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.

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.

Effect of Black Cumin Cake Addition on the Chemical Composition, Glycemic Index, Antioxidant Activity, and Cooking Quality of Durum Wheat Pasta.

Pasta is a good carrier for plant enrichment substances due to its popularity among consumers. The purpose of the study was to investigate the functional potential and optimize the recipe of pasta made from durum semolina with the addition of black cumin cake at the level of 5, 10, 15, 20, and 25%. The use of black cumin cake resulted in a statistically significant (p ≤ 0.05) increase in the content of protein, fat, ash, and fiber, including both the insoluble and soluble fractions. A reduction in the digestible carbohydrate content, in vitro starch hydrolysis index (HI), was observed. Pasta with a reduced glycemic index (GI) compared to the semolina control was obtained. The content of polyphenols, including flavonoids, in the cake-enriched pasta increased significantly (p ≤ 0.05), which resulted in higher antioxidant activity against DPPH. The increase in the iron content was over 2.5 times higher in the sample with the 25% addition of black cumin cake than in the control sample. The functional addition significantly (p ≤ 0.05) increased the loss of dry matter and influenced the cooking time of pasta.

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.

Biological Activity and Antibiofilm Molecular Profile of Citrus aurantium Essential Oil and Its Application in a Food Model.

The main aim of the study was to investigate the chemical composition, antioxidant, antimicrobial, and antibiofilm activity of Citrus aurantium essential oil (CAEO). The biofilm profile of Stenotrophonomonas maltophilia and Bacillus subtilis were assessed using the mass spectrometry MALDI-TOF MS Biotyper and the antibiofilm activity of Citrus aurantium (CAEO) was studied on wood and glass surfaces. A semi-quantitative composition using a modified version was applied for the CAEO characterization. The antioxidant activity of CAEO was determined using the DPPH method. The antimicrobial activity was analyzed by disc diffusion for two biofilm producing bacteria, while the vapor phase was used for three penicillia. The antibiofilm activity was observed with the agar microdilution method. The molecular differences of biofilm formation on different days were analyzed, and the genetic similarity was studied with dendrograms constructed from MSP spectra to illustrate the grouping profiles of S. maltophilia and B. subtilis. A differentiated branch was obtained for early growth variants of S. maltophilia for planktonic cells and all experimental groups. The time span can be reported for the grouping pattern of B. subtilis preferentially when comparing to the media matrix, but without clear differences among variants. Furthermore, the minimum inhibitory doses of the CAEO were investigated against microscopic fungi. The results showed that CAEO was most active against Penicillium crustosum, in the vapor phase, on bread and carrot in situ.

Minimal residual disease prior to allogeneic hematopoietic cell transplantation in acute myeloid leukemia: a meta-analysis.

Minimal residual disease prior to allogeneic hematopoietic cell transplantation has been associated with increased risk of relapse and death in patients with acute myeloid leukemia, but detection methodologies and results vary widely. We performed a systematic review and meta-analysis evaluating the prognostic role of minimal residual disease detected by polymerase chain reaction or multiparametric flow cytometry before transplant. We identified 19 articles published between January 2005 and June 2016 and extracted hazard ratios for leukemia-free survival, overall survival, and cumulative incidences of relapse and non-relapse mortality. Pre-transplant minimal residual disease was associated with worse leukemia-free survival (hazard ratio=2.76 [1.90-4.00]), overall survival (hazard ratio=2.36 [1.73-3.22]), and cumulative incidence of relapse (hazard ratio=3.65 [2.53-5.27]), but not non-relapse mortality (hazard ratio=1.12 [0.81-1.55]). These associations held regardless of detection method, conditioning intensity, and patient age. Adverse cytogenetics was not an independent risk factor for death or relapse. There was more heterogeneity among studies using flow cytometry-based than WT1 polymerase chain reaction-based detection (I2=75.1% vs. <0.1% for leukemia-free survival, 67.8% vs. <0.1% for overall survival, and 22.1% vs. <0.1% for cumulative incidence of relapse). These results demonstrate a strong relationship between pre-transplant minimal residual disease and post-transplant relapse and survival. Outcome heterogeneity among studies using flow-based methods may underscore site-specific methodological differences or differences in test performance and interpretation.

