Constitutive activation of MET signaling impairs myogenic differentiation of rhabdomyosarcoma and promotes its development and progression

Rhabdomyosarcoma (RMS) is a soft tissue sarcoma, which may originate from impaired differentiation of mesenchymal stem cells (MSC). Expression of MET receptor is elevated in alveolar RMS subtype (ARMS) which is associated with worse prognosis, compared to embryonal RMS (ERMS). Forced differentiation of ARMS cells diminishes MET level and, as shown previously, MET silencing induces differentiation of ARMS. In ERMS cells introduction of TPR-MET oncogene leads to an uncontrolled overstimulation of the MET receptor downstream signaling pathways. In vivo, tumors formed by those cells in NOD-SCID mice display inhibited differentiation, enhanced proliferation, diminished apoptosis and increased infiltration of neutrophils. Consequently, tumors grow significantly faster and they display enhanced ability to metastasize to lungs and to vascularize due to elevated VEGF, MMP9 and miR-378 expression. In vitro, TPR-MET ERMS cells display enhanced migration, chemotaxis and invasion toward HGF and SDF-1. Introduction of TPR-MET into MSC increases survival and may induce expression of early myogenic factors depending on the genetic background, and it blocks terminal differentiation of skeletal myoblasts. To conclude, our results suggest that activation of MET signaling may cause defects in myogenic differentiation leading to rhabdomyosarcoma development and progression.     

Assessment of Effects of Laser Light Combining Three Wavelengths (450, 520 and 640 nm) on Temperature Increase and Depth of Tissue Lesions in an Ex Vivo Study

Background: Lasers are widely used in medicine in soft and hard tissue surgeries and biostimulation. Studies found in literature typically compare the effects of single-wavelength lasers on tissues or cell cultures. In our study, we used a diode laser capable of emitting three components of visible light (640 nm, red; 520 nm, green; 450 nm, blue) and combining them in a single beam. The aim of the study was to assess the effects of laser radiation in the visible spectrum on tissue in vitro, depending on the wavelength and pulse width. Methods: All irradiations were performed using the same output power (1.5 W). We used various duty cycles: 10, 50, 80 and 100% with 100 Hz frequency. Maximum superficial temperature, rate of temperature increase and lesion depth were investigated. Results: Maximum superficial temperature was observed for 450 + 520 nm irradiation (100% duty cycle). The highest rate of increase of temperature was noted for 450 + 520 nm (100% duty cycle). Maximum lesion depth was observed in case of three-wavelength irradiation (450 + 520 + 640 nm) for 100, 80 and 50% duty cycles. Conclusions: The synergistic effect of two-wavelength (450 + 520 nm) irradiation was observed in case of maximum temperature measurement. The deepest depth of lesion was noted after three-wavelength irradiation (450 + 520 + 640 nm).     

Thermal Insulation and Sound Absorption Properties of Open-Cell Polyurethane Foams Modified with Bio-Polyol Based on Used Cooking Oil

The main goal of this work was to evaluate the thermal insulation and sound absorption properties of open-cell rigid polyurethane foams synthesized with different contents of cooking oil-based polyol. The content of the applied bio-polyol as well as flame retardant (triethyl phosphate) in the foam formulation had a significant influence on the cellular structures of the materials. The open-cell polyurethane foams were characterized by apparent densities in the range 16–30 kg/m³. The sound absorption coefficients of the polyurethanes with various contents of bio-polyol were determined using the standing wave method (Kundt’s tube) in the frequency range of 100–6300 Hz. The effect of the content of the bio-polyol and flame retardant on the coefficient of thermal conductivity (at average temperatures of 0, 10 and 20 °C) as well as the compressive strength (at 20 and −10 °C) was analyzed. Different trends were observed in terms of the thermal insulation properties and sound absorption ability of the open-cell polyurethanes due to the addition of bio-polyol. In conclusion, it is necessary to use systems containing both petrochemical and bio-based raw materials.     

