Human Umbilical Vein Endothelial Cells as a Versatile Cellular Model System in Diverse Experimental Paradigms: An Ultrastructural Perspective.

Human umbilical vein endothelial cells (HUVECs) are primary cells isolated from the vein of an umbilical cord, extensively used in cardiovascular studies and medical research. These cells, retaining the characteristics of endothelial cells in vivo, serve as a valuable cellular model system for understanding vascular biology, endothelial dysfunction, pathophysiology of diseases such as atherosclerosis, and responses to different drugs or treatments. Transmission electron microscopy (TEM) has been a cornerstone in revealing the detailed architecture of multiple cellular model systems including HUVECs, allowing researchers to visualize subcellular organelles, membrane structures, and cytoskeletal elements. Among them, the endoplasmic reticulum, Golgi apparatus, mitochondria, and nucleus can be meticulously examined to recognize alterations indicative of cellular responses to various stimuli. Importantly, Weibel-Palade bodies are characteristic secretory organelles found in HUVECs, which can be easily distinguished in the TEM. These distinctive structures also dynamically react to different factors through regulated exocytosis, resulting in complete or selective release of their contents. This detailed review summarizes the ultrastructural features of HUVECs and highlights the utility of TEM as a pivotal tool for analyzing HUVECs in diverse research frameworks, contributing valuable insights into the comprehension of HUVEC behavior and enriching our knowledge into the complexity of vascular biology.

Impacts of iron on ultrastructural features of NCI-H295R cell line related to steroidogenesis.

The current study was intended to evaluate impacts of both iron (Fe) enrichment and overload (in the form of ferrous sulphate heptahydrate, FeSO4.7H2O) on ultrastructural characteristics of human adrenocarcinoma NCI-H295R cell line. Here, the NCI-H295R cells were treated with 0, 3.90, and 1000 µM FeSO4.7H2O, and consequently proceeded for purposes of ultrastructural studies. Micrographs taken under transmission electron microscope (TEM) were investigated from the qualitative and quantitative (unbiased stereological approaches) aspects, and obtained findings were compared among the three groups of the cells. The ultrastructural features related to the steroidogenic process were found to be similar between the untreated and both Fe-exposed cell populations, with conspicuous mitochondria with well-defined lamellar cristae (creating clusters of varying sizes in the regions of increased energy demands) and concentric whorls of smooth endoplasmic reticulum (SER) being the most noticeable characteristics. The precise estimates of the component (volume, surface) fractions of the nucleus, mitochondria, and lipid droplets (LDs), as well as of the nucleus/cytoplasm (N/C) ratio have revealed close similarities (P > 0.05) in all cell groups investigated. Nonetheless, the low concentration of FeSO4.7H2O exhibited beneficial action on ultrastructural organization of the NCI-H295R cells. In effect, these cells were distinguished by mitochondria with smoother surfaces and clearer outlines, higher density of thin, parallel lamellar cristae (deeply extending into the mitochondrial matrix), and more widespread distribution of fine SER tubules as compared to the control ones, all of them suggesting higher level of energy requirements and metabolic activity, and more intensive rate of steroidogenesis. Interestingly, no obvious ultrastructural modifications were observed in the NCI-H295R cells treated with high FeSO4.7H2O concentration. This finding can be linked to either an adaptive ultrastructural machinery of these cells to cope with the adverse effect of the element or to insufficient dose of FeSO4.7H2O (1000 µM) to induce ultrastructural signs of cytotoxicity. Purposefully, the results of the current study complement our previous paper dealing with impacts of FeSO4.7H2O on the NCI-H295R cell viability and steroidogenesis at the molecular level. Hence, they fill a knowledge gap considering structure-function coupling in this cellular model system upon the metal exposure. This integrated approach can enhance our understanding of the cellular responses to Fe enrichment and overload which can be helpful for individuals with reproductive health concerns.

Agronomic Investigation of Spray Dispersion of Metal-Based Nanoparticles on Sunflowers in Real-World Environments.

