Petrographical and organic geochemical study of the lignite from the Smederevsko Pomoravlje field (Kostolac Basin, Serbia)

Three Upper Miocene (Pontian) lignite seams are present in the Smederevsko Pomoravlje field (Kostolac Basin, Serbia). The origin of their organic matter (OM), the characteristics of the depositional environment and certain utilisation properties have been evaluated based on petrographic data, bulk organic geochemical parameters, biomarker patterns and their isotope signatures. Moreover, results of isotopic analysis were used for the investigation of the influence of diagenetic aromatisation on delta C-13 signatures of biomarkers. The studied lignites are typical humic coals. The OM of lignites is derived from woody vegetation and herbaceous peat-forming plants, with a strong prevalence of the former. The peat-forming vegetation is dominated by decay resistant conifers, including gymnosperm families Cupressaceae, Taxodiaceae, and Pinaceae. Angiosperms occurred in lower amounts. Minor contribution of ferns, fungi and emergent aquatic macrophyta to the biomass is also evident. Chemoautotrophic- and heterotrophic bacteria played an import role during diagenesis. Diagenetic alterations, associated with change in the number of carbon atoms, influence delta C-13 ratios. Diagenetic aromatisation of di- and non-hopanoid triterpenoids is accompanied with C-13 depletion, whereas aromatisation of hopanoids displays the opposite trend. Peatification proceeded in a fresh water environment under variable, anoxic to slightly oxic redox conditions. The lowermost coal seam III accumulated in a topogenous fresh water peat mire with open water areas, which changed occasionally into a wet forest swamp. This resulted in the deposition of mineral-rich coal. The characteristics of lignite in coal seam II are similar to those of coal seam III. This is supported also by generally similar delta C-13 values of individual biomarkers. Coal seam I is dominated by xylite-rich coal, formed under mesotrophic to ombrotrophic conditions. Rapid flooding of the bogs stopped peat growth in all three coal seams. The ratios of ring-A-degraded and non-degraded aromatic diterpenoids and non-hopanoid triterpenoids, proposed in this study, as well as degree of aromatisation of these biomarkers, reflect changes in the water table. Calorific values of the samples indicate that they meet basic requirements for utilisation in the thermal power plants. None of the lignite samples is suitable for coal briquetting, whereas, based on petrographic data, lignite from coal seam I possesses certain potential for fluidized bed gasification

Hybridosomes from spruce needle homogenate

Introduction: Being of compatible structure with biomembranes, lipid–based nanoparticles are considered as convenient platforms for drug delivery systems. In the proposed work we considered formation of lipid nanovesicles associated with bioactive phytochemicals from spruce needle homogenate (here called hybridosomes). We formed hybridosomes by mixing appropriate amounts of lecithin, supernatant of isolation of extracellular particles from spruce needle homogenate and glycerol. Methods: We visualized hybridosomes by light microscopy and cryogenic transmission electron microscopy and assessed them by flow cytometry, dynamic light scattering, ultraviolet–visual spectroscopy and interferometric microscopy. Results: We found that the particles consisted of a bilayer membrane and a fluid-like interior. Flow cytometry and interferometric light microscopy measurements showed that the majority of the particles were nano-sized. Dynamic light scattering and interferometric light microscopy measurements agreed well with the determined average hydrodynamic radius of the particles Rh (between 140 and 180 nm) while their number densities were in the range between 10^13 and 10^14/mL indicating that hybridosomes present about 2/3 of the mixture, excluding solvent and other small molecules. Discussion: Simple and low-cost preparation method, non-demanding saving process and efficient formation procedure suggest that large scale production of hybridosomes from lipids and spruce needle homogenate is feasible.Small New World 2.0 4-5 September 2023., Graz, Austri

Hybridosomes from spruce needle homogenate

Introduction: Being of compatible structure with biomembranes, lipid–based nanoparticles are considered as convenient platforms for drug delivery systems. In the proposed work we considered formation of lipid nanovesicles associated with bioactive phytochemicals from spruce needle homogenate (here called hybridosomes). We formed hybridosomes by mixing appropriate amounts of lecithin, supernatant of isolation of extracellular particles from spruce needle homogenate and glycerol. Methods: We visualized hybridosomes by light microscopy and cryogenic transmission electron microscopy and assessed them by flow cytometry, dynamic light scattering, ultraviolet–visual spectroscopy and interferometric microscopy. Results: We found that the particles consisted of a bilayer membrane and a fluid-like interior. Flow cytometry and interferometric light microscopy measurements showed that the majority of the particles were nano-sized. Dynamic light scattering and interferometric light microscopy measurements agreed well with the determined average hydrodynamic radius of the particles Rh (between 140 and 180 nm) while their number densities were in the range between 10^13 and 10^14/mL indicating that hybridosomes present about 2/3 of the mixture, excluding solvent and other small molecules. Discussion: Simple and low-cost preparation method, non-demanding saving process and efficient formation procedure suggest that large scale production of hybridosomes from lipids and spruce needle homogenate is feasible.Small New World 2.0 4-5 September 2023., Graz, Austri

