Cellular Response to Individual Components of the Platelet Concentrate

Platelet concentrates and especially their further product platelet lysate, are widely used as a replacement for cell culturing. Platelets contain a broad spectrum of growth factors and bioactive molecules that affect cellular fate. However, the cellular response to individual components of the human platelet concentrate is still unclear. The aim of this study was to observe cellular behavior according to the individual components of platelet concentrates. The bioactive molecule content was determined. The cells were supplemented with a medium containing 8% (v/v) of platelet proteins in plasma, pure platelet proteins in deionized water, and pure plasma. The results showed a higher concentration of fibrinogen, albumin, insulin growth factor I (IGF-1), keratinocyte growth factor (KGF), and hepatocyte growth factor (HGF), in the groups containing plasma. On the other hand, chemokine RANTES and platelet-derived growth factor bb (PDGF-bb), were higher in the groups containing platelet proteins. The groups containing both plasma and plasma proteins showed the most pronounced proliferation and viability of mesenchymal stem cells and fibroblasts. The platelet proteins alone were not sufficient to provide optimal cell growth and viability. A synergic effect of platelet proteins and plasma was observed. The data indicated the importance of plasma in platelet lysate for cell growth.     

Causes of Decreased Discharge and Damage to a Dewatering Well’s Gravel Coat

Drainage wells used in lignite mining have to be operated at high efficiency in order to achieve a targeted drawdown in the shortest time period possible. However, oversizing of pump units and excessive pumping can cause accelerated aging of wells and over time, decrease a well’s drainage efficiency. This article analyzes the case of a well that was operated for 10 years and then abandoned at a lignite mine. A common practice in installing dewatering wells in Poland is to protect the well screen with a pre-fabricated granular layer (called a “gravel coat”) that is attached to the screen when it is lowered into the hole. The combination of the gravel coat and well screen is surrounded by a protective gravel pack. Samples of the gravel coat were obtained from the abandoned well. The percentage by weight of precipitates in, and the porosity of the gravel coat from the abandoned well were determined and compared with the properties of new gravel coat material. Tests were conducted in the laboratory to compare hydraulic losses, apparent seepage velocity, effective velocity, and the hydraulic efficiency of the well’s gravel coat. The results of the tests explain the low hydraulic efficiency of the abandoned well and the origin of the damage to the gravel coat. The actual yield was also compared with the calculated theoretical safe yield, which was estimated based on the results of geological records from the construction of the well.     

Occurrence of estrogenic mycotoxin - Zearalenone in aqueous environmental samples with various NOM content

Zearalenone is a mycotoxin produced by some Fusarium species in food and feed. Toxicity of ZEA and its metabolites is related to the chemical structure of the mycotoxins, similar to naturally occurring estrogens. Currently, there is increasing awareness of the significance of fungi and their toxic metabolites in water. Thus it was considered essential to determine the concentration of estrogenic toxin in surface waters, groundwater and wastewater in Poland. The application of ZearalaTest immunoaffinity columns for zearalenone determination by HPLC method in water samples characterized by different natural organic matter (NOM) content gave the recovery rate ranging from 74 to 86%. Obtained results showed that the natural organic micropollutants of water with low molecular masses have an effect on zearalenone recovery. Zearalenone was present in water samples in the range from 0 to 43.7 ng L⁻¹.     

More Than Meets the Eye Regarding Cancer Metabolism

In spite of the continuous improvement in our knowledge of the nature of cancer, the causes of its formation and the development of new treatment methods, our knowledge is still incomplete. A key issue is the difference in metabolism between normal and cancer cells. The features that distinguish cancer cells from normal cells are the increased proliferation and abnormal differentiation and maturation of these cells, which are due to regulatory changes in the emerging tumour. Normal cells use oxidative phosphorylation (OXPHOS) in the mitochondrion as a major source of energy during division. During OXPHOS, there are 36 ATP molecules produced from one molecule of glucose, in contrast to glycolysis which provides an ATP supply of only two molecules. Although aerobic glucose metabolism is more efficient, metabolism based on intensive glycolysis provides intermediate metabolites necessary for the synthesis of nucleic acids, proteins and lipids, which are in constant high demand due to the intense cell division in cancer. This is the main reason why the cancer cell does not “give up” on glycolysis despite the high demand for energy in the form of ATP. One of the evolving trends in the development of anti-cancer therapies is to exploit differences in the metabolism of normal cells and cancer cells. Currently constructed therapies, based on cell metabolism, focus on the attempt to reprogram the metabolic pathways of the cell in such a manner that it becomes possible to stop unrestrained proliferation.     

