The new unique technology for granulation of metals and nonmetallic melts is presented in the paper. It was created on the newly developed controlled film flow disintegration with rapid cooling of the droplets and solidified granules in amorphous state. Three new phenomena discovered by us created the base for 3 different methods for granulation of metals and nonmetallic melts in a wide range of granules’ sizes. Electromagnetic resonance method allows producing granules of metals in the range of 1 mm and over. The soliton-like vibrational regime is available for production of any granulation material in the range 0.1-1 mm. Moreover, the shockwave regime is applicable for producing particles from 10 micrometers by size. All methods produce particles of the stated size with a narrow distribution by size: deviation from the average size no more than 50%. The other known methods do not produce granules with such high cooling rate, which can realize the amorphous state and cannot regulate size of particles in such bright but narrow range from 10 micrometers to 1-2 mm.

Developing prolonged antigen delivery systems that mimick the long-term exposure to pathogens appears as a promising, still poorly explored approach to reach durable immunities. In this study, we have used a simple technology by which his-tagged proteins can be assembled, assisted by divalent cations, as supramolecular complexes with progressive complexity, namely protein-only nanoparticles and microparticles. Microparticles produced out of nanoparticles are biomimetics of secretory granules from the mammalian hormonal system. Upon subcutaneous administration, they slowly disintegrate, acting as an endocrine-like secretory system, and rendering the building block nanoparticles progressively bioavailable. The performance of such materials, previously validated for drug delivery in oncology, has been tested here regarding the potential for time-prolonged antigen release. This has been done by taking, as building blocks, a nanostructured version of p30, a main structural immunogen from the African swine fever virus (ASFV). By challenging the system in both mice and pigs, we have observed unusually potent pro-inflammatory activity in porcine macrophages, and long-lasting humoral and cellular responses in vivo, which might overcome the need for an adjuvant. The robustness of both innate and adaptive responses tag, for the first time, these dynamic depot materials as a novel and valuable instrument with transversal applicability in immune stimulation and vaccinology.

The tremendous diversity and complexity of proteins invariably results in protein misfolding, to which cells have evolved numerous mechanisms of mitigating. Degrading misfolded proteins is perhaps the most intuitive strategy, but also critical to managing proteostasis are the elaborate mechanisms of translational control. Attenuated rates of translation ameliorate protein misfolding by downregulating the flux of new protein and conserving ATP. Loss of translational control, particularly in neurons, constitutes a major proteostatic dysfunction capable of causing or exacerbating neurodegeneration, while interventions aimed at downregulating protein synthesis are generally neuroprotective. In this review, I examine the critical neuronal signaling networks employed to control translation with an emphasis on current research. This includes the Unfolded Protein Response (UPR), the mitochondrial UPR (mtUPR), mTORC1 signaling, and stress granule formation.

As the global demand for water increases, so does the quantity of wastewater requiring treatment. Due to a relatively low carbon footprint, compared with conventional wastewater treatment technologies, anaerobic digestion (AD) was identified in the 1970s as a forerunner in the push for sustainability, when interest in sustainable technologies and renewable energy sources was first sparked. AD technology development ultimately resulted in the discovery of the ‘anaerobic granule’. It is a spontaneously-forming bio-aggregate of microbial cells capable of digesting pollutants and producing methane-rich biogas as a renewable source of bioenergy. The high settling velocity of such granules meant that AD systems could be operated as high-rate treatment processes, because the active, relatively-slow-growing, pollutant-removing biomass would be retained inside, and not washed out of, even bioreactors operated at extremely high volumetric loading rates. In the intervening years the emergence of the anaerobic ammonium oxidising (anammox) granule, aerobic granule, hydrogenic granule, oxygenic photogranule, and many other functionally-specialised granules, has opened new opportunities in wastewater treatment biotechnology. Whilst environmental engineering based around wastewater treatment is still a growing field of research and implementation, the granule (in all forms) is starting to catch the attention of microbial ecologists. It is a self-immobilised biofilm, with many of the properties of ‘conventional’ biofilms formed in Nature. However, as a single entity, a granule represents an entire community of microorganisms, competing or functioning cooperatively or in syntrophy. Together, inside a bioreactor, granules perform side-by-side arguably representing a meta-organism. Granules are gaining traction as the perfect samples for high-throughput studies on fundamental ecological concepts. Granular biofilms can be used to test hypotheses around drivers of diversity, community assembly, biofilm formation and maturation, community expansion and succession, community stress response, among others. This review outlines the history of three of the most influential types of granules: the anaerobic (methanogenic), aerobic and anammox granule. The main biochemical processes found in each type; their primary characteristics; and the typical makeup of the microbial community underpinning the processes are compared. Finally, the adoption of granules as the perfect ‘playground’ for experiments in microbial ecology is reviewed.