In environmental and agronomic settings, even minor imbalances can trigger a range of unpredicted responses. Despite the widespread use of metal-based nanoparticles (NPs) and new bio-nanofertilizers, their impact on crop production is absent in the literature. Therefore, our research is focused on the agronomic effect of spray application of gold nanoparticles anchored to SiO2 mesoporous silica (AuSi-NPs), zinc oxide nanoparticles (ZnO-NPs), and iron oxide nanoparticles (Fe3O4-NPs) on sunflowers under real-world environments. Our findings revealed that the biosynthetically prepared AuSi-NPs and ZnO-NPs were highly effective in enhancing sunflower seasonal physiology, e.g., the value of the NDVI index increased from 0.012 to 0.025 after AuSi-NPs application. The distribution of leaf trichomes improved and the grain yield increased from 2.47 t ha-1 to 3.29 t ha-1 after ZnO-NPs application. AuSi-NPs treatment resulted in a higher content of essential linoleic acid (54.37%) when compared to the NPs-free control (51.57%), which had a higher determined oleic acid. No NPs or residual translocated metals were detected in the fully ripe sunflower seeds, except for slightly higher silica content after the AuSi-NPs treatment. Additionally, AuSi-NPs and NPs-free control showed wide insect biodiversity while ZnO-NPs treatment had the lowest value of phosphorus as anti-nutrient. Contradictory but insignificant effect on physiology, yield, and insect biodiversity was observed in Fe3O4-NPs treatment. Therefore, further studies are needed to fully understand the long-term environmental and agricultural sustainability of NPs applications.

Modifications in Ultrastructural Characteristics and Redox Status of Plants under Environmental Stress: A Review.

The rate of global environmental change is unprecedented, with climate change causing an increase in the oscillation and intensification of various abiotic stress factors that have negative impacts on crop production. This issue has become an alarming global concern, especially for countries already facing the threat of food insecurity. Abiotic stressors, such as drought, salinity, extreme temperatures, and metal (nanoparticle) toxicities, are recognized as major constraints in agriculture, and are closely associated with the crop yield penalty and losses in food supply. In order to combat abiotic stress, it is important to understand how plant organs adapt to changing conditions, as this can help produce more stress-resistant or stress-tolerant plants. The investigation of plant tissue ultrastructure and subcellular components can provide valuable insights into plant responses to abiotic stress-related stimuli. In particular, the columella cells (statocytes) of the root cap exhibit a unique architecture that is easily recognizable under a transmission electron microscope, making them a useful experimental model for ultrastructural observations. In combination with the assessment of plant oxidative/antioxidative status, both approaches can shed more light on the cellular and molecular mechanisms involved in plant adaptation to environmental cues. This review summarizes life-threatening factors of the changing environment that lead to stress-related damage to plants, with an emphasis on their subcellular components. Additionally, selected plant responses to such conditions in the context of their ability to adapt and survive in a challenging environment are also described.

Cornelian Cherry (Cornus mas) Powder as a Functional Ingredient for the Formulation of Bread Loaves: Physical Properties, Nutritional Value, Phytochemical Composition, and Sensory Attributes.

In the current study, Cornelian cherry powder (CCP, Cornus mas) was investigated as a functional ingredient for bread production. Experimental bread loaves were prepared using five levels of CCP (0, 1, 2, 5, and 10% w/w) to replace wheat flour in bread formulation. The final products were analyzed regarding their proximate composition, content of selected biologically active substances, antioxidant activity (AA), volume, and sensory attributes. Increasing the incorporation of CCP led to significantly (p < 0.05) higher concentrations of carbohydrate, ash, energetic value, total polyphenols, phenolic acids and AA, and reduced fat and protein contents (p < 0.05). Moreover, up to 5% addition of CCP positively affected the volume (642.63 ± 7.24 mL) and specific volume (2.83 ± 0.02 cm3/g) of bread loaves, which were significantly (p < 0.05) higher compared to the control (no addition of CCP; 576.99 ± 2.97 mL; 2.55 ± 0.002 cm3/g). The sensory attributes chewiness, crumb springiness, bitterness, and sourness had lower scores (p < 0.05) in bread formulated with 10% CCP compared to the control. Overall, results show that the bread loaves produced with up to 5% CCP addition were considered the preferred formulation among the experimental samples tested, taking into consideration their composition, bioactive content, sensory, and physical properties.

Impact of Freeze- and Spray-Drying Microencapsulation Techniques on ß-Glucan Powder Biological Activity: A Comparative Study.

