Preparation Of Suspensions Research Articles

Metal Ion-induced Gelation of High-concentration Graphite-like Crystalline Nanosheet Aqueous Suspensions.

The stability and transformation of nanomaterial aqueous suspensions are essential for their applications. Preparation of high-concentration carbon nanomaterials suspensions remains challenging due to their nonpolar nature. Herein, 200mgmL-1 carbon nanomaterial aqueous suspensions are achieved by using graphite-like crystalline nanosheets (GCNs) with high hydrophilicity. Furthermore, these high-concentration GCN aqueous suspensions spontaneously transform into gels when induced by mono-, di-, and trivalent metal salt electrolytes at room temperature. Theoretical calculation of potential energy by DLVO theory reveals that the gelatinized GCNs is a new and metastable state between two usual forms of solution and coagulation. It is shown that the gelation of GCNs is due to the preferential orientation of nanosheets in an edge-edge arrangement, which differs from the case of solution and coagulation. High-temperature treatment of GCN gels produces metal/carbon materials with pore structures. This work provides a promising opportunity to create various metal/carbon functional materials.

A Review of Freeze Casting: Preparation Process, Modified Methods, and Development Tendency

Abstract: Fabricating materials with nacre-like structure have received considerable attention as it shows an excellent combination of mechanical strength and toughness. A considerable number of researchers have reported the preparation method of bionic structure, such as layer-by-layer assembly, vacuum filtration, coextrusion assembly, electrophoresis deposition, water-evaporation-induced assembly, 3D printing, and freeze casting. Compared with other techniques, freeze casting, known as ice templating, is an environmentally friendly, prolongable, and potential method, so it has been rapidly developing and widely researched in recent decades. In this review, the front six methods with their benefits and limitations are briefly introduced. Then, the freeze casting technique with the preparation process and modified technique is emphatically analyzed. Finally, the future tendencies of materials application and technique application are discussed. Freeze casting consists of suspension preparation, solidification, sublimation, and post-treatment processes. The mechanism and influence of parameters during suspension preparation and solidification processes are principally discussed. It must be pointed out that the performance and structure of samples are closely related to the model and external force. Besides, the adjustable process parameters of freezing casting are a strong guarantee of obtaining the target product. The purpose of this review is to promote freeze casting workers to understand the influence of parameters and enlighten them in new experimental designs.

Uji Antidiare Ekstrak Etanol Daun Matoa (Pometia pinnata) Terhadap Mencit Jantan Diinduksi Oleum Ricini

Matoa leaf is a plant that is used as traditional medicine. Matoa leaves contain tannins, alkaloids, flavonoids, steroids, and glycosideswhich acts as an antidiarrheal. Oleum riciniis a triglyceride that is efficacious as a laxative. The purpose of this study was to prove that matoa leaves can be efficacious as an antidiarrheal.
 This study used an experimental method, using matoa leaf assay (Pometia pinnata J. R Forst & G. Forst.) with oleum ricini induction. This research includes the collection, processing, phytochemical screening, preparation of suspension of ethanol extract of matoa leaves, and testing the effectiveness antidiarrheal of suspension of ethanolic extract of matoa leaves (Pometia pinnata J.R. Forst& G. Forst.). The results of phytochemical screening on matoa leaf simplicia powder (Pometia pinnata J. R. Forst& G. Forst.) and ethanol extract of matoa leaf (Pometia pinnata J. R. Forst & G. Forst.) which has tannins, alkaloids, saponins, flavonoids, and triterpenoids. The results showed that EEDM at a dose of 300 mg/kg bw had an antidiarrheal effect comparable to EEDM at a dose of 150 mg/kg bw and had a stronger antidiarrheal effect than EEDM 75 mg/kg bw. The results also showed that the EEDM dose of 300 mg/kgbw was not significantly different from that of Loperamide HCl.

Atomic Force Microscopy Imaging of Elastin Nanofibers Self-Assembly.