Minimal residual disease detectable by quantitative assessment of WT1 gene before allogeneic stem cell transplantation in patients in first remission of acute myeloid leukemia has an impact on their future prognosis.

Overall 42 patients (pts) transplanted in hematological CR1 were retrospectively analyzed. Median follow-up was 15 months (range 2-77). The expression of WT1 gene was measured according to the European Leukaemia Net recommendations. At the time of allogeneic stem cell transplantation (allo-SCT) 29 pts were WT1-negative and 13 pts were WT1-positive. In the univariate analysis, significantly better results were observed in the group of WT1 neg in terms of progression-free survival (in three yr 77% vs. 27%, p = 0.001). In multivariate analysis, the only significant feature in terms of better OS was WT1 negativity (p = 0.029). Our results show that minimal residual disease status measured by quantitative assessment of WT1 gene in acute myeloid leukemia pts in CR1 significantly affects their future prognosis after allo-SCT.

Substantially elevated C-reactive protein (CRP), together with low levels of procalcitonin (PCT), contributes to diagnosis of fungal infection in immunocompromised patients.

PURPOSE: Serum procalcitonin (PCT) has become a routinely utilized parameter with a high prediction value of the severity of bacterial infectious complications and their immediate outcomes. Whereas the utility of PCT in differentiating between bacterial and viral infection is generally accepted, its significance in fungal infections has yet to be determined. The aim of the study was to determine the role of PCT testing in patients at high risk for invasive fungal infections. METHODS: Immunocompromised hematological patients undergoing cyclic chemotherapy treatment or allogeneic hemopoietic stem cell transplantation with infectious complications in which the infectious agents were identified during the disease course were evaluated. In patients with bacterial infection, positive hemocultures were documented, and in patients with fungal infection, the presence of either proven or probable disease was confirmed according to Ascioglu criteria. C-reactive protein (CRP) and PCT were prospectively assessed from the day following fever onset, for four consecutive days. RESULTS: Overall, 34 patients were evaluated, 21 with bacterial and 13 with fungal infections. Significant elevations of CRP concentrations (i.e., above the upper normal limit) were observed in all patients, with a tendency toward higher levels in bacterial (both gram-positive [Gr+] and Gr-negative [Gr-]) than in fungal infections. PCT levels were significantly elevated in patients with bacterial infections (e.g., predominantly in Gr- compared to Gr+), whereas in patients with fungal infections, we identified minimal or no PCT elevations, p < 0.01. For the fungal infections, according to constructed receiver operating characteristic curves, a combination of PCT <0.5 µg/L and CRP 100-300 mg/L offers the best specificity, sensitivity and positive and negative predictive values (81, 85, 73, and 89 %, respectively). CONCLUSION: Altogether, our data suggest that the finding of substantially elevated CRP combined with low PCT in immunocompromised patients may indicate systemic fungal infection. The use of this combination might simplify the diagnostic process, which otherwise can often be lengthy and arduous.

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.

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.

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.

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.

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.

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.

WT1 Gene Expression in Peripheral Blood Before and After Allogeneic Stem Cell Transplantation is a Clinically Relevant Prognostic Marker in AML - A Single-center 14-year Experience.