The study compares the impact of freeze- and spray-drying (FD, SD) microencapsulation methods on the content of ß-glucan, total polyphenols (TP), total flavonoids (TF), phenolic acids (PA), and antioxidant activity (AA) in commercially ß-glucan powder (Pleurotus ostreatus) using maltodextrin as a carrier. Morphology (scanning electron microscopy- SEM), yield, moisture content (MC), and water activity (aw) were also evaluated in the samples. Our examinations revealed significant structural differences between powders microencapsulated by the drying methods. As compared to non-encapsulated powder, the SD powder with yield of 44.38 ± 0.55% exhibited more reduced (p < 0.05) values for aw (0.456 ± 0.001) and MC (8.90 ± 0.44%) than the FD one (yield: 27.97 ± 0.33%; aw: 0.506 ± 0.002; MC: 11.30 ± 0.28%). In addition, the highest values for ß-glucan content (72.39 ± 0.38%), TPC (3.40 ± 0.17 mg GAE/g), and TFC (3.07 ± 0.29 mg QE/g) have been detected in the SD powder. Our results allow for the conclusion that the SD microencapsulation method using maltodextrin seems to be more powerful in terms of the ß-glucan powder yield and its contents of ß-glucan, TP, and TF as compared to the FD technique.

Human adrenocortical carcinoma cell line (NCI-H295R): An in vitro screening model for the assessment of endocrine disruptors' actions on steroidogenesis with an emphasis on cell ultrastructural features.

Cell lines as an in vitro model for xenobiotic screening and toxicity studies provide a very important tool in the field of scientific research at the level of molecular pathways and gene expression. Good cell culture practice and intracellular characterization, as well as physiological properties of the cell line are of critical importance for in vitro reproductive toxicity testing of various endocrine-disrupting chemicals. The NCI-H295R, human adrenocarcinoma cell line, is the most widely used in vitro cellular system to study the human adrenal steroidogenic pathway at the level of hormone production and gene expression, as it expresses genes that encode for all the key enzymes for steroidogenesis. In this review, we aim to highlight the information considering the origin, development, physiological and ultrastructural characteristics of the NCI-H295R cell line. The review also creates a broad overview of the cell line usage in various range of studies related to the steroidogenesis issues. To our best knowledge, the paper provides the first report of quantitative data (ex novo) from stereological estimates of component (volume, surface) densities of nuclei, mitochondria, and lipid droplets of the NCI-H295R cells. Such ultrastructural measurements can be valuable in the assessment of underlying mechanisms of changes in the cell steroid hormone production induced by the action of diverse endocrine disruptors. Thus, they can significantly contribute to complexity of structure-function relationships in association with steroidogenesis.

Selected Physico-Chemical, Nutritional, Antioxidant and Sensory Properties of Wheat Bread Supplemented with Apple Pomace Powder as a By-Product from Juice Production.

The present article aimed to study the effects of four selected concentrations (1%, 2%, 5%, and 10%) of apple pomace powder (APP), obtained from juice production, on the nutritional value and selected physico-chemical, antioxidant, and sensory properties of wheat bread. We have found that the ash and total carbohydrate contents, total polyphenols content, and antioxidant activity of the supplemented bread loaves were markedly higher (p < 0.05) as compared to the control ones. On the other hand, values for protein and fat contents and loaf volume in APP-containing bread samples were statistically lower (p < 0.05). Finally, sensory evaluation revealed no significant differences in all tested attributes between the investigated groups of bread samples. The current results suggest that 10% APP addition appears to be an attractive ingredient applied to bread formulation to obtain a bakery product with high nutritional value and required qualitative and sensory properties. In such a manner, apple pomace as by-products from apple juice processing can be efficiently utilized in an eco-friendly way by the food industry to decrease unnecessary waste and environmental pollution.

The Cryopreserved Sperm Traits of Various Ram Breeds: Towards Biodiversity Conservation.

The aim of our research was to compare three Slovak sheep breeds in the quality parameters of cryopreserved sperm. The ejaculates of Slovak Dairy (SD), Native Wallachian (NW), and Improved Wallachian (IW) sheep rams (n = 12) were collected by electro-ejaculation. Heterospermic samples were created from suitable ejaculates, separately for each breed (at least 90% of total and 80% of progressive motility). Samples were equilibrated in a Triladyl® diluent and frozen by automated freezing. Sperm samples were subjected to the motility, morphology, (CASA), viability and apoptosis (DRAQ7/Yo-Pro-1), fertilizing capability (penetration/fertilization test (P/F) in vitro) and acrosomal status (transmission electron microscopy) assays before freezing and after thawing. It was found that there were no significant differences (p < 0.05) between the evaluated breeds in motility, viability, apoptosis, morphological properties, and fertilizing ability of cryopreserved sperm. Significant differences occurred in acrosomal status. Our results demonstrate that the use of the selected cryopreservation protocol is suitable for at least three different sheep breeds, which can greatly benefit the biodiversity protection and simplifies the creation of an animal genetic resources gene bank.