A defining feature of a productive infection is the co-opting of host cell resources for viral replication. Despite their repertoire of molecular functions, viruses subvert host defenses to take control of cellular factors such as RNA binding proteins (RBPs). RBPs are involved in virtually all steps of mRNA life forming ribonucleoprotein complexes (mRNPs) in a highly ordered and regulated process to control RNA fate and stability in the cell. Thus, the hallmark of this viral takeover is to reshape RNA fate in the cell to modulate host gene expression and evade immune responses. Here we provide an extensive review of work in this area, particularly how the host-viral interplay influences RBP functions to modulate the host cell. Overall, in this review, we highlight the myriad of ways RBPs can regulate RNA stability in either a pro-viral or antiviral manner by gathering novel insights gained from research studies in this field.

Protein oligomerzation is key to countless physiological processes, but also to abnormal amyloid conformations implicated in over 25 mortal human diseases. Angiogenin (h-ANG), a ribonuclease A family member, produces RNA fragments that regulate ribosome formation, the creation of new blood vessels and stress granule function. Too little h-ANG activity leads abnormal protein oligomerization resulting in Amyotrophic Lateral Sclerosis (ALS) or Parkinson’s disease. While a score of disease linked h-ANG mutants has been studied by X-ray diffraction, some elude crystallization. There is also debate regarding the structure that RNA fragments adopt after cleavage by h-ANG. Here, to better understand the beginning of the process that leads to aberrant protein oligomerization, the solution secondary structure and residue-level dynamics of WT h-ANG and two mutants: H13A and R121C, are characterized by multidimensional heteronuclear NMR spectroscopy under near physiological conditions. All three variants are found to adopt well folded and highly rigid structures in solution, although the elements of secondary structure are somewhat shorter than those observed in cystallography studies. R121C alters the environment of nearby residues only. By constrast, the mutation H13A affects local residues as well as nearby active site residues residues K41 and H119. The conformation characterization by CD and 1D 1H NMR spectroscopies of tRNAAla before and after h-ANG cleavage reveals a retention of most duplex structure and little or no G-quadruplex formation.

