TNBC), ~ 10-20% of diagnosed breast cancers, metastasizes to brain, lungs, liver. Iodine nanoparticle (INP) radioenhancers specifically localize to human TNBC MDA-MB-231 tumors growing in mouse brains after iv injection, significantly extending survival of mice after radiation therapy (RT). A prominent rim of INP contrast (MicroCT) previously seen in subcutaneous tumors but not intracerebral gliomas, provide calculated X-ray dose-enhancements up to > eightfold. Here, MDA-MB-231-cells, INPs, CD31 were examined by fluorescence confocal microscopy. Most INP staining co-localized with CD31 in the tumor center and periphery. Greatest INP/CD31 staining was in the tumor periphery, the region of increased MicroCT contrast. Tumor cells are seen to line irregularly-shaped spaces (ISS) with INP, CD31 staining very close to or on the tumor cell surface and PAS stain on their boundary and may represent a unique form of CD31-expressing vascular mimicry in intracerebral 231-tumors. INP/CD31 co-staining is also seen around ISS formed around tumor cells migrating on CD31+ blood-vessels. The significant radiation dose enhancement to the prolific collagen I containing, INP-binding ISS found throughout the tumor but concentrated in the tumor rim, may contribute significantly to the life extensions observed after INP-RT; VM could represent a new drug/NP, particularly INP, tumor-homing target.

Roadmap for metal nanoparticles in radiation therapy: current status, translational challenges, and future directions.

This roadmap outlines the potential roles of metallic nanoparticles (MNPs) in the field of radiation therapy. MNPs made up of a wide range of materials (from Titanium, Z = 22, to Bismuth, Z = 83) and a similarly wide spectrum of potential clinical applications, including diagnostic, therapeutic (radiation dose enhancers, hyperthermia inducers, drug delivery vehicles, vaccine adjuvants, photosensitizers, enhancers of immunotherapy) and theranostic (combining both diagnostic and therapeutic), are being fabricated and evaluated. This roadmap covers contributions from experts in these topics summarizing their view of the current status and challenges, as well as expected advancements in technology to address these challenges.

Iodine nanoparticles enhance radiotherapy of intracerebral human glioma in mice and increase efficacy of chemotherapy.

Gliomas and other brain tumors have evaded durable therapies, ultimately causing about 20% of all cancer deaths. Tumors are widespread in the brain at time of diagnosis, limiting surgery and radiotherapy effectiveness. Drugs are also poorly effective. Radiotherapy (RT) is limited by dose to normal tissue. However, high-atomic-number elements absorb X-rays and deposit the absorbed dose locally, even doubling (or more) the local dose. Previously we showed that gold nanoparticles (AuNPs) with RT could eradicate some brain tumors in mice and many other preclinical studies confirmed AuNPs as outstanding radioenhancers. However, impediments to clinical translation of AuNPs have been poor clearance, skin discoloration, and cost. We therefore developed iodine nanoparticles (INPs) that are almost colorless, non-toxic, lower cost, and have reasonable clearance, thus overcoming major drawbacks of AuNPs. Here we report the use of iodine nanoparticle radiotherapy (INRT) in treating advanced human gliomas (U87) grown orthotopically in nude mice resulting in a more than a doubling of median life extension compared to RT alone. Significantly, INRT also enhanced the efficacy of chemotherapy when it was combined with the chemotherapeutic agent Doxil, resulting in some longer-term survivors. While ongoing optimization studies should further improve INRT, clinical translation appears promising.

Small, Long Blood Half-Life Iodine Nanoparticle for Vascular and Tumor Imaging.

Standard clinical X-ray contrast agents are small iodine-containing molecules that are rapidly cleared by the kidneys and provide robust imaging for only a few seconds, thereby limiting more extensive vascular and tissue biodistribution imaging as well as optimal tumor uptake. They are also not generally useful for preclinical microCT imaging where longer scan times are required for high resolution image acquisition. We here describe a new iodine nanoparticle contrast agent that has a unique combination of properties: 20 nm hydrodynamic diameter, covalent PEG coating, 40 hour blood half-life, 50% liver clearance after six months, accumulation in tumors, and well-tolerated to at least 4 g iodine/kg body weight after intravenous administration in mice. These characteristics are unique among the other iodine nanoparticles that have been previously reported and provide extended-time high contrast vascular imaging and tumor loading. As such, it is useful for preclinical MicroCT animal studies. Potential human applications might include X-ray radiation dose enhancement for cancer therapy and vascular imaging for life-threatening situations where high levels of contrast are needed for extended periods of time.