Assessment of Chemical Composition and Anti-Penicillium Activity of Vapours of Essential Oils from Abies Alba and Two Melaleuca Species in Food Model Systems.

The possibilities of the practical utilization of essential oils (EOs) from various plant species in the food industry have attracted the attention of the scientific community. Following our previous studies, the antifungal activities of three further commercial EOs, Melaleuca armillaris subsp. armillaris (rosalina; REO), Melaleuca quinquenervia (niaouli; NEO), and Abies alba (fir; FEO), were evaluated in the present research in respect to their chemical profiles, over four different concentrations, 62.5 µL/L, 125 µL/L, 250 µL/L, and 500 µL/L. The findings revealed that the major compounds of REO, NEO, and FEO were linalool (47.5%), 1,8-cineole (40.8%), and α-pinene (25.2%), respectively. In vitro antifungal determinations showed that the inhibition zones of a Penicillium spp. mycelial growth ranged from no inhibitory effectiveness (00.00 ± 00.00 mm) to 16.00 ± 1.00 mm, indicating a very strong antifungal activity which was detected against P. citrinum after the highest REO concentration exposure. Furthermore, the in situ antifungal efficacy of all EOs investigated was shown to be dose-dependent. In this sense, we have found that the highest concentration (500 µL/L) of REO, NEO, and FEO significantly reduced (p < 0.05) the growth of all Penicillium strains inoculated on the bread, carrot, and potato models. These results indicate that the investigated EOs may be promising innovative agents in order to extend the shelf life of different types of food products, such as bread, carrot and potato.

Rabbit Endothelial Progenitor Cells Derived From Peripheral Blood and Bone Marrow: An Ultrastructural Comparative Study.

The present study was designed to compare the ultrastructure of early endothelial progenitor cells (EPCs) derived from rabbit peripheral blood (PB-EPCs) and bone marrow (BM-EPCs). After the cells had been isolated and cultivated up to passage 3, microphotographs obtained from transmission electron microscope were evaluated from qualitative and quantitative (unbiased stereological approaches) points of view. Our results revealed that both cell populations displayed almost identical ultrastructural characteristics represented by abundant cellular organelles dispersed in the cytoplasm. Moreover, the presence of very occasionally occurring mature endothelial-specific Weibel­Palade bodies (WPBs) confirmed their endothelial lineage origin. The more advanced stage of their differentiation was also demonstrated by the relatively low nucleus/cytoplasm (N/C) ratios (0.41 ± 0.19 in PB-EPCs; 0.37 ± 0.25 in BM-EPCs). Between PB-EPCs and BM-EPCs, no differences in proportions of cells occupied by nucleus (28.13 ± 8.97 versus 25.10 ± 11.48%), mitochondria (3.71 ± 1.33 versus 4.23 ± 1.00%), and lipid droplets (0.65 ± 1.01 versus 0.36 ± 0.40%), as well as in estimations of the organelles surface densities were found. The data provide the first quantitative evaluation of the organelles of interest in PB-EPCs and BM-EPCs, and they can serve as a research framework for understanding cellular function.

Chemical Composition, Antioxidant, In Vitro and In Situ Antimicrobial, Antibiofilm, and Anti-Insect Activity of Cedar atlantica Essential Oil.

The present study was designed to evaluate commercial cedar essential oil (CEO), obtained by hydrodistillation from cedar wood, in relationship to its chemical composition and antioxidant, in vitro and in situ antimicrobial, antibiofilm, and anti-insect activity. For these purposes, gas chromatography-mass spectrometry, DPPH radical-scavenging assay, agar and disc diffusion, and vapor phase methods were used. The results from the volatile profile determination showed that δ-cadinene (36.3%), (Z)-ß-farnesene (13.8%), viridiflorol (7.3%), and himachala-2,4-diene (5.4%) were the major components of the EO chemical constitution. Based on the obtained results, a strong antioxidant effect (81.1%) of the CEO was found. CEO is characterized by diversified antimicrobial activity, and the zones of inhibition ranged from 7.33 to 21.36 mm in gram-positive and gram-negative bacteria, and from 5.44 to 13.67 mm in yeasts and fungi. The lowest values of minimal inhibition concentration (MIC) were noted against gram-positive Micrococcus luteus (7.46 µL/mL) and against yeast Candida krusei (9.46 µL/mL). It seems that the vapor phase of CEO can inhibit the growth of the microscopic filamentous fungi of the genus Penicillium according to in situ antifungal analysis on bread, carrots, and celery. This finding confirms the impact of CEO on the change in the protein structure of older biofilms of Pseudomonas fluorescens and Salmonella enterica subsp. enterica. Insecticidal activity of a vapor phase has also been demonstrated against Pyrrhocoris apterus. CEO showed various advantages on antimicrobial activity, and it is an ideal substitute for food safety.

