Effective Carbon Dioxide Mitigation and Improvement of Compost Nutrients with the Use of Composts' Biochar.

Composting is a process that emits environmentally harmful gases: CO2, CO, H2S, and NH3, negatively affecting the quality of mature compost. The addition of biochar to the compost can significantly reduce emissions. For effective CO2 removal, high doses of biochar (up to 20%) are often recommended. Nevertheless, as the production efficiency of biochar is low-up to 90% mass loss-there is a need for research into the effectiveness of lower doses. In this study, laboratory experiments were conducted to observe the gaseous emissions during the first 10 days of composting with biochars obtained from mature composts. Biochars were produced at 550, 600, and 650 °C, and tested with different doses of 0, 3, 6, 9, 12, and 15% per dry matter (d.m.) in composting mixtures, at three incubation temperatures (50, 60, and 70 °C). CO2, CO, H2S, and NH3 emissions were measured daily. The results showed that the biochars effectively mitigate CO2 emissions during the intensive phase of composting. Even 3-6% d.m. of compost biochars can reduce up to 50% of the total measured gas emissions (the best treatment was B650 at 60 °C) and significantly increase the content of macronutrients. This study confirmed that even low doses of compost biochars have the potential for enhancing the composting process and improving the quality of the material quality.

Application of Composts' Biochar as Potential Sorbent to Reduce VOCs Emission during Kitchen Waste Storage.

It is expected that due to the new European Union regulation focus on waste management, managing kitchen waste will become more important in the future, especially in households. Therefore, it is crucial to develop user-friendly and odour-free containers to store kitchen waste. The study aimed to test the effectiveness of composts' biochar in reducing noxious odours and volatile organic compounds (VOCs) released during kitchen waste storage. Various amounts of compost biochar (0%, 1%, 5%, and 10%) were added to food waste samples and incubated for seven days at 20 °C. The released VOCs were analysed on days 1, 3, and 7 of the storage simulation process. The results indicated that adding 5-10% of composts' biochar to kitchen waste significantly reduced the emissions in 70% of the detected VOCs compounds. Furthermore, composts' biochar can be used to eliminate potential odour components and specific dangerous VOCs such as ethylbenzene, o-xylene, acetic acid, and naphthalene. A new composts' biochar with a unique composition was particularly effective in reducing VOCs and could be an excellent solution for eliminating odours in kitchen waste containers.

Carbon Monoxide Production during Bio-Waste Composting under Different Temperature and Aeration Regimes.

Despite the development of biorefinery processes, the possibility of coupling the "conventional" composting process with the production of biochemicals is not taken into account. However, net carbon monoxide (CO) production has been observed during bio-waste composting. So far, O2 concentration and temperature have been identified as the main variables influencing CO formation. This study aimed to investigate CO net production during bio-waste composting under controlled laboratory conditions by varying aeration rates and temperatures. A series of composting processes was carried out in conditions ranging from mesophilic to thermophilic (T = 35, 45, 55, and 65 °C) and an aeration rate of 2.7, 3.4, 4.8, and 7.8 L·h-1. Based on the findings of this study, suggestions for the improvement of CO production throughout the composting process have been developed for the first time. The highest concentrations of CO in each thermal variant was achieved with an O2 deficit (aeration rate 2.7 L·h-1); additionally, CO levels increased with temperature, reaching ~300 ppm at 65 °C. The production of CO in mesophilic and thermophilic conditions draws attention to biological CO formation by microorganisms capable of producing the CODH enzyme. Further research on CO production efficiency in these thermal ranges is necessary with the characterization of the microbial community and analysis of the ability of the identified bacteria to produce the CODH enzyme and convert CO from CO2.

Calcification alters the viscoelastic properties of tendon fascicle bundles depending on matrix content.