Liquid Biopsy in Melanoma: Significance in Diagnostics,Prediction and Treatment Monitoring

Liquid biopsy is a common term referring to circulating tumor cells and other biomarkers, such as circulating tumor DNA (ctDNA) or extracellular vesicles. Liquid biopsy presents a range of clinical advantages, such as the low invasiveness of the blood sample collection and continuous control of the tumor progression. In addition, this approach enables the mechanisms of drug resistance to be determined in various methods of cancer treatment, including immunotherapy. However, in the case of melanoma, the application of liquid biopsy in patient stratification and therapy needs further investigation. This review attempts to collect all of the relevant and recent information about circulating melanoma cells (CMCs) related to the context of malignant melanoma and immunotherapy. Furthermore, the biology of liquid biopsy analytes, including CMCs, ctDNA, mRNA and exosomes, as well as techniques for their detection and isolation, are also described. The available data support the notion that thoughtful selection of biomarkers and technologies for their detection can contribute to the development of precision medicine by increasing the efficacy of cancer diagnostics and treatment.     

Synaptic Alterations in a Transgenic Model of Tuberous Sclerosis Complex: Relevance to Autism Spectrum Disorders

Tuberous sclerosis complex (TSC) is a rare, multi-system genetic disease with serious neurological and mental symptoms, including autism. Mutations in the TSC1/TSC2 genes lead to the overactivation of mTOR signalling, which is also linked to nonsyndromic autism. Our aim was to analyse synaptic pathology in a transgenic model of TSC: two-month-old male B6;129S4-Tsc2^tm1Djk/J mice with Tsc2 haploinsufficiency. Significant brain-region-dependent alterations in the expression of several synaptic proteins were identified. The most prominent changes were observed in the immunoreactivity of presynaptic VAMP1/2 (ca. 50% increase) and phospho-synapsin-1 (Ser62/67) (ca. 80% increase). Transmission electron microscopy demonstrated serious ultrastructural abnormalities in synapses such as a blurred structure of synaptic density and a significantly increased number of synaptic vesicles. The impairment of synaptic mitochondrial ultrastructure was represented by excessive elongation, swelling, and blurred crista contours. Polyribosomes in the cytoplasm and swollen Golgi apparatus suggest possible impairment of protein metabolism. Moreover, the delamination of myelin and the presence of vacuolar structures in the cell nucleus were observed. We also report that Tsc2^+/− mice displayed increased brain weights and sizes. The behavioural analysis demonstrated the impairment of memory function, as established in the novel object recognition test. To summarise, our data indicate serious synaptic impairment in the brains of male Tsc2^+/− mice.     

Free-ranging European bisons accumulate more cadmium in the liver and kidneys than domestic cattle in north-eastern Poland