Antioxidant and antimicrobial activities of fruit extracts from different fresh chili peppers.

BACKGROUND: The aim of the current study was to assess the antimicrobial and antioxidant potential of ethanol extracts obtained from the fruit of five species of fresh chili pepper, Capsicum (C.) baccatum L. (Aji Amarillo), C. chinense (Fidalgo Roxa), C. annuum (Cherry Chocolate), C. pubescens (Rocoto Orange) and C. frutescens (Peruvian Purple). METHODS: To obtain the ethanol extracts, accelerated solvent extraction (ASE) was applied. DPPH assay was used to determine the antioxidant activity of the extract samples. The disc diffusion method was used to measure antimicrobial activity against nine investigated microorganism species. RESULTS: The tested extract samples exhibited DPPH radical scavenging activities ranging from 0.24 ±0.01 (Peruvian Purple) to 0.72 ±0.02 (Aji Amarillo) mg TEAC∙g–1 dw. The differences between all the varieties were statistically significant (P < 0.05; except for the Cherry Chocolate and Rocoto Orange), and the potential of antioxidant capacity increased in the following manner: Peruvian Purple < Fidalgo Roxa < Rocoto Orange < Cherry Chocolate < Aji Amarillo. The results from the antimicrobial evaluation showed that the Capsicum extracts had no uniform inhibition activity against tested gram-negative, gram-positive bacteria, and yeast. Specifically, Aji Amarillo fruit extract revealed the strongest antimicrobial activity against S. pneumoniae (6.33 ±0.58 mm), followed by Cherry Chocolate against S. pneumoniae (5.33 ±0.58 mm), Rocoto Orange against S. enterica (5.27 ±0.58 mm), Fidalgo Roxa against C. albicans (4.67 ±0.58 mm), and Peruvian Purple against S. pneumoniae (4.57 ±0.58 mm). CONCLUSIONS: Considering these results, Capsicum spp. can be used as a source of novel antioxidant and antimicrobial compounds.

Chemical Composition, In Vitro and In Situ Antimicrobial and Antibiofilm Activities of Syzygium aromaticum (Clove) Essential Oil.

The essential oil of Syzygium (S.) aromaticum (CEO) is known for its good biological activity. The aim of the research was to evaluate in vitro and in situ antimicrobial and antibiofilm activity of the essential oil produced in Slovakia. The main components of CEO were eugenol 82.4% and (E)-caryophyllene 14.0%. The antimicrobial activity was either weak or very strong with inhibition zones ranging from 4.67 to 15.78 mm in gram-positive and gram-negative bacteria and from 8.22 to 18.56 mm in yeasts and fungi. Among the tested bacteria and fungi, the lowest values of MIC were determined for Staphylococcus (S.) aureus and Penicillium (P.) expansum, respectively. The vapor phase of CEO inhibited the growth of the microscopic filamentous fungi of the genus Penicillium when tested in situ on bread. The strongest effect of mycelia inhibition in a bread model was observed against P. expansum at concentrations of 250 and 500 µL/mL. The best antimicrobial activity of CEO in the carrot model was found against P. chrysosenum. Differences between the mass spectra of Bacillus (B.) subtilis biofilms on the tested surfaces (wood, glass) and the control sample were noted from the seventh day of culture. There were some changes in mass spectra of Stenotrophomonas (S.) maltophilia, which were observed in both experimental groups from the fifth day of culture. These findings confirmed the impact of CEO on the protein structure of older biofilms. The findings indicate that, besides being safe and sensorially attractive, S. aromaticum has antimicrobial activity, which makes it a potential substitute for chemical food preservatives.

Thymus vulgaris Essential Oil and Its Biological Activity.