Ralstonia solanacearum is the pathogen responsible for Ralstonia solanacearum, which is soil-borne, infests plant conduits to cause disease.They are not easy to control and are harmful.Therefore the development of a controlled-release agent responsive to the pathogen's soil temperature and moisture conditions could aid in its control. Corn kernel powder is used as a carrier for Tetramycin biopesticide, with various compounds tested for bonding.They were then made by extrusion moulding.Temperature and wetness responsive components, including Pentaerythritol Tetrastearate, Pentaerythritol Tetraoleate, Polyethylene Glycol Stearate, and Polyethylene Glycol Monooleate,are compounded and coated onto Tetramycin cores to form an intermediate shell layer. Finally, Ethyl cellulose(EC)and Hydroxypropyl methyl cellulose(HPMC) are dissolved and sprayed onto the outermost layer to complete the pellet's shell layer.After the evaluation of texture performance test,DSC analysis,TG test,temperature and humidity response performance test, as well as pellet cross-section SEM observation, field effectiveness test, etc., the preferred formulations were screened.The results showed that the preferred Core formulation:Corn kernel powder:Tetramycin aqueous:Xanthan gum mass ratio=13.5:23:2; Intermediate Shell layer:PETS: PETO: PEG400MO mass ratio=10:30:10; and Outer Shell layer: Ethyl cellulose (EC):Hydroxypropyl methyl cellulose (HPMC) mass ratio=5:1.Under soil conditions of high incidence of Ralstonia solanacearum, i.e., sustained warmth (30-35℃) and high humidity (30% moisture content), the formulated pills showed a release rate of 95.18% to 96.24% over a period of 35 days. Moreover, they showed a stepwise cyclic release process under alternating warm and humid-dry and cold conditions.In field tests on tobacco, a relative preventive efficacy of 54.74% was achieved 106 days post-transplanting. Soil bacterial community structure analysis revealed a significant decrease (2.28-2.84 times) in the abundance of Solanaceae Raelia under Tetramycin pellet treatment compared to the control. Additionally, fungal community structure showed highter consistency initially across various soil depths, with diversity differences between soil layers reestablished later (106 days), while the predominant categories of bacteria and fungi remained unchanged.In this study, we successfully developed a nucleoshell-type slow-controlled release pill embedded with Tetramycin. This pill effectively responded to temperature and humidity changes, resulting in high efficacy in disease prevention.This development holds significant promise for effectively improving drug utilization and easing application challenges, offering broad prospects for Ralstonia solanacearum prevention and control in agriculture.

The retinal pigment epithelium (RPE) is a monolayer of epithelial cells lying between the neural retina and the choroid and is essential for photoreceptor function and viability. Quail Coturnix japonica is a convenient experimental animal model for the study of age and pathological retina processes to an accelerated time regime. The three main types of pigment granules present in the RPE are melanin-containing melanosomes, lipofuscin-containing lipofuscin granules, and mixed melanolipofuscin granules containing both melanin and lipofuscin. It’s known that with aging and pathology in RPE cells the content of melanosomes decreases but lipofuscin and melanolipofuscin granules accumulate. We examined melanolipofuscin granules in macular areas of the quail retina by transmission electron microscopy in young, middle-aged, and old birds, and in middle-aged birds irradiated with blue LED light (450 nm, 4 J/cm2). It has been shown that during photooxidative stress caused by the action of blue light on the quail eye, active fusion of melanosomes and lipofuscin granules occurs with formation of various types, including giant, mixed melanolipofuscin-like granules. Irradiation result in almost twofold decrease in the number of lipofuscin granules and synchronous increase in the number of melanolipofuscin-like granules. Increased accumulation of melanolipofuscin-like granules was also observed in non-irradiated old birds. It is assumed that the decrease in the number of melanosomes in the RPE during aging and photo-oxidative stress is associated with their fusion with lipofuscin granules and subsequent degradation of melanin by reactive oxygen species formed in melanolipofuscin-like granules. The disappearance of melanin deprives the RPE cells of light-filtering and antioxidant protection, and significantly increases the risk of their oxidative stress.

It is known that during the process of aging there is a significant decrease in the number of melanosomes in the retinal pigment epithelium (RPE) cells of the human eye. We have previously shown that the interaction of melanin melanosomes with superoxide radicals results in its oxidative degradation with the formation of water-soluble fluorescent products. In the present study, we show using fluorescence analysis, HPLC, and mass spectrometry, that visible light irradiation of melanolipofuscin granules isolated from RPE cells of the human eye results in the formation of water-soluble fluorescent products of oxidative degradation of melanin, in contrast to lipofuscin granules and melanosomes irradiation. The formation of these products occurs as a result of oxidative degradation of melanin by superoxide radicals, which are generated by the lipofuscin part of the melanolipofuscin granule. We identified these products both in the composition of melanolipofuscin granules irradiated with visible light, and in composition of non-irradiated but oxidized by superoxide radicals, melanosomes. In melanolipofuscin granules irradiated with visible light, ions that could be associated to melanin oxidative degradation products were identified by applying principal component analysis of time-of-flight secondary ion mass spectrometry (ToF-SIMS) data. Degradation of the intact melanosomes by visible light is also possible; however, this requires significantly higher irradiation intensities than for melanolipofuscin granules. It is concluded that the decrease in the concentration of melanin in RPE cells of the human eye with age is due to its oxidative degradation by reactive oxygen species generated by lipofuscin as part of melanolipofuscin granules under the action of light.