Tendon fascicle bundles are often used as biological grafts and thus must meet certain quality requirements, such as excluding calcification, which alters the biomechanical properties of soft tissues. In this work, we investigate the influence of early-stage calcification on the mechanical and structural properties of tendon fascicle bundles with varying matrix content. The calcification process was modeled using sample incubation in concentrated simulated body fluid. Mechanical and structural properties were investigated using uniaxial tests with relaxation periods, dynamic mechanical analysis, as well as magnetic resonance imaging and atomic force microscopy. Mechanical tests showed that the initial phase of calcification causes an increase in the elasticity, storage, and loss modulus, as well as a drop in the normalized value of hysteresis. Further calcification of the samples results in decreased modulus of elasticity and a slight increase in the normalized value of hysteresis. Analysis via MRI and scanning electron microscopy showed that incubation alters fibrillar relationships within the tendon structure and the flow of body fluids. In the initial stage of calcification, calcium phosphate crystals are barely visible; however, extending the incubation time for the next 14 days results in the appearance of calcium phosphate crystals within the tendon structure and leads to damage in its structure. Our results show that the calcification process modifies the collagen-matrix relationships and leads to a change in their mechanical properties. These findings will help to understand the pathogenesis of clinical conditions caused by calcification process, leading to the development of effective treatments for these conditions. STATEMENT OF SIGNIFICANCE: This study investigates how calcium mineral deposition in tendons affects their mechanical response and which processes are responsible for this phenomenon. By analyzing the elastic and viscoelastic properties of animal fascicle bundles affected by calcification induced via incubation in concentrated simulated body fluid, the study sheds light on the relationship between structural and biochemical changes in tendons and their altered mechanical response. This understanding is crucial for optimizing tendinopathy treatment and preventing tendon injury. The findings provide insights into the calcification pathway and its resulting changes in the biomechanical behaviors of affected tendons, which have been previously unclear.

Biological treatment of biowaste as an innovative source of CO-The role of composting process.

Carbon monoxide (CO) is an essential "building block" for producing everyday chemicals on industrial scale. Carbon monoxide can also be generated though a lesser-known and sometimes forgotten biorenewable pathways that could be explored to advance biobased production from large and more sustainable sources such as bio-waste treatment. Organic matter decomposition can generate carbon monoxide both under aerobic and anaerobic conditions. While anaerobic carbon monoxide generation is relatively well understood, the aerobic is not. Yet many industrial-scale bioprocesses involve both conditions. This review summarizes the necessary basic biochemistry knowledge needed for realization of initial steps towards biobased carbon monoxide production. We analyzed for the first time, the complex information about carbon monoxide production during aerobic, anaerobic bio-waste treatment and storage, carbon monoxide-metabolizing microorganisms, pathways, and enzymes with bibliometric analysis of trends. The future directions recognizing limitations of combined composting and carbon monoxide production have been discussed in greater detail.

Influence of blood sample storage and different types of anticoagulants on results of NK cytotoxicity tests.

Introduction: Natural killer (NK) cells are important players in the human immune response. Impaired NK function may lead to serious, life-threatening conditions. Defects may be consequences of genetic mutations or results of secondary factors such as infections, malignancies and autoimmune diseases. The cytotoxicity test is very useful, but its accessibility is limited to special immunological laboratories. Blood samples are often transported to remote centers, which takes time and requires special conditions.The aim of this study was to compare cytotoxicity assay results between samples preserved with three different anticoagulants to standardize the diagnostic procedure. Material and methods: Peripheral blood from healthy donors was taken with three anticoagulants: heparin, K2EDTA and citrate. Peripheral blood mononuclear cells (PBMC) were isolated and tested directly after blood drawing and after 24-hour storage. Cytotoxic abilities of NK cells were tested in 4 h co-culture with K562. NK cytotoxicity was measured by flow cytometry. Results: In most cases of analyzed healthy donors, cytotoxicity results were similar regardless of type of anticoagulant. However, the highest mean values were obtained in samples with citrate. There was a significant decrease in cytotoxicity after 24 hours of storage of the whole blood at ambient temperature. The mean drop in cytotoxicity results was substantial for all anticoagulants: 76% for heparin, 67% for citrate and 70% for EDTA. Conclusions: Results of spontaneous NK cytotoxicity seem to be affected by the anticoagulants used for blood protection. Commercial instant cytotoxicity testing and delayed analysis after blood storage gave the highest results in blood with sodium citrate.