BACKGROUND: This work summarizes our experience with WT1 monitoring before and after allogeneic hematopoietic stem cell transplantation (allo-HSCT). PATIENTS AND METHODS: The expression of WT1 gene was measured by real-time polymerase chain reaction in peripheral blood according to the European Leukemia Net recommendations. Between May 2005 and August 2019, we analyzed 147 consecutive patients with acute myeloid leukemia with high WT1 expression at diagnosis, transplanted in first (CR1) or second (CR2) complete remission. RESULTS: At the time of allo-HSCT, 107 patients had WT1-normal expression (WT1 ≤ 50 copies), and 40 patients had WT1-high expression. The median follow-up was 21 months. The estimated 5-year overall survival and event-free survival was significantly better in the WT1-normal cohort (65% and 57% vs. 37% and 25%; P = .0003 and P < .0001, respectively) and 5-year cumulative incidence of relapse was significantly lower in the WT1-normal group (19% vs. 53%; P < .0001). Five-year non-relapse mortality was not significantly different (20% and 23%). Multivariate analysis revealed WT1-high expression and acute graft-versus-host disease grade 3/4 as significantly negative prognostic factors for OS. Overall, 49 patients developed WT1 molecular relapse in the post-transplant period; in 14 cases, the therapeutic intervention was done. In all but 1 relapsed patient where WT1 minimal residual disease (MRD) was monitored (38 patients), we detected WT1-high levels (sensitivity of 97%). CONCLUSION: The results of the analysis confirmed our previous experience that WT1 status before allo-HSCT is a strong prognostic factor for both OS and relapse risk. In addition, we confirmed the usefulness of this marker for MRD monitoring after allo-HSCT. The main advantage is the possibility of frequent MRD monitoring in peripheral blood and early bone marrow examination based on WT1-high expression.

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.

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.

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.

WT1 Expression in Peripheral Blood at Diagnosis and During the Course of Early Consolidation Treatment Correlates With Survival in Patients With Intermediate and Poor-Risk Acute Myeloid Leukemia.

BACKGROUND: Up to 55% of non-APL acute myeloid leukemias (AML) lack a molecular target suitable for standardized disease monitoring. We aimed to evaluate the prognostic significance of WT1 gene expression at early stages of intensive treatment. PATIENTS AND METHODS: A total of 106 consecutive patients with intermediate and high-risk AML who had WT1 expression at diagnosis >500 copies/104ABL and who achieved remission after 1 to 2 cycles of induction treatment were analyzed. WT1 expression was measured in peripheral blood using a standardized European LeukemiaNet method. Overexpression was defined as >50 copies/104ABL. The median follow-up was 30 months. RESULTS: Patients with normal versus high WT1 expression after 2 cycles of chemotherapy had overall survival (OS) at 3 years of 66% versus 41% (P = .01); event-free survival (EFS) 45% versus 22% (P = .01). Prognostic significance of WT1 expression after 2 cycles of treatment was maintained in the group of patients treated with chemotherapy alone without hematopoietic stem cell transplantation in first line treatment (OS 70% vs. 36%, P = .02; EFS 35% vs. 0%, P = .03). Significant prognostic factors for EFS on multivariate analysis were the achievement of molecular remission (<50 copies of WT1) at any time during treatment (hazard ratio [HR] 0.47, P = .04) and increased WT1 expression after 2 cycles of chemotherapy (HR 2.0, P = .03). CONCLUSION: Increased WT1 expression after 2 cycles of chemotherapy is a negative prognostic factor for survival. WT1 remains a valuable molecular marker in AML without any leukemia-specific mutation, especially if next generation sequencing and/or digital polymerase chain reaction are not routinely available.

Antioxidant, Antimicrobial and Antibiofilm Activity of Coriander (Coriandrum sativum L.) Essential Oil for Its Application in Foods.

The aim of this study was to assess the chemical composition, antioxidant, antimicrobial and antibiofilm activity of the Coriandrum sativum essential oil. Changes in the biofilm profile of Stenotropomonas maltophilia and Bacillus subtilis were studied using MALDI-TOF MS Biotyper on glass and wooden surfaces. The molecular differences of biofilms in different days were observed as well. The major volatile compounds of the coriander essential oil in the present study were ß-linalool 66.07%. Coriander essential oil radical scavenging activity was 51.05% of inhibition. Coriander essential oil expressed the strongest antibacterial activity against B. subtilis followed by S. maltophilia and Penicillium expansum. The strongest antibiofilm activity of the coriander essential oil was found against S. maltophilia. A clearly differentiated branch was obtained for early growth variants of S. maltophilia in case of planktonic cells and all experimental groups and time span can be reported for the grouping pattern of B. subtilis preferentially when comparing to the media matrix, but without clear differences among variants. The results indicate that coriander was effective against the tested Penicillium expansum in the vapor phase after 14 days with MID50 367.19 and MID90 445.92 µL/L of air.