Elastin is an extracellular matrix protein, providing elasticity to the organs, such as skin, blood vessels, lungs and elastic ligaments, presenting self-assembling ability to form elastic fibers. The elastin protein, as a component of elastin fibers, is one of the major proteins found in connective tissue and is responsible for the elasticity of tissues. It provides resilience to the human body, assembled as a continuous mesh of fibers that require to be deformed repetitively and reversibly. Thus, it is of great importance to investigate the development of the nanostructural surface of elastin-based biomaterials. The purpose of this research was to image the self-assembling process of elastin fiber structure under different experimental parameters such as suspension medium, elastin concentration, temperature of stock suspension and time interval after the preparation of the stock suspension. atomic force microscopy (AFM) was applied in order to investigate how different experimental parameters affected fiber development and morphology. The results demonstrated that through altering a number of experimental parameters, it was possible to affect the self-assembly procedure of elastin fibers from nanofibers and the formation of elastin nanostructured mesh consisting of naturally occurring fibers. Further clarification of the contribution of different parameters on fibril formation will enable the design and control of elastin-based nanobiomaterials with predetermined characteristics.

Comprehensive Review on Fabricating Bioactive Ceramic Bone Scaffold Using Vat Photopolymerization.

In recent years, bioactive ceramic bone scaffolds have drawn remarkable attention as an alternative method for treating and repairing bone defects. Vat photopolymerization (VP) is a promising additive manufacturing (AM) technique that enables the efficient and accurate fabrication of bioactive ceramic bone scaffolds. This review systematically reviews the research progress of VP-printed bioactive ceramic bone scaffolds. First, a summary and comparison of commonly used bioactive ceramics and different VP techniques are provided. This is followed by a detailed introduction to the preparation of ceramic suspensions and optimization of printing and heat treatment processes. The mechanical strength and biological performance of the VP-printed bioactive ceramic scaffolds are then discussed. Finally, current challenges and future research directions in this field are highlighted.

Nanomotion technology in combination with machine learning: a new approach for a rapid antibiotic susceptibility test for Mycobacterium tuberculosis

Nanomotion technology is a growth-independent approach that can be used to detect and record the vibrations of bacteria attached to cantilevers. We have developed a nanomotion-based antibiotic susceptibility test (AST) protocol for Mycobacterium tuberculosis (MTB). The protocol was used to predict strain phenotype towards isoniazid (INH) and rifampicin (RIF) using a leave-one-out cross-validation (LOOCV) and machine learning techniques. This MTB-nanomotion protocol takes 21 h, including cell suspension preparation, optimized bacterial attachment to functionalized cantilever, and nanomotion recording before and after antibiotic exposure. We applied this protocol to MTB isolates (n = 40) and were able to discriminate between susceptible and resistant strains for INH and RIF with a maximum sensitivity of 97.4% and 100%, respectively, and a maximum specificity of 100% for both antibiotics when considering each nanomotion recording to be a distinct experiment. Grouping recordings as triplicates based on source isolate improved sensitivity and specificity to 100% for both antibiotics. Nanomotion technology can potentially reduce time-to-result significantly compared to the days and weeks currently needed for current phenotypic ASTs for MTB. It can further be extended to other anti-TB drugs to help guide more effective TB treatment.

Determination of nutrient and toxic elements in food reference materials by suspension preparation and TXRF analysis

There is a need to maintain nutrient element adequacy through food consumption. Due to the risk of toxic element exposure during food consumption, an accurate and precise characterisation approach is required. This makes the development of a simpler and faster procedure a great concern, especially in Indonesia, where time-consuming methods such as atomic absorption spectroscopy (AAS) and inductively coupled plasma-mass spectrometry (ICP-MS) are still dominant. Therefore, ththe present work aimed to evaluate a suspension technique for food matrices in combination with TXRF S4-TSTAR (total reflection X-ray fluorescence) in Indonesia. The examination focused on standard reference materials (SRM) such as 1570a spinach leaves (SL), 1548 typical diet (TD), 1566b oyster tissue (OT), and 8418 bovine muscle (BM). The concentration of elements was determined by mixing each SRM with internal standard gallium (Ga). This was followed by the comparison of the concentration and sensitivity of Ga to each element in SRM. The TXRF performance was evaluated by quantifying nutrients such as calcium (Ca), iron (Fe), zinc (Zn), copper (Cu), manganese (Mn), and toxic element namely arsenic (As). The trueness and precision were calculated through %bias, %recovery, coefficient of variance (%CV), and HorRat ratio (r). The recovery of all elements in SRMs was within 81.20 - 103.35%, except for SRM TD and BM which were 61.73 - 91.70 and 73.78 - 99.41%, respectively. The CV of all SRMs was within the range of 0.63 - 9.54%, except for SRM BM and HorRat ratios which were 2.10 - 25.22 and 0.12 - 3.21%. Based on the results, the concentration of Zn was in good agreement with the primary method of neutron activation analysis (NAA). This showed that TXRF S4-TSTAR had good trueness and precision on SRM food matrices, and could be a promising method to be applied for element characterisation in Indonesian nutritional research.