Electrospun Membrane Surface Modification by Sonocoating with HA and ZnO:Ag Nanoparticles-Characterization and Evaluation of Osteoblasts and Bacterial Cell Behavior In Vitro.

Guided tissue regeneration and guided bone regeneration membranes are some of the most common products used for bone regeneration in periodontal dentistry. The main disadvantage of commercially available membranes is their lack of bone cell stimulation and easy bacterial colonization. The aim of this work was to design and fabricate a new membrane construct composed of electrospun poly (D,L-lactic acid)/poly (lactic-co-glycolic acid) fibers sonocoated with layers of nanoparticles with specific properties, i.e., hydroxyapatite and bimetallic nanocomposite of zinc oxide-silver. Thus, within this study, four different variants of biomaterials were evaluated, namely: poly (D,L-lactic acid)/poly (lactic-co-glycolic acid) biomaterial, poly(D,L-lactic acid)/poly (lactic-co-glycolic acid)/nano hydroxyapatite biomaterial, poly (D,L-lactic acid)/poly (lactic-co-glycolic acid)/nano zinc oxide-silver biomaterial, and poly (D,L-lactic acid)/poly (lactic-co-glycolic acid)/nano hydroxyapatite/nano zinc oxide-silver biomaterial. First, it was demonstrated that the wettability of biomaterials-a prerequisite property important for ensuring desired biological response-was highly increased after the sonocoating process. Moreover, it was indicated that biomaterials composed of poly (D,L-lactic acid)/poly (lactic-co-glycolic acid) with or without a nano hydroxyapatite layer allowed proper osteoblast growth and proliferation, but did not have antibacterial properties. Addition of a nano zinc oxide-silver layer to the biomaterial inhibited growth of bacterial cells around the membrane, but at the same time induced very high cytotoxicity towards osteoblasts. Most importantly, enrichment of this biomaterial with a supplementary underlayer of nano hydroxyapatite allowed for the preservation of antibacterial properties and also a decrease in the cytotoxicity towards bone cells, associated with the presence of a nano zinc oxide-silver layer. Thus, the final structure of the composite poly (D,L-lactic acid)/poly (lactic-co-glycolic acid)/nano hydroxyapatite/nano zinc oxide-silver seems to be a promising construct for tissue engineering products, especially guided tissue regeneration/guided bone regeneration membranes. Nevertheless, additional research is needed in order to improve the developed construct, which will simultaneously protect the biomaterial from bacterial colonization and enhance the bone regeneration properties.

Rehydration of the Tendon Fascicle Bundles Using Simulated Body Fluid Ensures Stable Mechanical Properties of the Samples.

In this work, we investigate the influence of dehydration and subsequent rehydration of tendon fascicle bundles on their structural and mechanical properties by using distilled water, 0.9% NaCl, 10% NaCl, SBF, and double concentrated SBF (SBFx2). The properties of tendon fascicle bundles were investigated by means of uniaxial tests with relaxation periods and hysteresis for samples with various interfascicular matrix content, dissected from the anterior and posterior areas of bovine tendon. Uniaxial tests with relaxation periods and analysis of sample geometry and weight showed that dehydration alters the modulus of elasticity dependent on the interfascicular matrix content and influences the viscoelastic properties of tendon fascicle bundles. Tensile and relaxation tests revealed that changes resulting from excessive sample drying can be reversed by rehydration in an SBF bath solution for elastic strain range above the toe region. Rehydration in SBF solution led to minor differences in mechanical properties when compared to control samples. Moreover, anterior samples with greater interfascicular matrix content, despite their lower stiffness, are less sensitive to sample drying. The obtained results allow us to limit the discrepancies in the measurement of mechanical properties of wet biological samples and can be useful to researchers investigating soft tissue mechanics and the stability of transplant materials.