Characterization of Nanohybridosomes from Lipids and Spruce Homogenate Containing Extracellular Vesicles

Introduction: Lipid nanovesicles associated with bioactive phytochemicals from spruce needle homogenate (here called nano-sized hybridosomes or nanohybridosomes, NSHs) were considered. Methods: We formed NSHs by mixing appropriate amounts of lecithin, glycerol and supernatant of isolation of extracellular vesicles from spruce needle homogenate. We visualized NSHs by light microscopy and cryogenic transmission electron microscopy and assessed them by flow cytometry, dynamic light scattering, ultraviolet–visual spectroscopy, interferometric light microscopy and liquid chromatography–mass spectrometry. Results: We found that the particles consisted of a bilayer membrane and a fluid-like interior. Flow cytometry and interferometric light microscopy measurements showed that the majority of the particles were nano-sized. Dynamic light scattering and interferometric light microscopy measurements agreed well on the average hydrodynamic radius of the particles Rh (between 140 and 180 nm), while the concentrations of the particles were in the range between 1013 and 1014/mL indicating that NSHs present a considerable (more than 25%) of the sample which is much more than the yield of natural extracellular vesicles (EVs) from spruce needle homogenate (estimated less than 1%). Spruce specific lipids and proteins were found in hybridosomes. Discussion: Simple and low-cost preparation method, non-demanding saving process and efficient formation procedure suggest that large-scale production of NSHs from lipids and spruce needle homogenate is feasible. Plain Language Summary: Cells shed into their exterior nanoparticles (here referred to as extracellular vesicles – EVs) that are free to move, reach distant cells and are taken up by them. As they carry bioactive constituents, EVs may have important impact on the recipient cells. The mechanisms of EV formation and mediation can be employed in designing therapeutic, prophylactic and diagnostic methods for various medical issues. EVs can be harvested from biological samples; however, their yield is small,12 and there are potential side effects. Artificial vesicles – liposomes – have high yield; however, in vivo, they can be degraded before reaching the target and their reproducibility is yet insufficient. In order to combine advantages of both types of nanoparticles, we have composed nanohybridosomes (NSHs) from soya lecithin, water and supernatant of isolation of EVs from spruce needle homogenate, visualized them by cryogenic electron microscopy and characterized them with respect to their size, concentration and protein/nucleic acid content. We have applied a recently developed interferometric light microscopy to determine the hydrodynamic radius and the concentration of EVs. We found that the majority of composed particles are nano-sized and that they enclose more than 25% of the incoming volume of liquid, which is considerably more than about 1% that can be harvested by isolation of EVs from spruce needle homogenate by (ultra) centrifugatio

Polymorphic expression of glutathione transferases A1, M1, P1 and T1 in epithelial ovarian cancer: a Serbian case-control study

PURPOSE: Since several studies have proposed that epithelial ovarian cancer should not be considered as a single disease entity and that it results from an accumulation of genetic changes, we aimed to assess the polymorphic expression of major cytosolic glutathione S-transferases (GSTM1, T1, A1 and P1) with respect to ovarian cancer susceptibility and aggressiveness. METHODS: This case-control study was conducted on 93 newly diagnosed epithelial ovarian cancer patients and 178 healthy matched controls. The multiplex polymerase chain reaction (PCR) was used to detect homozygous deletions of GSTM1 and GSTT1 genes. Analysis of the single nucleotide polymorphism (SNP) GSTA1 C69T was performed using PCR-restriction fragment length polymorphism (RFLP), while for SNP GSTP1 Ile105Val real-time PCR was used. RESULTS: No significant association to ovarian cancer risk was found for individual GSTM1, GSTA1 and GSTP1 genotypes (p>0.05). However, the carriers of GSTT1-active genotype were at 2-fold higher risk of ovarian cancer development (95%CI: 1.00-4.01, p=0.049), which was even more elevated in the subgroup of patients with positive family history of cancer. Moreover, the frequency of all three GST genotypes that might be associated to ovarian cancer risk (GSTT1-active, GSTA1-active and GSTP1-referent) was significantly higher in patients than in the control group (p=0.042). Even more, patients who were carriers of combination of these three genotypes represented over 64% of the total number of patients within any of the International Federation of Gynecology and Obstetrics (FIGO) stages of ovarian cancer. CONCLUSIONS: This study provides supportive evidence that GSTs might affect both susceptibility and progression of ovarian cance