The problems of unifying information in the patient leaflets in Ukraine during the COVID-19 pandemic and wartime

The information in the patient leaflets on the medicines must be unified, have a good structure and be clearly presented to the patient for prevent information asymmetry. We substantiated and worked a methodic of comparative analysis of antibiotic using for example of 74 instructions on the drug of one INN, which are registered on the markets of Ukraine, EU countries and Japan. Ways to unify information in to improve patient awareness of the correct use of drugs, especially antibiotics, are substantiated and presented.
 Aim – to conduct a comparative analysis of the patient leaflets for antibacterial medicines and determine the presence or absence of information asymmetry using the example of antibiotics of one INN that are registrated on the Ukrainian market or they received as a humanitarian aid from Europe in wartime, in order to establish inconsistencies in the information structure and data in the information leaflets.
 The objects were information leaflets for antibiotics: azithromycin, amoxiclav, amoxicillin, zinnate, cefazolin, cefatoxin. Methods were informational, comparative and content analysis. Methodology of information asymmetry analysis using the example of information leaflets for the medicine azithromycin from various manufacturers available on the market of Ukraine.
 As a result, significant differences were established in 3 sections: «Indications», «Peculiarities of use», suspension preparation, etc. Significant differences were found in the information leaflets for prescribing azithromycin for skin infections, which requires immediate unification in the information leaflets of data for correct prescribing by a doctor using an electronic prescription and good use for a treatment course. It was established that in the for azithromycin from one manufacturer, which is on the market of Ukraine and EU countries, significant differences in information were found in 10 sections of the information leaflets, which requires the unification of information accordingly.
 We have reasoned that the information leaflets should contain information that is important for the correct use of the medicine, in order to prevent potentially dangerous consequences that may arise from incorrect preparation or dosage. We grounded the information leaflets should include a section on the disposal of expired or unused medicines to harmonize with the information leaflets іn the EU countries.

The Design and Verification of a TBP/OK Pyrolysis Incineration Stable System

This project designed and created a large-scale pyrolysis incineration system that decomposes waste tributyl phosphate/kerosene (TBP/OK) solvent from the plutonium/uranium process (PU-REX) in the nuclear industry. This system consists of a suspension preparation system (SPS), a pyrolysis and ash discharge system (PADS), and a combustion furnace and gas treatment system (CGS). The preparation of a waste TBP/OK emulsified suspension in the SPS, continuous operation experiments of the PADS and CGS, and pyrolysis incineration experiments were conducted. The results show that the SPS achieved the automatic preparation and feeding of emulsified suspensions at 30 kg/h. The PADS decomposed 99.99% of the TBP/OK emulsified suspensions and fixed more than 99% of phosphorus. The CGS burned the pyrolysis gas without diesel oil and purified more than 99.995% of the radioactive nuclides. This system showed great feasibility, stability, continuity, and safety, proving the compatibility between multiple systems of this new design. This paper marks a significant step in pyrolysis incineration technology for the nuclear industry.

Preparation of High-Viability Single-Cell Suspensions from African Turquoise Killifish Brain Tissue.

Studying the brain at the single-cell level has become increasingly popular in recent years. This, however, remains challenging, especially in emerging model organisms. To carry out single-cell sequencing, the preparation of a high-viability single-cell suspension is critical. In this protocol, we describe how to prepare a high-viability single-cell suspension starting from brain tissue of the African turquoise killifish (Nothobranchius furzeri). The protocol consists of dissection, enzymatic and mechanical dissociation of the brain tissue, and debris removal. The protocol described here has been successfully used for 10× Genomics single-cell sequencing of the telencephalon of adult killifish, which requires a cell viability of at least 70%. In addition to single-cell sequencing experiments, the single-cell suspension generated can be used for other applications, including cell culture and flow cytometry.