The Influence of Sub-Zero Conditions on the Mechanical Properties of Polylactide-Based Composites

Polylactide-based composites filled with waste fillers due to their sustainability are a subject of many current papers, in which their structural, mechanical, and thermal properties are evaluated. However, few studies focus on their behavior in low temperatures. In this paper, dynamic and quasi-static mechanical properties of polylactide-based composites filled with 10 wt% of linseed cake (a by-product of mechanical oil extraction from linseed) were evaluated at room temperature and at −40 °C by means of dynamic mechanical analysis (DMA), Charpy’s impact strength test and uniaxial tensile test. It was found that the effect of plasticization provided by the oil contained in the filler at room temperature is significantly reduced in sub-zero conditions due to solidification of the oil around −18 °C, as it was shown by differential scanning calorimetry (DSC) and DMA, but the overall mechanical performance of the polylactide-based composites was sufficient to enable their use in low-temperature applications.     

The reference liver – ABC and SLC drug transporters in healthy donor and metastatic livers

BackgroundAnalysis of results and conclusions in studies dedicated to pathology of the liver are usually based on comparison of pathological liver specimens and control/reference (considered as healthy) tissues. There are two main sources of the control liver samples used as the reference livers, i.e. deceased organ donor livers and non-tumorous tissue from metastatic livers, which are also applied for drug transporter investigations. However, no information has yet been published on drug transporters in these two major types of reference livers.MethodsWe explored ABC (P-gp, MRP1, MRP2, MRP3, MRP4, BCRP, BSEP) and SLC (NTCP, MCT1, OCT1, OCT3, OAT2, OATP1B1, OATP1B3, OATP2B1) family transporters expression (qPCR) and protein abundance (LC–MS/MS) in healthy donors (n = 9) and metastatic (n = 13) livers.ResultsThe analysis of mRNA content revealed significant differences in ABCB11, ABCC1, ABCG2, SLC10A1, SLC16A1, SLCO1B1 and SLCO2B1 gene expression between livers from organ donors and patients who underwent surgical resection of metastatic tumors. The protein abundance of NTCP was significantly higher, whereas of P-gp significantly lower in non-tumorous tissues from metastatic livers. Greater inter-individual variability in protein abundance of all studied transporters in subjects with metastatic colon cancer was also observed.ConclusionsThe results suggest that final conclusions in liver pathology studies may depend on the reference liver tissue used, especially in gene expression studies.     

Digital movie analysis for quantification of beating frequencies, chronotropic effects, and beating areas in cardiomyocyte cultures

Software, named Cardio Analyser, was developed for digital movie analysis of beating frequencies, drug-induced chronotropic effects, and quantification of beating areas of contracting cardiomyocyte cultures. A major novelty of the software is the introduction of automated noise filtering and automated movie analysis of beating frequencies and areas of contracting cardiomyocyte cultures. The software was based on the observation that the intensity of light transmitted through a contractive tissue changes periodically in a way that correlates with the contractions. We provided proof of principle for the method by derivation of relevant data from movies of multicellular cardiomyocyte cultures derived from embryonic stem cells. Moreover, we compared the data to equivalent results obtained by extracellular electric field potential recordings. The comparison demonstrated higher sensitivity to chronotropic effects of the beta-adrenoceptor agonist isoprenaline, and hence implied that more embryonic stem cells underwent differentiation into beta-adrenoceptor-responding cardiomyocytes, in the experimental setup applied for movie analysis than in the setup used for extracellular electric field potential recordings. Our study indicates that the movie analysis method may have potential to be optimized for screening in early drug discovery, aiming to identify cardiac drug candidates or to alert for adverse effects on heart functionality or embryonic heart development.     

Discovery of 5-[[4-[(2,3-dimethyl-2H-indazol-6-yl)methylamino]-2-pyrimidinyl]amino]-2-methyl-benzenesulfonamide (Pazopanib), a novel and potent vascular endothelial growth factor receptor inhibitor

Inhibition of the vascular endothelial growth factor (VEGF) signaling pathway has emerged as one of the most promising new approaches for cancer therapy. We describe herein the key steps starting from an initial screening hit leading to the discovery of pazopanib, N(4)-(2,3-dimethyl-2H-indazol-6-yl)-N(4)-methyl-N(2)-(4-methyl-3-sulfonamidophenyl)-2,4-pyrimidinediamine, a potent pan-VEGF receptor (VEGFR) inhibitor under clinical development for renal-cell cancer and other solid tumors.