A Cellular Model of Amyotrophic Lateral Sclerosis to Study the Therapeutic Effects of Extracellular Vesicles from Adipose Mesenchymal Stem Cells on Microglial Activation.

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by the progressive degeneration of upper and lower motor neurons (MNs) in the brain and spinal cord, leading to progressive paralysis and death. Increasing evidence indicates that neuroinflammation plays an important role in ALS's pathogenesis and disease progression. Neuroinflammatory responses, primarily driven by activated microglia and astrocytes, and followed by infiltrating peripheral immune cells, contribute to exacerbate/accelerate MN death. In particular, the role of the microglia in ALS remains unclear, partly due to the lack of experimental models that can fully recapitulate the complexity of ALS's pathology. In this study, we developed and characterized a microglial cell line, SIM-A9-expressing human mutant protein Cu+/Zn+ superoxide dismutase_1 (SIM-A9hSOD1(G93A)), as a suitable model in vitro mimicking the microglia activity in ALS. The expression of hSOD1(G93A) in SIM-A9 cells induced a change in their metabolic activity, causing polarization into a pro-inflammatory phenotype and enhancing reactive oxygen species production, which is known to activate cell death processes and apoptosis. Afterward, we used our microglial model as an experimental set-up to investigate the therapeutic action of extracellular vesicles isolated from adipose mesenchymal stem cells (ASC-EVs). ASC-EVs represent a promising therapeutic treatment for ALS due to their neuroprotective and immunomodulatory properties. Here, we demonstrated that treatment with ASC-EVs is able to modulate activated ALS microglia, reducing their metabolic activity and polarizing their phenotype toward an anti-inflammatory one through a mechanism of reduction of reactive oxygen species.

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.

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.

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.

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.

A Breakthrough Application of a Cross-Linked Polystyrene Anion-Exchange Membrane for a Hydrogencarbonate Ion-Selective Electrode.

Polystyrene cross-linked with divinylbenzene and functionalized by a quaternary ammonium cation anion site is used as the membrane of a hydrogencarbonate (i.e., bicarbonate) ion-selective electrode. The polystyrene matrix membrane improves the selectivity towards interfering lipophilic ions in comparison to previously described polyvinyl chloride membranes. The reason for this behaviour is sought in coupled ion-exchange and pore-diffusion processes in the membrane and the resulting kinetic discrimination of interfering ions. The electrode is successfully used for determination of bicarbonates in mineral drinking waters. The simplex method is employed to refine the analytical outcome.

Adipose mesenchymal stem cells-derived extracellular vesicles exert their preferential action in damaged central sites of SOD1 mice rather than peripherally.

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder involving motor neuron (MN) loss in the motor cortex, brainstem and spinal cord leading to progressive paralysis and death. Due to the pathogenetic complexity, there are no effective therapies available. In this context the use of mesenchymal stem cells and their vesicular counterpart is an emerging therapeutic strategy to counteract neurodegeneration. The extracellular vesicles derived from adipose stem cells (ASC-EVs) recapitulate and ameliorate the neuroprotective effect of stem cells and, thanks to their small dimensions, makes their use suitable to develop novel therapeutic approaches for neurodegenerative diseases as ALS. Here we investigate a therapeutic regimen of ASC-EVs injection in SOD1(G93A) mice, the most widely used murine model of ALS. Repeated intranasal administrations of high doses of ASC-EVs were able to ameliorate motor performance of injected SOD1(G93A) mice at the early stage of the disease and produce a significant improvement at the end-stage in the lumbar MNs rescue. Moreover, ASC-EVs preserve the structure of neuromuscular junction without counteracting the muscle atrophy. The results indicate that the intranasal ASC-EVs administration acts in central nervous system sites rather than at peripheral level in SOD1(G93A) mice. These considerations allow us to identify future applications of ASC-EVs that involve different targets simultaneously to maximize the clinical and neuropathological outcomes in ALS in vivo models.

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.

The Impact of Abiotic and Biotic Conditions for Degradation Behaviors of Common Biodegradable Products in Stabilized Composts.

This work examines the influence of the degradation behaviors of biotic and abiotic conditions on three types of biodegradable products: cups from PLA and from cellulose, and plates from sugarcane. The main objective of this study was to evaluate if biodegradable products can be degraded in composts that were stabilized by backyard composting. Furthermore, the impact of crucial abiotic parameters (temperature and pH) for the degradation behaviors process was investigated. The changes in the biopolymers were analyzed by FTIR spectroscopy. This work confirmed that abiotic and biotic conditions are important for an effective disintegration of the investigated biodegradable products. Under abiotic conditions, the degradation behaviors of PLA were observable under both tested temperature (38 and 59 °C) conditions, but only at the higher temperature was complete disintegration observed after 6 weeks of incubation in mature compost. Moreover, our research shows that some biodegradable products made from cellulose also need additional attention, especially with respect to incorporated additives, as composting could be altered and optimal conditions in composting may not be achieved. This study shows that the disintegration of biodegradable products is a comprehensive process and requires detailed evaluation during composting. The results also showed that biodegradable products can also be degraded post composting and that microplastic pollution from biodegradable polymers in soil may be removed by simple physical treatments.

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.

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.

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.

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.

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.

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.

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.

Computed tomography as a source of electron density information for radiation treatment planning.

PURPOSE: To evaluate the performance of computed tomography (CT) systems of various designs as a source of electron density (rho(el)) data for treatment planning of radiation therapy. MATERIAL AND METHODS: Dependence of CT numbers on relative electron density of tissue-equivalent materials (HU-rho(el) relationship) was measured for several general-purpose CT systems (single-slice, multislice, wide-bore multislice), for radiotherapy simulators with a single-slice CT and kV CBCT (cone-beam CT) options, as well as for linear accelerators with kV and MV CBCT systems. Electron density phantoms of four sizes were used. Measurement data were compared with the standard HU-rhoel relationships predefined in two commercial treatment-planning systems (TPS). RESULTS: The HU-rho(el) relationships obtained with all of the general-purpose CT scanners operating at voltages close to 120 kV were very similar to each other and close to those predefined in TPS. Some dependency of HU values on tube voltage was observed for bone- equivalent materials. For a given tube voltage, differences in results obtained for different phantoms were larger than those obtained for different CT scanners. For radiotherapy simulators and for kV CBCT systems, the information on rhoel was much less precise because of poor uniformity of images. For MV CBCT, the results were significantly different than for kV systems due to the differing energy spectrum of the beam. CONCLUSION: The HU-rho(el) relationships predefined in TPS can be used for general-purpose CT systems operating at voltages close to 120 kV. For nontypical imaging systems (e.g., CBCT), the relationship can be significantly different and, therefore, it should always be measured and carefully analyzed before using CT data for treatment planning.