Chromosome Analysis and Sorting Using Conventional Flow Cytometers.

The fluorescent dyes Hoechst (HO) and Chromomycin A3 (CA3) are commonly used for bivariate flow karyotyping to distinguish individual chromosomes from one another based on differences in base composition and DNA content. However, analysis of chromosomes using this fluorescent dye combination requires a flow cytometer equipped with lasers of specific wavelengths and higher power than is typical of conventional flow cytometers. This unit presents a chromosome staining technique with a dye combination of DAPI and propidium iodide (PI). Chromosomes stained using this dye combination can be analyzed on conventional flow cytometers equipped with a typical configuration of lasers and optics. © 2023 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Cell culture and metaphase harvest of suspension cell line Alternate Protocol 1: Cell culture and metaphase harvest of adherent cell line Basic Protocol 2: Preparation of chromosome suspension using polyamine isolation buffer Basic Protocol 3: Staining chromosomes with DAPI and propidium iodide Alternate Protocol 2: Staining chromosomes with Hoechst and Chromomycin A3 Basic Protocol 4: Bivariate flow karyotyping on a cell analyzer Basic Protocol 5: Bivariate flow karyotyping on a cell sorter Basic Protocol 6: Purification of flow-sorted chromosomes.

A Mouse Model of MC903-Induced Atopic Dermatitis.

Atopic dermatitis (AD) is a chronic, relapsing, and extremely pruritic inflammatory skin disease with a particular impact on children. AD pathogenesis is not yet fully understood, and there is no curative treatment for this disease. Therefore, several genetically or chemically-induced AD mouse models have been developed. These preclinical mouse models are an indispensable research tool for studying AD pathogenesis and evaluating the efficacy of new candidate AD therapeutics. A commonly used mouse model of AD has been developed using the topical application of a low-calcemic analog of vitamin D3, MC903, to induce AD-like inflammatory phenotypes that closely resemble human AD. Moreover, this model shows a minimal effect on systemic calcium metabolism that is observed in the vitamin D3-induced AD model. Thus, an expanding number of studies use the MC903-induced AD model to interrogate AD pathobiology in vivo and to test new candidate small molecule and monoclonal antibody therapies. This protocol describes in detail functional measurements including the measurement of skin thickness, which is a surrogate marker for ear skin inflammation, as well as itch assessment, histological evaluation to assess the structural changes associated with AD skin inflammation, and preparation of single-cell suspensions from ear skin and draining lymph nodes for the assessment of inflammatory leukocyte subset infiltration in these tissues using flow cytometry. © 2023 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol: Topical application of MC903 induces AD-like skin inflammation Support Protocol 1: Measurement of ear skin thickness Support Protocol 2: Itch assessment Support Protocol 3: Dissection of ear skin and ear draining lymph nodes Support Protocol 4: Histological evaluation and quantification Support Protocol 5: Preparation of single-cell suspension from ear skin and draining lymph nodes for the assessment of inflammatory immune cell infiltration using flow cytometry.