Aerobic Biostabilization of the Organic Fraction of Municipal Solid Waste-Monitoring Hot and Cold Spots in the Reactor as a Novel Tool for Process Optimization.

The process of aerobic biostabilization (AB) has been adopted for treatment of the organic fraction of municipal solid waste (OFMSW). However, thermal gradients and some side effects in the bioreactors present difficulties in optimization of AB. Forced aeration is more effective than natural ventilation of waste piles, but "hot and cold spots" exist due to inhomogeneous distribution of air and heat. This study identified the occurrence of hot and cold spots during the OFMSW biostabilization process at full technical scale. It was shown that the number of hot and cold spots depended on the size of the pile and aeration rate. When the mass of stabilized waste was significantly lower and the aeration rate was two-fold higher the number of anaerobic hot spots decreased, while cold spots increased. In addition, the results indicated that pile construction with sidewalls decreased the number of hot spots. However, channelizing the airflow under similar conditions increased the number of cold spots. Knowledge of the spatial and temporal distribution of process gases can enable optimization and adoption of the OFMSW flow aeration regime. Temperature monitoring within the waste pile enables the operator to eliminate undesirable "hot spots" by modifying the aeration regime and hence improve the overall treatment efficiency.

The dispersion of viscoelastic properties of fascicle bundles within the tendon results from the presence of interfascicular matrix and flow of body fluids.

In this work, we investigate differences in the mechanical and structural properties of tendon fascicle bundles dissected from different areas of bovine tendons. The properties of tendon fascicle bundles were investigated by means of uniaxial tests with relaxation periods and hysteresis, dynamic mechanical analysis (DMA), as well as magnetic resonance imaging (MRI). Uniaxial tests with relaxation periods revealed greater elastic modulus, hysteresis, as well as stress drop during the relaxation of samples dissected from the posterior side of the tendon. However, the normalized stress relaxation curves did not show a statistically significant difference in the stress drop between specimens cut from different zones or between different strain levels. Using dynamic mechanical analysis, we found that fascicle bundles dissected from the anterior side of the tendon had lower storage and loss moduli, which could result from altered fluid flow within the interfascicular matrix (IFM). The lower water content, diffusivity, and higher fractional anisotropy of the posterior part of the tendon, as observed using MRI, indicates a different structure of the IFM, which controls the flow of fluids within the tendon. Our results show that the viscoelastic response to dynamic loading is correlated with fluid flow within the IFM, which was confirmed during analysis of the MRI results. In contrast to this, the long-term relaxation of tendon fascicle bundles is controlled by viscoplasticity of the IFM and depends on the spatial distribution of the matrix within the tendon. Comparison of results from tensile tests, DMA, and MRI gives new insight into tendon mechanics and the role of the IFM. These findings may be useful in improving the diagnosis of tendon injury and effectiveness of medical treatments for tendinopathies.

Mesenchymal Stem Cells for Neurological Disorders.

Neurological disorders are becoming a growing burden as society ages, and there is a compelling need to address this spiraling problem. Stem cell-based regenerative medicine is becoming an increasingly attractive approach to designing therapies for such disorders. The unique characteristics of mesenchymal stem cells (MSCs) make them among the most sought after cell sources. Researchers have extensively studied the modulatory properties of MSCs and their engineering, labeling, and delivery methods to the brain. The first part of this review provides an overview of studies on the application of MSCs to various neurological diseases, including stroke, traumatic brain injury, spinal cord injury, multiple sclerosis, amyotrophic lateral sclerosis, Alzheimer's disease, Huntington's disease, Parkinson's disease, and other less frequently studied clinical entities. In the second part, stem cell delivery to the brain is focused. This fundamental but still understudied problem needs to be overcome to apply stem cells to brain diseases successfully. Here the value of cell engineering is also emphasized to facilitate MSC diapedesis, migration, and homing to brain areas affected by the disease to implement precision medicine paradigms into stem cell-based therapies.