It has been shown that free-ranging big game animals accumulate several-fold more cadmium (Cd) in the liver and kidneys than domestic animals. To examine possible reasons for this difference, in the present work we determined the concentrations of Cd in the liver and kidney cortex of European bisons (n=23) from Bia?owieza Forest (north-eastern Poland) and domestic cattle (n=15) from the same region; in addition, analyses of Cd in the grasses and soil as well as of soil pH were carried out. The accumulation of Cd in liver and kidney cortex of the female bisons correlated significantly with the age up to 7 years, but stabilized thereafter. The 7-12-year-old bisons had 2.14- and 2.25-fold higher concentrations of Cd in the liver and kidney cortex, respectively, than the age-matched domestic cattle. Notably, the Cd levels in the liver and kidneys of the 8-12-year-old cattle were comparable to those found in the 2-year-old and 4-6-year-old bisons, respectively. The content of Cd in the grasses from Bia?owieza Forest appeared to be 2.1-fold higher than that in the plants from the pastures. Similarly, the concentration of water-extractable Cd in the soil was 2.7-fold greater in Bia?owieza Forest than in the pastures, despite the fact that nitric acid-extractable Cd (total Cd) was similar in the soils from the two sites. The concentration of water-extractable Cd in the soil as well as the content of Cd in the grasses inversely correlated with soil pH, which appeared to be significantly lower in Bia?owieza Forest. These data indicate that soil pH is probably responsible for the higher concentrations of Cd in the feed and tissues of bisons as compared with those of domestic cattle.     

A new insight into in vitro behaviour of poly(ε-caprolactone)/bioactive glass composites in biologically related fluids

In the present work, the role of content, size and chemical composition of gel-derived bioactive glass particles from the SiO₂–CaO–P₂O₅ system in modulating the in vitro bioactivity, osteoinductive properties and long-term (up to 15 months) degradation behaviour of poly(ε-caprolactone)-based composite films was investigated. Bioactivity was assessed in simulated body fluid (SBF) and HEPES-free Dulbecco’s modified Eagle medium (DMEM) supplemented with 10% foetal bovine serum (FBS), while hydrolytic degradation tests were performed in phosphate buffer saline. Obtained composite films showed excellent calcium phosphate (CaP) layer forming ability in both SBF and DMEM-10% FBS. However, kinetics of bioactivity process strongly depended on the type of medium used. The layer of amino acids and proteins, derived from cell culture medium, on the surfaces of composites created barrier that inhibited release of the ions on the one hand, while increasing nucleation density of calcium phosphates, affecting the morphology of formed CaP layers on the other. The presence of bioactive glass fillers was shown to impart osteoinductive properties to obtained films, supporting osteoblast attachment and proliferation, as well as stimulating cell differentiation and also matrix mineralization process in vitro. We showed that kinetics of bioactivity process and also osteoinductive properties of composite films could be easily modulated with the use of different contents and chemical compositions of fillers. The results showed that modification of PCL matrix with bioactive glass particles accelerated its degradation. We proved that the degradation rate of composites could be controlled and optimized for bone regeneration, in particular by using bioactive fillers causing different calcium phosphate layer forming ability on the surfaces of composites, depending on particle size and chemical composition. We have presented new opportunities to design and obtain multifunctional composites with tunable degradation and bioactivity kinetics, as well as biological properties that can meet complex requirements of bone tissue engineering.     

Feasibility of diffusion-weighted imaging with DWIBS in staging Hodgkin lymphoma in pediatric patients: comparison with PET/CT

Objective

The aim of the study was to evaluate feasibility of diffusion-weighted whole-body imaging with background body signal suppression (DWIBS) method in diagnosing Hodgkin lymphoma in pediatric patients and to compare it with 18F-FDG PET/CT as a gold standard.

Materials and methods

Eleven patients (median age 14) with newly diagnosed Hodgkin lymphoma were examined with 18F-FDG PET/CT and MRI including whole-body DWIBS sequence (b = 0, 800 s/mm²), before the oncologic treatment. About 26 locations of lymphatic tissues were evaluated visually and quantitatively using ADC_mean (DWIBS) and SUV_max (18F-FDG PET/CT), respectively.

Results

All affected lymph node regions (n = 134) diagnosed in 18F-FDG PET/CT were found with DWIBS, presenting decreased diffusion. Significant correlation was found between ADC and SUV values (R² = − 0.37; p = 0.0001). Nevertheless, additional 33 regions were recognized only by DWIBS. They were significantly smaller than regions diagnosed by both methods.

Discussion

Agreement between DWIBS and 18F-FDG PET/CT for detection and staging of malignant lymphoma is high. DWIBS can be used for the evaluation of pediatric Hodgkin lymphoma.