BMP axis in cancer cachexia

BACKGROUND Cancer cachexia is a devastating metabolic syndrome characterized by systemic inflammation and massive muscle and adipose tissue wasting. Although cancer cachexia is responsible for about 25% of cancer deaths, no effective therapies are available, and the underlying mechanisms have not been fully elucidated. Its occurrence complicates patients’ management, reduces tolerance to treatments and negatively affects patient quality of life. Muscle wasting, mainly due to increased protein breakdown rates, is one of the most prominent features of cachexia. Blocking muscle loss in cachexia mouse models dramatically prolongs survival even of animals in which tumor growth is not inhibited. Recent observations showed that bone morphogenetic protein (BMP) signaling, acting through Smad1, Smad5 and Smad8 (Smad1/5/8), is a master regulator of muscle homeostasis. BMP-Smad1/5/8 axis negatively regulates a novel ubiquitin ligase (MUSA1) required for muscle loss induced by denervation. MATERIALS AND METHODS First aim of the present work was to test if alterations of the BMP signaling pathway occur in cancer-induced muscle wasting in patients. For this purpose we checked the state of activation of the BMP pathway in muscle of cachectic vs non–cachectic patients affected by colon, pancreatic and esophagus cancer and in control subjects. We checked by Western Blot the phosphorylation levels of Smad1/5/8 and of Smad3 and by quantitative Real-Time PCR (qRT-PCR) the expression levels of different atrophy-related genes The second aim was to evaluate the degree of muscle atrophy and distribution of muscle fibers in patients and control subjects using morphometric and immunohistochemical analyses. We also performed analysis on distribution of NCAM positive muscle fibers to assess the effect of denervation on muscle tropism. RESULTS From December 2014 we collected 95 rectus abdominis muscle biopsies of cancer patients and 11 from control subjects. In line with the results we obtained in C26 mice model (a well-established cancer cachexia experimental model) Smad1/5/8 phosphorylation, readout of the state of activation of the BMP pathway, was nearly completely abrogated in the muscles of cancer cachectic patients compared to cancer non-cachectic ones. Interestingly, the level of phosphorylation of Smad3 was not significantly affected suggesting specific effects of cancer growth on BMP pathway. The expression levels of different atrophy-related genes including MUSA1 were induced in the cachectic muscles. Interestingly, several BMP related genes are also changing the expression during cancer growth. We also found a correlation between suppression of BMP pathway, expression of atrophy related genes and Noggin, known to block BMP pathway. Morphometric analysis shown that patients with cancer cachexia have smaller myofiber diameter (in particular fast type fibers) in comparison to age-matched controls. In skeletal muscle from cancer patients (either cachectic or non-cachectic) we detected a prevalence of flat shaped, angulated and severely atrophic myofibers (i.e. morphological features of denervated myofibers), big fiber-type grouping (i.e. typical hallmark of denervation/reinnervation events) and numerous NCAM positive myofibers (i.e. specific marker of denervation). CONCLUSIONS These findings are consistent with the hypothesis that BMP inhibition is permissive to cachexia onset. Since the reactivation of the BMP-dependent signaling and MUSA1 suppression was sufficient to prevent tumor-induced muscle atrophy in our C26 mouse model (data not shown), the present data suggest that the BMP axis can be an effective target for therapeutic approaches to counteract cachexia also in cancer patients. The results of morphometric and immunohistochemical studies collected till now may suggest that denervation contributes to myofiber atrophy in cancer cachexia

Model of acceptance and use of market-ready home-based e-care services

Because of population aging, home-based e-care services (HBECSs) have raised interest among users and service providers. Recently, scholars have focused extensively on the needs and motives of older adults as care receivers that shape their pre-implementation acceptance of such technologies. Yet, little is known to date about post-implementation experiences and interrelationships between acceptance factors of market-ready services among care receivers and caregivers. To fill this research gap, an intervention study lasting up to eight weeks tested three market-ready HBECSs. Semi-structured interviews were conducted with seven informal caregivers and six care receivers. Qualitative analysis combining grounded theory with thematic analysis was used to present a thematic description of participants’ experiences and inductively develop a substantive model of HBECS acceptance and use. The results detail the impact and expected benefits of such technologies and various barriers to HBECSs use in conjunction with their functionalities and users’ social interactions. Acceptance and future use are determined by a complex mix of interrelated factors. These range from contextual circumstances to characteristics of the caregivers and care receivers to the service properties and perceived outcomes of use, such as safety, psychological relief, and peace of mind