Neuroinflammation evoked by brain injury in a rat model of lacunar infarct.

Stroke is the leading cause of long-term, severe disability worldwide. Immediately after the stroke, endogenous inflammatory processes are upregulated, leading to the local neuroinflammation and the potentiation of brain tissue destruction. The innate immune response is triggered as early as 24 h post-brain ischemia, followed by adaptive immunity activation. Together these immune cells produce many inflammatory mediators, i.e., cytokines, growth factors, and chemokines. Our study examines the immune response components in the early stage of deep brain lacunar infarct in the rat brain, highly relevant to the clinical scenario. The lesion was induced by stereotactic injection of ouabain into the adult rat striatum. Ouabain is a Na/K ATPase pump inhibitor that causes excitotoxicity and brings metabolic and structural changes in the cells leading to focal brain injury. We have shown a surge of neurodegenerative changes in the peri-infarct area in the first days after brain injury. Immunohistochemical analysis revealed early microglial activation and the gradual infiltration of immune cells with a significant increase of CD4+ and CD8+ T lymphocytes in the ipsilateral hemisphere. In our studies, we identified the higher level of pro-inflammatory cytokines, i.e., interleukin-1α, interleukin-1ß, tumor necrosis factor-α, and interferon-γ, but a lower level of anti-inflammatory cytokines, i.e., interleukin-10 and transforming growth factor-ß2 in the injured brain than in normal rats. Concomitantly focal brain injury showed a significant increase in the level of chemokines, i.e., monocyte chemoattractant protein-1 and CC motif chemokine ligand 5 compared to control. Our findings provide new insights into an early inflammatory reaction in our model of the deep-brain lacunar infarct. The results of this study may highlight future stroke immunotherapies for targeting the acute immune response accompanied by the insult.

A 3D Scanning System for Inverse Analysis of Moist Biological Samples: Design and Validation Using Tendon Fascicle Bundles.

In this article, we present the design and validation of a non-contact scanning system for the development of a three-dimensional (3D) model of moist biological samples. Due to the irregular shapes and low stiffness of soft tissue samples, the use of a non-contact, reliable geometry scanning system with good accuracy and repeatability is required. We propose a reliable 3D scanning system consisting of a blue light profile sensor, stationary and rotating frames with stepper motors, gears and a five-phase stepping motor unit, single-axis robot, control system, and replaceable sample grips, which once mounted onto the sample, are used for both scanning and mechanical tests. The proposed system was validated by comparison of the cross-sectional areas calculated based on 3D models, digital caliper, and vision-based methods. Validation was done on regularly-shaped samples, a wooden twig, as well as tendon fascicle bundles. The 3D profiles were used for the development of the 3D computational model of the sample, including surface concavities. Our system allowed for 3D model development of samples with a relative error of less than 1.2% and high repeatability in approximately three minutes. This was crucial for the extraction of the mechanical properties and subsequent inverse analysis, enabling the calibration of complex material models.

Mesenchymal stem cells injected into carotid artery to target focal brain injury home to perivascular space.