Thymus vulgaris essential oil has potential good biological activity. The aim of the research was to evaluate the biological activity of the T. vulgaris essential oil from the Slovak company. The main components of T. vulgaris essential oil were thymol (48.1%), p-cymene (11.7%), 1,8-cineole (6.7), γ-terpinene (6.1%), and carvacrol (5.5%). The antioxidant activity was 85.2 ± 0.2%, which corresponds to 479.34 ± 1.1 TEAC. The antimicrobial activity was moderate or very strong with inhibition zones from 9.89 to 22.44 mm. The lowest values of MIC were determined against B. subtilis, E. faecalis, and S. aureus. In situ antifungal analysis on bread shows that the vapor phase of T. vulgaris essential oil can inhibit the growth of the microscopic filamentous fungi of the genus Penicillium. The antimicrobial activity against S. marcescens showed 46.78-87.80% inhibition at concentrations 62.5-500 µL/mL. The MALDI TOF MS analyses suggest changes in the protein profile of biofilm forming bacteria P. fluorescens and S. enteritidis after the fifth and the ninth day, respectively. Due to the properties of the T. vulgaris essential oil, it can be used in the food industry as a natural supplement to extend the shelf life of the foods.

In vitro effect of ferrous sulphate on bovine spermatozoa motility parameters, viability and Annexin V-labeled membrane changes.

The aim of this study was to assess the dose- and time-dependent in vitro effects of ferrous sulphate (FeSO4.7H2O) on the motility parameters, viability, structural and functional activity of bovine spermatozoa. Spermatozoa motility parameters were determined after exposure to concentrations (3.90, 7.80, 15.60, 31.20, 62.50, 125, 250, 500 and 1000 µM) of FeSO4.7H2O using the SpermVisionTM CASA (Computer Assisted Semen Analyzer) system in different time periods. Cell viability was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) assay, and the Annexin V-Fluos was applied to detect the membrane integrity of spermatozoa. The initial spermatozoa motility showed increased average values at all experimental concentrations compared to the control group (culture medium without FeSO4.7H2O). After 2 h, FeSO4.7H2O stimulated the overall percentage of spermatozoa motility at the concentrations of ≤ 125 µM. However, experimental administration of 250 µM of FeSO4.7H2O significantly (P < 0.001) decreased the spermatozoa motility but had no negative effect on the cell viability (P < 0.05) (Time 2 h). The lowest viability was noted after the addition of ≥ 500 µM of FeSO4.7H2O (P < 0.001). The concentrations of ≤ 62.50 µM of FeSO4.7H2O markedly stimulated (P < 0.001) spermatozoa activity after 24 h of exposure, while at high concentrations of ≥ 500 µM of FeSO4.7H2O the overall percentage of spermatozoa motility was significantly inhibited (P < 0.001) and it elicited cytotoxic action. Fluorescence analysis confirmed that spermatozoa incubated with higher concentrations (≥ 500 µM) of FeSO4.7H2O displayed apoptotic changes, as detected in head membrane (acrosomal part) and mitochondrial portion of spermatozoa. Moreover, the highest concentration and the longest time of exposure (1000 µM of FeSO4.7H2O; Time 6 h) induced even necrotic alterations to spermatozoa. These results suggest that high concentrations of FeSO4.7H2O are able to induce toxic effects on the structure and function of spermatozoa, while low concentrations may have the positive effect on the fertilization potential of spermatozoa.

Thymus serpyllum Essential Oil and Its Biological Activity as a Modern Food Preserver.

The aim of this study was to analyze the chemical composition and biological and antibiofilm activity of the essential oil (EO) of Thymus serpyllum with the use of a MALDI-TOF MS Biotyper. The main compounds of the EO were thymol, 18.8%; carvacrol, 17.4%; o-cymene, 15.4%; and geraniol, 10.7%. It was found that free-radical scavenging activity was high. The highest antimicrobial activity was observed against Pseudomonas aeruginosa, Salmonella enteritidis, and biofilm-forming bacteria. The changes in the biofilm structure after T. serpyllum EO application confirmed the inhibitory action and the most pronounced effect was observed on Bacillus subtilis biofilm. The antifungal activity of the vapor phase was the most effective against Penicillium crustosum. T. serpyllum should be a suitable alternative to synthetic antioxidants as well as antimicrobials. The EO of T. serpyllum can be used in the vapor phase in the storage of root vegetables as well as a growth inhibitor of Penicillium on bread.