Among the various methods for collecting oil spills and oil products, including from the water surface, one of the most effective is the use of sorbents. In this work, three-component bio-based composite granular adsorbents were produced and studied for oil products pollution collection. A bio-based binder made of peat, devulcanised crumb rubber from used tyres, and part fly ash as cenospheres were used for absorbent production. The structure, surface morphology, porosity, mechanical properties, and sorption kinetics of the obtained samples were studied. Composite hydrophobicity and sorption capacity to oil products such as diesel fuel (DF) and motor oil (MO) were determined. The obtained pellets are characterised by a sufficiently pronounced ability to absorb oil products such as DF. As the amount of CR in the granules increases, the diesel absorption capacity increases significantly. The case of 30-70-0 is almost 3 times higher than the granules from homogenised peat. The increase in q is due to two factors: pronounced surface hydrophobicity of the samples (Θ = 152°) and a heterogeneous porous granule structure. The presence of the cenosphere in the biocomposite reduces its surface hydrophobicity while increasing the diesel absorption capacity. Relatively rapid realisation of the maximum saturation by the MO was noted. In common, the designed absorbent shows up to 0.7 g·g^-1 sorption capacity for MO and up to 1.55 g·g^-1 sorption capacity for diesel. A possible mechanism of DF absorption and the limiting stages of the process approximated for different kinetic models are discussed. The Weber-Morris diffusion model is used to primarily distinguish the limiting effect of external and internal diffusion of adsorbate on the absorption process

Proteasomes critically regulate proteostasis via protein degradation. Proteasomes are multisubunit complexes composed of the 20S proteolytic core particle (20S CP) that, in association with one or two 19S regulatory particles (19S RPs), generate the 26S proteasome. 26S is the major proteasomal complex in the cells. Native gel protocols are used to investigate the 26S/20S ratio. However, a simple method for detecting these proteasome complexes in the cells is missing. To this end, using CRISPR technology, we YFP tagged the endogenous PSMB6 (b1) gene, a 20S CP subunit, and co-tagged endogenous PSMD6 (Rpn7), a 19S RP subunit, with mScarlet fluorescent protein. We observed colocalization of YFP and mScarlet fluorescent proteins in the cells, with higher nuclear accumulation. Nuclear proteasomal granules are formed under osmotic stress, and all were positive for YFP and mScarlet. Previously we have reported that PSMD1 knockdown, one of the 19 RP subunits, gives rise to a high level of “free” 20S CP. Intriguingly, under this condition, the 20S-YFP remained nuclear, whereas the PSMD6-mScarlet is mostly in cytoplasm, demonstrating the distinct subcellular distribution of uncapped 20S CP. Lately, we have shown that the PSMA3 (a7) C-termininus, a 20S CP subunit, binds multiple intrinsically disordered proteins (IDPs). Remarkably, truncation of the PSMA3 C-terminus is phenotypically reminiscent of PSMD1 knockdown. These data suggest the PSMA3 C-terminal region is critical for the 26S proteasome integrity.

Stress Granules formation is a pro-survival mechanism helping cells to cope with environmental challenges. Stress Granules have been studied for two decades in fundamental research, and are now being examined in the context of human pathogenesis. Here, we review studies highlighting stress granules’ involvement in cancer development through translational pattern modification.