Rationale: The groundbreaking discovery of mesenchymal stem cells (MSCs) with their multifaceted benefits led to their widespread application in experimental medicine, including neurology. Efficient delivery of MSCs to damaged regions of the central nervous system may be a critical factor in determining outcome. Integrin VLA-4 (α4ß1) coded by ITGA4 and ITGB1 genes is an adhesion molecule expressed by leukocytes, which is responsible for initiation of their diapedesis through cell docking to the inflamed vessel wall expressing VCAM1 receptor. This function of VLA-4 has been recapitulated in neural stem cells and glial progenitors. Thus, it was prudent to investigate this tool as a vehicle driving extravasation of MSCs. Since MSCs naturally express ITGB1 subunit, we decided to supplement them with ITGA4 only. The purpose of our current study is to investigate the eventual fate of IA delivered ITGA4 engineered and naive MSCs. Methods: mRNA-ITGA4 transfected and naive MSCs were injected to right internal carotid artery of rats with focal brain injury. Through next three days MSC presence in animals' brain was navigated by magnetic resonance imaging. Transplanted cell location relative to the brain blood vessels and host immunological reaction were analyzed post-mortem by immunohistochemistry. The chemotaxis of modified and naive MSCs was additionally examined in in vitro transwell migration assay. Results: Both naïve and ITGA4-overexpressing cells remained inside the vascular lumen over the first two days after IA infusion. On the third day, 39% of mRNA-ITGA4 modified and 51% naïve MSCs homed to perivascular space in the injury region (p=NS). The gradual decrease of both naive and mRNA-ITGA4 transfected hBM-MSCs in the rat brain was observed. mRNA-ITGA4 transfected MSCs appeared to be more vulnerable to phagocytosis than naïve cells. Moreover, in vitro study revealed that homogenate from the injured brain repels migration of MSCs, corroborating the incomplete extravasation observed in vivo. Conclusions: In summary, IA transplanted MSCs are capable of homing to the perivascular space, an integral part of neurovascular unit, which might contribute to the replacement of injured pericytes, a critical element facilitating restoration of CNS function. The mRNA-ITGA4 transfection improves cell docking to vessel but this net benefit vanishes over the next two days due to fast clearance from cerebral vessels of the majority of transplanted cells, regardless of their engineering status. The drawbacks of mRNA-ITGA4 transfection become apparent on day 3 post transplantation due to the lower survival and higher vulnerability to host immune attack.

Immunomodulatory and Regenerative Effects of Mesenchymal Stem Cells and Extracellular Vesicles: Therapeutic Outlook for Inflammatory and Degenerative Diseases.

Mesenchymal stem cells (MSCs) are non-hematopoietic, multipotent stem cells derived from mesoderm, which can be easily isolated from many sources such as bone marrow, umbilical cord or adipose tissue. MSCs provide support for hematopoietic stem cells and have an ability to differentiate into multiple cell lines. Moreover, they have proangiogenic, protective and immunomodulatory properties. MSCs have the capacity to modulate both innate and adaptive immune responses, which accompany many diseases, by inhibiting pro-inflammatory reactions and stimulating anti-inflammatory activity. Recent findings revealed that the positive effect of MSCs is at least partly associated with the production of extracellular vesicles (EVs). EVs are small membrane structures, containing proteins, lipids and nuclei acids, which take part in intra-cellular communication. Many studies indicate that EVs contain protective and pro-regenerative properties and can modulate an immune response that is activated in various diseases such as CNS diseases, myocardial infarction, liver injury, lung diseases, ulcerative colitis or kidney injury. Thus, EVs have similar functions as their cells of origin and since they do not carry the risk of cell transplantation, such as tumor formation or small vessel blockage, they can be considered a potential therapeutic tool for cell-free therapy.

Human bone marrow mesenchymal stem cell-derived extracellular vesicles attenuate neuroinflammation evoked by focal brain injury in rats.