Testing of the Efficacy of Bee Probiotic Lactobacilli Under In Vivo Conditions

Abstract American foulbrood is amongst the most dangerous diseases of the bee-family affecting many honeybee colonies worldwide. In countries of European Union, based on veterinary legislation, the bee colonies tested positive to American foulbrood are eradicated with high economic losses. It is therefore necessary to look for effective prevention, especially by the using of natural ingredients such as probiotics. In this study, we used lactobacilli isolated from digestive tracts of adult healthy honey bees and selected based on their good probiotic properties and ability to inhibit the growth of Paenibacillus larvae. These isolates were identified as Lactobacillus brevis and Lactobacillus plantarum. Night cultures of both strains were used for the preparation of probiotic suspensions and pollen was selected as an appropriate carrier for application of probiotic lacto-bacilli to the hives. Half a litre of pollen suspension was prepared for each hive. The suspension for experimental hives contained probiotic lactobacilli in concentration of 107–108 CFU.ml−1. Bees in control hives received pollen suspension without addition of probiotic lacto-bacilli. The suspensions were supplied to bees three times, once a week. Before and during administration of probiotics, samples of honey bees from each hive were taken every week and numbers of lactobacilli, P. larvae, enterobacteria and coliform bacteria were determined in their digestive tracts. Four weeks after the first administration of probiotic-pollen solution the numbers of lactobacilli were increased approximately by 0.5 log. Before starting this experiment, P. larvae (approx. 107 CFU.ml−1) were detected in digestive tracts of honey bees. In the experimental group after 3 weeks and in the control group after 4 weeks, no viable counts of P. larvae were found. The numbers of enterobacteria, coliform bacteria and Bacillus sp. decreased in both groups. During the experiment the health and condition of the hives were monitored. In addition, also monitored were: the development of the bee colony, the number of dead bees, the amount of hive debris, the aggressiveness of the bees, and the amount of honey spun; later during the autumn treatment also the fall of Varroa jacobsoni was noted. In the experimental and control groups, we recorded a decrease in the amount of hive debris. Only in the experimental group was a slightly above-average development of the bee colony, slightly above-average honey yields and a 70 % lower drop of Varroa mites. Based on these results, we assumed that the probiotic-pollen solution had a positive influence on the composition of microbiota in bee digestive tracts and it can increase resistance to P. larvae. It also had a positive effect on the health and condition of the bee colony. Probiotic-free pollen solution showed similar but weaker effects.

Synthesis of Citrate-Capped Sn Nanoparticles with Excellent Oxidation Resistance for High-Performance Electrically Conductive Adhesives

Tin (Sn) metallic material is one of the best-known metals and has been extensively studied due to its ease of processing, low melting point, and low cost. However, it is still challenging to synthesize high-processing performance and antioxidant Sn nanoparticles with tunable sizes. This research reported a facile, easy-to-scale-up polyol-mediated synthesis of Sn nanoparticles assisted by sodium citrates as a capping agent. The presence of citrate capping facilitated uniform nucleation of Sn nanoparticles and enhanced their antioxidant capacity and dispersion properties. These nanoparticles can remain stable against oxidation for more than 270 days in an ambient atmosphere, even under continuous heating at 200 °C for over 12 h. The citrate capping also inhibits interparticle agglomeration and allows the preparation of high-quality suspensions in water and many conventional organics. Moreover, efficient size tuning over a wide range (60 nm–1 μm) can be achieved simply by changing the Sn2+ precursor concentrations. The above-mentioned antioxidant capacity and processability allow them to combine with silver flakes in thermosetting epoxy resin as complementary conductive fillers effectively, creating conductive pathways among the silver flakes to obtain high-performance electrically conductive adhesives (ECAs). By adding Sn nanoparticles as complementary conductive fillers, the resistivity of the ECAs can be reduced to 1/6000 of that of the ECAs filled with only the same mass ratio of silver flakes, exhibiting its great potential in future electronics.

Single-Cell Suspension Preparation from Nile Tilapia Intestine for Single-Cell Sequencing.

Nile tilapia is one of the most commonly cultured freshwater fish species worldwide and is a widely used research model for aquaculture fish studies. The preparation of high-quality single-cell suspensions is essential for single-cell level studies such as single-cell RNA or genome sequencing. However, there is no ready-to-use protocol for aquaculture fish species, particularly for the intestine of tilapia. The effective dissociation enzymes vary depending on the tissue type. Therefore, optimizing the tissue dissociation protocol by selecting the appropriate enzyme or enzyme combination to obtain enough viable cells with minimum damage is essential. This study illustrates an optimized protocol to prepare a high-quality single-cell suspension from Nile tilapia intestine with an enzyme combination of collagenase/dispase. This combination is highly effective for dissociation with the utilization of bovine serum albumin and DNase to reduce cell aggregation after digestion. The cell output satisfies the requirements for single-cell sequencing, with 90% cell viability and a high cell concentration. This protocol can also be modified to prepare a single-cell suspension from the intestines of other fish species. This research provides an efficient reference protocol and reduces the need for additional trials in the preparation of single-cell suspensions for aquaculture fish species.