Medullary thyroid carcinomas (MTCs) are rare thyroid tumors occurring in both sporadic and hereditary forms and whose pathogenesis is related to RET proto-oncogene alterations. MTCs originate from parafollicular cells, which produce calcitonin that represents the biochemical activity of MTC. Total thyroidectomy is the main treatment for MTC and often cures patients with confined diseases. In cases of metastasis, the approach depends on the rate of progression of disease. We report a case of a 54 years old female with a single, incidentally discovered, thyroid nodule of 1 cm, classified as suspicious MTC after a stimulation test with i.v. calcium. After surgery, we examined the nodule using immunohistochemistry, immunofluorescence and electron microscopy. In addition to calcitonin, we found that it expressed intracellular positivity for the RTK receptors ERBB1 and ERBB2. Consistently with MTC features, ultrastructural examination of the tumor displayed heterogeneous spindle-shaped cells containing two groups of secretory granules. Due to the significant correlation found between high ERBB1/ERBB2 levels in MTCs and extrathyroidal growth, the detection of ERBB1 and ERBB2 expression suggests that the two oncoproteins may possibly be involved in tumor proliferative responses and/or differentiation of C-cells. The biological, prognostic and therapeutic significance of these patterns would merits further investigations.

Autophagy was initially recognized as a bulk degradation process that randomly sequesters and degrades cytoplasmic material in lysosomes (vacuoles in yeast). In recent years, various selective autophagy pathways have been discovered. Glycophagy, the selective autophagy of glycogen granules, is one of them. It is an important contributor to Pompe disease, a type of lysosomal storage disorder with lysosomal accumulation of glycogen. Here, we developed the Komagataella phaffii yeast as a simple model of glycogen autophagy under nitrogen starvation conditions. For this, we turned the self-glucosylating initiator of glycogen synthesis, Glg1, which is covalently bound to glycogen, into the Glg1-GFP autophagic reporter. Our results revealed that vacuolar delivery of Glg1-GFP and its processing to free GFP were strictly dependent on autophagic machinery and vacuolar proteolysis. Notably, this process was independent of Atg11, the scaffold protein common for many selective autophagy pathways. Importantly, the non-mutated Glg1-GFP (which synthesizes and marks glycogen) and mutated Glg1Y212F-GFP (which does not synthesize glycogen and is degraded by non-selective autophagy as cytosolic Pgk1-GFP) were equally well delivered to the vacuole and had similar levels of released GFP. Therefore, we concluded that glycogen autophagy is a non-selective process in K. phaffii yeast under nitrogen starvation conditions.

Extracellular vesicles (EV) are a very heterogeneous group of cell-derived vesicles released by almost all kind of living cells. EV are involved in intercellular communication, both locally and systemically, since they induce signals and transfer their contents (proteins, lipids, RNAs) to other cells, which subsequently trigger a wide variety of biological responses in the target cells. How-ever, cell surface receptor-induced EV release is limited to cells from the immune system, includ-ing T lymphocytes. T cell receptor activation of T lymphocytes induces secretion of EV containing T cell receptor for antigen and several bioactive molecules, including proapoptotic proteins. These EV are thus specific for antigen-bearing cells, which make them ideal candidates for a cell-free, EV-dependent cancer therapy. In this review we discuss the generation of EV by T lymphocytes and some potential therapeutic approaches of these EV.

Cellular biomolecular condensates termed ribonucleoprotein (RNP) granules often are enriched in messenger RNA (mRNA) molecules relative to the surrounding cytoplasm. Yet, the spatial localization and diffusion of mRNAs in close proximity to phase separated RNP granules is not well understood. In this study, we performed single molecule fluorescence imaging experiments of mRNAs in live cells in the presence of two types of RNP granules, stress granules (SG) and processing bodies (PB), which are distinct in their molecular composition and function. We developed a photobleaching- and noise-corrected colocalization imaging algorithm that was employed to determine the accurate positions of individual mRNAs relative to the granule’s boundaries. We found that mRNAs are often localized at granule boundaries, an observation consistent with recently published data. We suggest that mRNA molecules become spontaneously confined at the RNP granule boundary similar to the adsorption of polymer molecules at liquid-liquid interfaces, which is observed in various technological and biological processes. We also suggest that this confinement could be due to a combination of intermolecular interactions associated with, first, the screening of a portion of the RNP granule interface by the polymer and, second, electrostatic interactions due to a strong electric field induced by a Donnan potential generated across the thin interface.