BACKGROUND: Ischemic stroke is the major cause of long-term severe disability and death in aged population. Cell death in the infarcted region of the brain induces immune reaction leading to further progression of tissue damage. Immunomodulatory function of mesenchymal stem cells (MSCs) has been shown in multiple preclinical studies; however, it has not been successfully translated to a routine clinical practice due to logistical, economical, regulatory, and intellectual property obstacles. It has been recently demonstrated that therapeutic effect of intravenously administered MSCs can be recapitulated by extracellular vesicles (EVs) derived from them. However, in contrast to MSCs, EVs were not capable to decrease stroke-induced neuroinflammation. Therefore, the aim of the study was to investigate if intra-arterial delivery of MSC-derived EVs will have stronger impact on focal brain injury-induced neuroinflammation, which mimics ischemic stroke, and how it compares to MSCs. METHODS: The studies were performed in adult male Wistar rats with focal brain injury induced by injection of 1 µl of 50 nmol ouabain into the right hemisphere. Two days after brain insult, 5 × 105 human bone marrow MSCs (hBM-MSCs) labeled with Molday ION or 1.3 × 109 EVs stained with PKH26 were intra-arterially injected into the right hemisphere under real-time MRI guidance. At days 1, 3, and 7 post-transplantation, the rats were decapitated, the brains were removed, and the presence of donor cells or EVs was analyzed. The cellular immune response in host brain was evaluated immunohistochemically, and humoral factors were measured by multiplex immunoassay. RESULTS: hBM-MSCs and EVs transplanted intra-arterially were observed in the rat ipsilateral hemisphere, near the ischemic region. Immunohistochemical analysis of brain tissue showed that injection of hBM-MSCs or EVs leads to the decrease of cell activation by ischemic injury, i.e., astrocytes, microglia, and infiltrating leucocytes, including T cytotoxic cells. Furthermore, we observed significant decrease of pro-inflammatory cytokines and chemokines after hBM-MSC or EV infusion comparing with non-treated rats with focal brain injury. CONCLUSIONS: Intra-arterially injected EVs attenuated neuroinflammation evoked by focal brain injury, which mimics ischemic stroke, and this effect was comparable to intra-arterial hBM-MSC transplantation. Thus, intra-arterial injection of EVs might be an attractive therapeutic approach, which obviates MSC-related obstacles.

The Biotic and Abiotic Carbon Monoxide Formation During Aerobic Co-digestion of Dairy Cattle Manure With Green Waste and Sawdust.

Carbon monoxide (CO), an air pollutant and a toxic gas to humans, can be generated during aerobic digestion of organic waste. CO is produced due to thermochemical processes, and also produced or consumed by cohorts of methanogenic, acetogenic, or sulfate-reducing bacteria. The exact mechanisms of biotic and abiotic formation of CO in aerobic digestion (particularly the effects of process temperature) are still not known. This study aimed to determine the temporal variation in CO concentrations during the aerobic digestion as a function of process temperature and activity of microorganisms. All experiments were conducted in controlled temperature reactors using homogeneous materials. The lab-scale tests with sterilized and non-sterilized mix of green waste, dairy cattle manure, sawdust (1:1:1 mass ratio) were carried out for 1 week at 10, 25, 30, 37, 40, 50, 60, 70°C to elucidate the biotic vs. abiotic effect. Gas concentrations of CO, O2, and CO2 inside the reactor were measured every 12 h. The CO concentrations observed for up to 30°C did not exceed 100 ppm v/v. For 50 and 60°C, significantly (p < 0.05) higher CO concentrations, reaching almost 600 ppm v/v, were observed. The regression analyses showed in both cases (sterile and non-sterile) a statistically significant effect (p < 0.05) of temperature on CO concentration, confirming that the increase in temperature causes an increase in CO concentration. The remaining factors (time, O2, and CO2 content) were not statistically significant (p > 0.05). A new polynomial model describing the effect of temperature, O2, and CO2 concentration on CO production during aerobic digestion of organic waste was formulated. It has been found that the proposed model for sterile variant had a better fit (R 2 = 0.86) compared with non-sterile (R 2 = 0.71). The model predicts CO emissions and could be considered for composting process optimization. The developed model could be further developed and useful for ambient air quality and occupational exposure to CO.

Oxytree Pruned Biomass Torrefaction: Process Kinetics.

Oxytree is a fast-growing energy crop with C4 photosynthesis. In this research, for the first time, the torrefaction kinetic parameters of pruned Oxytree biomass (Paulownia clon in Vitro 112) were determined. The influence of the Oxytree cultivation method and soil class on the kinetic parameters of the torrefaction was also investigated. Oxytree pruned biomass from a first-year plantation was subjected to torrefaction within temperature range from 200 to 300 °C and under anaerobic conditions in the laboratory-scale batch reactor. The mass loss was measured continuously during the process. The relative mass loss increased from 1.22% to 19.56% with the increase of the process temperature. The first-order constant rate reaction (k) values increased from 1.26 × 10-5 s-1 to 7.69 × 10-5 s-1 with the increase in temperature. The average activation energy for the pruned biomass of Oxytree torrefaction was 36.5 kJ∙mol-1. Statistical analysis showed no significant (p < 0.05) effect of the Oxytree cultivation method and soil class on the k value. The results of this research could be useful for the valorization of energy crops such as Oxytree and optimization of waste-to-carbon and waste-to-energy processes.