In Vitro Antimicrobial Activity of Lavender, Mint, and Rosemary Essential Oils and the Effect of Their Vapours on Growth of Penicillium spp. in a Bread Model System.

The chemical composition, antioxidant activity, and antimicrobial properties of three commercially available essential oils: rosemary (REO), lavender (LEO), and mint (MEO), were determined in the current study. Our data revealed that the major components of REO, MEO, and LEO were 1,8-cineole (40.4%), menthol (40.1%), and linalool acetate (35.0%), respectively. The highest DPPH radical-scavenging activity was identified in MEO (36.85 ± 0.49%) among the investigated EOs. Regarding antimicrobial activities, we found that LEO had the strongest inhibitory efficiencies against the growth of Pseudomonas aeruginosa and Candida (C.) tropicalis, MEO against Salmonella (S.) enterica, and REO against Staphylococcus (S.) aureus. The strongest antifungal activity was displayed by mint EO, which totally inhibited the growth of Penicillium (P.) expansum and P. crustosum in all concentrations; the growth of P. citrinum was completely suppressed only by the lowest MEO concentration. The lowest minimal inhibitory concentrations (MICs) against S. enterica, S. aureus, and C. krusei were assessed for MEO. In situ analysis on the bread model showed that 125 µL/L of REO exhibited the lowest mycelial growth inhibition (MGI) of P. citrinum, and 500 µL/L of MEO caused the highest MGI of P. crustosum. Our results allow us to make conclusion that the analysed EOs have promising potential for use as innovative agents in the storage of bakery products in order to extend their shelf-life.

Mitochondria: A worthwhile object for ultrastructural qualitative characterization and quantification of cells at physiological and pathophysiological states using conventional transmission electron microscopy.

Mitochondria are highly dynamic intracellular organelles with ultrastructural heterogeneity reflecting the behaviour and functions of the cells. The ultrastructural remodelling, performed by the counteracting active processes of mitochondrial fusion and fission, enables the organelles to respond to diverse cellular requirements and cues. It is also an important part of mechanisms underlying adaptation of mitochondria to pathophysiological conditions that challenge the cell homeostasis. However, if the stressor is constantly acting, the adaptive capacity of the cell can be exceeded and defective changes in mitochondrial morphology (indicating the insufficient functionality of mitochondria or development of mitochondrial disorders) may appear. Beside qualitative description of mitochondrial ultrastructure, stereological principles concerning the estimation of alterations in mitochondrial volume density or surface density are invaluable approaches for unbiased quantification of cells under physiological or pathophysiological conditions. In order to improve our understanding of cellular functions and dysfunctions, transmission electron microscopy (TEM) still remains a gold standard for qualitative and quantitative ultrastructural examination of mitochondria from various cell types, as well as from those experienced to different stimuli or toxicity-inducing factors. In the current study, general morphological and functional features of mitochondria, and their ultrastructural heterogeneity related to physiological and pathophysiological states of the cells are reviewed. Moreover, stereological approaches for accurate quantification of mitochondrial ultrastructure from electron micrographs taken from TEM are described in detail.

Copper affects steroidogenesis and viability of human adrenocortical carcinoma (NCI-H295R) cell line in vitro.

Copper is an environmental risk factor, which has various effects on reproductive endocrinology. In this study human adrenocortical carcinoma (NCI-H295R) cell line was used as an in vitro biological model to study the effect of copper sulfate (CuSO4.5H2O) on steroidogenesis and cytotoxicity. The cell cultures were exposed to different concentrations (3.90, 62.50, 250, 500, 1000 µM) of CuSO4.5H2O and compared to control group (medium without CuSO4.5H2O). Cell viability was measured by the metabolic activity assay. Quantification of sexual steroid production directly from the medium was performed by ELISA assay. Following 48 h culture of NCI-H295R cell line in the presence of CuSO4.5H2O a dose-dependent depletion of progesterone release was observed even at the lower concentrations of CuSO4.5H2O. The lowest levels of progesterone were detected in groups with the higher doses (≥ 250 µM) of CuSO4.5H2O, which elicited significant cytotoxic action. Testosterone production decreased significantly, and this decline was more prominent in comparison to that of progesterone. The lowest release of testosterone was recorded at 1000 µM of CuSO4.5H2O. The cytotoxic effect of CuSO4.5H2O was evident at all concentrations used in the study. The presented data suggest that copper has detrimental effects on sexual steroid hormones and consecutively on reproductive physiology.