Since its initial characterization in 2016, the interferon stimulated gene Shiftless (SHFL) has proven to be a critical piece of the innate immune response to viral infection. SHFL expression stringently restricts the replication of multiple DNA, RNA, and retroviruses with an extraordinary diversity of mechanisms that differ from one virus to the next. These inhibitory strategies include the negative regulation of viral RNA stability, translation, and even the manipulation of RNA granule formation during viral infection. Even more surprisingly, SHFL is the first human protein found to directly inhibit the activity of the -1 programmed ribosomal frameshift, a translation recoding strategy utilized across nearly all domains of life and a several human viruses. Recent literature has shown that SHFL expression also significantly impacts viral pathogenesis in mouse models, highlighting its in-vivo efficacy. To help reconcile the many mechanisms by which SHFL restricts viral replication, we provide here a comprehensive review of this complex ISG, its influence over viral RNA fate, and the implications of its functions on the virus-host arms race for control of the cell.

Background: In regenerative dentistry the graft material influences the success. It should act as an osteoconductive scaffold, providing a mineral substrate during resorption and inducing the activity of osteoinductive cells capable of producing new bone, platelet growth factors, and cell differentiation factors inducing the differentiation of undifferentiated mesenchymal cells into. Given that dentin shares many biochemical characteristics with bone tissue, it has recently attracted considerable interest as a biomaterial for bone repair. The aim of this study is to compare two grinder types in order to determine the optimal method for producing dentinal particles using a mechanical grinder. Materials and methods: A sample of 40 natural human teeth without restorations, prostheses or root canal treatments was used and divided into two groups subjected to two different grinder speeds (high-speed and low-speed). The high-speed showed a greater dispersion (53.5+-9.89% of the tooth) due to the pulverisation (highly thin granules) of part of the tooth. The low-speed grinder does not pulverize the dentin and the percentage of tooth loss is 9.16+/-2.34%. Conclusion: The low-speed gringer allows to save a major part of the tooth and to have a maximum quantity of graft material but requires more time. Further studies must be promoted to optimise the grinding procedures.

At the molecular level, aging is often accompanied by dysfunction of stress-induced membrane-less organelles (MLOs) and changes in their material state. In this work, we analyzed the proteins included in the proteome of stress granules (SGs) and P-bodies for their tendency to transform the material state of these MLOs. Particular attention was paid to proteins whose gene expression changes during replicative aging. It was shown that the proteome of the studied MLOs practically does not differ in the analyzed characteristics and consists of completely or partially intrinsically disordered proteins, 30 - 40% of which are potentially capable of liquid-liquid phase separation (LLPS). At the same time, the proportion of proteins capable of spontaneous LLPS is relatively small, which indicates the leading role of nucleic acids in the biogenesis of these membrane-less organelles. Proteins whose gene expression changes during the transition of human cells to a senescent state make up about 20% of the studied proteomes. There is a statistically significant increase in the number of positively charged proteins in both datasets studied compared to the complete proteomes of these organelles. An increase in the relative content of DNA-, but not RNA-binding proteins, was also found in the stress-granules dataset with senescence-related processes. Among SG proteins potentially involved in senescent processes, there is an increase in the abundance of potentially amyloidogenic proteins compared to the whole proteome. The hnRPDL protein has the highest degree of disorder and highest propensity for LLPS among such proteins, which allows us to consider it as “potentially dangerous.” Proteins common to SGs and P bodies, potentially involved in processes associated with senescence, form clusters of interacting proteins. The largest cluster is represented by RNA-binding proteins involved in RNA processing and translation regulation. These data indicate that SG proteins, but not proteins of P-bodies, are more likely to transform the material state of MLOs. Furthermore, these MLOs can participate in processes associated with aging in a coordinated manner.