Facile Technology for Extemporaneous Preparation of Long-Acting Injectable Microparticulate Suspensions at the Patient Side

In this study, we present an innovative and facile in situ approach for extemporaneous preparation of sterile microparticles. An amazingly simple approach, in situ technology circumvents the stability, and scale up challenges as well as sterilization issues associated with long-acting particulate systems. Monophasic preconcentrates of donepezil base (DPZ), a model drug with a biodegradable polymer poly (DL-lactide-co-glycolide) (PLGA), with stabilizer were prepared by simple solution and sterilized by filtration (0.22 micron). The sterile preconcentrates when added to aqueous dextrose solution (total volume < 3mL) generated ready-to-inject DPZ PLGA microparticles (DPZ-PLGA-MP) with high reproducibility, entrapment efficiency (> 80%), and size ~ 80 micron. DPZ micro suspension(DPZ-MS) with high precipitation efficiency (> 90%) and size ~ 80 micron was obtained in a similar manner omitting PLGA. XRD and DSC study confirmed decreased crystallinity in the presence of PLGA. No interaction between PLGA and DPZ was evident in the FTIR study. The microparticulate dispersions exhibited good in vitro injectability when tested using the texture analyzer (force < 5 N). When evaluated using the dialysis bag method (Himedia 12-14kDa molecular weight cutoff), both microparticulate formulations exhibited controlled release up to 1week in vitro. Further, low burst release of ~ 10% at the end of 6h in the ex vivo chicken muscle study proposes great promise. Our data propose the facile extemporaneous generation of microparticles as a practical and promising approach for development of long-acting injectables. This facile approach could serve as platform technology for other drug candidates.

Preparation of a Non-Cardiomyocyte Cell Suspension for Single-Cell RNA Sequencing from a Post-Myocardial Infarction Adult Mouse Heart.

Myocardial infarction (MI) is one of the most common cardiovascular diseases, with increasing mortality worldwide. Non-cardiomyocytes account for more than half of the total cardiac cell population, and they contribute to adaptive compensations upon myocardial injury, including inflammatory responses, tissue repair, and scar formation. To study the post-MI cardiac microenvironment, single-cell RNA sequencing (scRNA-seq) is widely used to identify different cardiac cell types and intercellular communications. Among the procedures of scRNA-seq sample preparation, preparing the cell suspension is one of the most critical steps, because the cell viability can affect the quality of the scRNA-seq results. Therefore, we designed an experimental protocol for preparing a non-cardiomyocyte cell suspension from post-MI mouse hearts with an extra focus on improving the cell viability by choosing mild digestive enzymes, controlling the digestion time, and applying fluorescence-activated cell sorting (FACS). Finally, we isolated CD45+ cells from the non-cardiomyocyte cell suspension obtained through this protocol, and then we performed scRNA-seq.

On-site determination of water toxicity based on freeze-dried electrochemically active bacteria

Our previous studies have reported water toxicity determination with a fresh electrochemically active bacteria (EAB) suspension as the sensing element, which exhibits high sensitivity and has great prospects in providing early warning about water pollution. However, because the preparation of fresh EAB suspensions is time-consuming, these studies are not suitable for the on-site determination of water toxicity. To solve this problem, this study investigated the rapid preparation of an EAB suspension by the rehydration of freeze-dried EABs and established a novel method for the on-site determination of water toxicity based on the freeze-dried EAB model strain Shewanella oneidensis MR-1. The results demonstrate that the optimal cryoprotectant for S. oneidensis MR-1 freeze drying is 7.5 % (w/v) skimmed milk powder. Compared with fresh S. oneidensis MR-1, freeze-dried S. oneidensis MR-1 exhibits similar extracellular electron transfer (EET) performance (74.7 % ± 0.3 %) and slightly lower sensitivity for water toxicity determination (65.8 % ± 2.2 %) with the optimal cryoprotectant. On-site determination of water toxicity was realized by using freeze-dried S. oneidensis MR-1, and the detection limits of five common toxic pollutants (Cd2+, Pb2+, Cu2+, phenol and dichlorophenol) reached 0.5 mg/L. Water toxicity determination is capable of resisting common interferences, e.g., glucose, lactate, nitrate and nitrite, and shows high accuracy in practical applications.