Neuroinflammation as a target for treatment of stroke using mesenchymal stem cells and extracellular vesicles.

Ischemic stroke is the third cause of death in the developed countries and the main reason of severe disability. Brain ischemia leads to the production of damage-associated molecular patterns (DAMPs) by neurons and glial cells which results in astrocyte and microglia activation, pro-inflammatory cytokines and chemokines production, blood-brain barrier (BBB) disruption, infiltration of leukocytes from the peripheral blood into the infarcted area, and further exacerbation of tissue damage. However, some immune cells such as microglia or monocytes are capable to change their phenotype to anti-inflammatory, produce anti-inflammatory cytokines, and protect injured nervous tissue. In this situation, therapies, which will modulate the immune response after brain ischemia, such as transplantation of mesenchymal stem cells (MSCs) are catching interest. Many experimental studies of ischemic stroke revealed that MSCs are able to modulate immune response and act neuroprotective, through stimulation of neurogenesis, oligodendrogenesis, astrogenesis, and angiogenesis. MSCs may also have an ability to replace injured cells, but the release of paracrine factors directly into the environment or via extracellular vesicles (EVs) seems to play the most pronounced role. EVs are membrane structures containing proteins, lipids, and nucleic acids, and they express similar properties as the cells from which they are derived. However, EVs have lower immunogenicity, do not express the risk of vessel blockage, and have the capacity to cross the blood-brain barrier. Experimental studies of ischemic stroke showed that EVs have immunomodulatory and neuroprotective properties; therefore, they can stimulate neurogenesis and angiogenesis. Up to now, 20 clinical trials with MSC transplantation into patients after stroke were performed, from which two concerned on only hemorrhagic stroke and 13 studied only on ischemic stroke. There is no clinical trial with EV injection into patients after brain ischemia so far, but the case with miR-124-enriched EVs administration is planned and probably there will be more clinical studies with EV transplantation in the near future.

Labeling of human mesenchymal stem cells with different classes of vital stains: robustness and toxicity.

BACKGROUND: Mesenchymal stem cell (MSC) transplantation has been explored as a new clinical approach to repair injured tissues. However, in order to evaluate the therapeutic activity of MSC, cell tracking techniques are required to determine the fate of transplanted cells in both preclinical and clinical studies. In these aspects, different vital stains offer the potential for labeling and monitoring of grafted cells in vivo. It is desirable to have tracking agents which have long-term stability, are not toxic to the cells, and do not affect cell function. METHODS: Here, we selected three different labels: CellTracker™ Green CMFDA, eGFP-mRNA (genetic pre-tag), and Molday ION Rhodamine B™ (nanoparticle-based fluorescent and magnetic label) and performed extensive analysis of their influence on in vitro expansion of human bone marrow-derived mesenchymal stem cells (hBM-MSCs), as well as potential of affecting therapeutic activity and the impact on the durability of staining. RESULTS: Our study showed that basic hBM-MSC characteristics and functions might be affected by labeling. We observed strong alterations of metabolic activity and morphology after eGFP and CellTracker™ Green CMFDA hBM-MSC staining. Molday ION Rhodamine B™ labeling revealed superior properties relatively to other vital stains. The relative expression level of most of the investigated growth factors remained stable after cell labeling, but we have observed some changes in the case of EGF, GDNF, HGF, and IGF gene expression. CONCLUSIONS: Taken together, we suggest performing similar to ours extensive analysis prior to using any cell label to tag MSC in experiments, as it can thoroughly bias results.