METHOD OF OBTAINING WATER-COAL SUSPENSION AND HEAT AND MASS EXCHANGE EQUIPMENT FOR ITS APPLICATION

The analysis of methods and equipment for obtaining water-coal suspensions was carried out. The purpose of the research is to improve the quality of the resulting suspension due to more effective grinding and increasing the amount of its solid component. Heat and mass exchange equipment has been developed, with the help of which it is possible to achieve an increase in the efficiency of hydraulic impact due to an increase in the frequency of pulsations when obtaining water-coal suspensions. The use of water-coal suspensions as an energy fuel makes it possible to improve the technical, economic and environmental performance of power plants. Thus, during the conversion of the boiler to water-coal fuel, it was possible to increase the thermal power of the boiler from 13,2 to 21,1 MW when replacing 70 % of coal with water-coal fuel. Currently, there are different methods and equipment for the preparation of water-coal suspension. However, they all have certain drawbacks. The main disadvantages of the method of obtaining water-coal suspensions are multi-stage, metal consumption, difficulty of installation and start-up, high energy costs, and the resulting suspension is of insufficiently high quality. Also, the disadvantages are the limited frequency of hydraulic shock pulses during processing in one cycle, which is insufficient for the preparation of highly concentrated suspensions that meet the requirements for liquid composite fuel intended for direct combustion. In order to improve the quality of the resulting suspension due to more effective grinding and increasing the amount of its solid component, heat and mass exchange equipment was developed, with the help of which it is possible to achieve an increase in the efficiency of the hydraulic impact action due to an increase in the frequency of pulsations during the production of water-coal suspensions. As heat and mass exchange equipment, rotary pulsation devices (RPD) operating according to the method of discrete-pulse energy input (DPEI) were used. The DPEI method involves the creation of a large number of working bodies or working elements evenly distributed in the water, which transform stationary thermal, mechanical or other types of energy into energetically powerful pulses, discrete in time and space.

Functional Phenotyping of Lung Mouse CD4+ T Cells Using Multiparametric Flow Cytometry Analysis.

Gammaherpesviruses such as Epstein-Barr virus (EBV) are major modulators of the immune responses of their hosts. In the related study (PMID: 35857578), we investigated the role for Ly6Chi monocytes in shaping the function of effector CD4+ T cells in the context of a murine gammaherpesvirus infection (Murid gammaherpesvirus 4) as a model of human EBV. In order to unravel the polyfunctional properties of CD4+ T-cell subsets, we used multiparametric flow cytometry to perform intracellular staining on lung cells. As such, we have developed herein an intracellular staining workflow to identify on the same samples the cytotoxic and/or regulatory properties of CD4+ lymphocytes at the single-cell level. Briefly, following perfusion, collection, digestion, and filtration of the lung to obtain a single-cell suspension, lung cells were cultured for 4 h with protein transport inhibitors and specific stimulation media to accumulate cytokines of interest and/or cytotoxic granules. After multicolor surface labeling, fixation, and mild permeabilization, lung cells were stained for intracytoplasmic antigens and analyzed with a Fortessa 4-laser cytometer. This method of quantifying cytotoxic mediators as well as pro- or anti-inflammatory cytokines by flow cytometry has allowed us to decipher at high resolution the functional heterogeneity of lung CD4+ T cells recruited after a viral infection. Therefore, this analysis provided a better understanding of the importance of CD4+ T-cell regulation to prevent the development of virus-induced immunopathologies in the lung. Key features • High-resolution profiling of the functional properties of lung-infiltrating CD4+ T cells after viral infection using conventional multiparametric flow cytometry. • Detailed protocol for mouse lung dissection, preparation of single-cell suspension, and setup of multicolor surface/intracellular staining. • Summary of optimal ex vivo restimulation conditions for investigating the functional polarization and cytokine production of lung-infiltrating CD4+ T cells. • Comprehensive compilation of necessary biological and technical controls to ensure reliable data analysis and interpretation.