Oils from Transgenic Flax Lines as Potential Chemopreventive Agents in Colorectal Cancer.

Colorectal cancer is a major global health concern, and the need for effective chemopreventive agents is paramount. This study aimed to evaluate the potential of oils from transgenically modified flax for the prevention of colorectal cancer, in relation to the oil concertation. Flaxseed oils were obtained from traditional (Nike) and genetically modified flax lines (M and B). Cell viability assays were performed on various cancer cell lines, including colon adenocarcinoma cells. Flaxseed oil B exhibited the strongest anti-proliferative properties compared to the reference drugs and other oils. Additionally, M and B oils showed enhanced accumulation of Rhodamine 123 and increased apoptosis in colorectal cancer cells. M oil exhibited the highest levels of p53 protein. Notably, the tested transgenic oils did not induce metastasis and displayed stronger inhibition of COX-1 compared to COX-2. These data indicate the utility of flaxseed oils, especially from the M line, as adjuvants in colorectal cancer treatment, targeting the colon specifically.

Comparative Analysis of Fused Deposition Modeling and Digital Light Processing Techniques for Dimensional Accuracy in Clear Aligner Manufacturing.

BACKGROUND This study aimed to compare fused deposition modeling (FDM) and digital light processing (DLP) techniques in terms of dimensional accuracy for printing dental models used for the manufacture of clear dental aligners. MATERIAL AND METHODS Based on the intraoral scan of an adult patient, a sequence of 10 aligner models was created using BlueSkyPlan4. The test models (n=30) were fabricated with 2 desktop 3D printers: (DLP) and (FDM) printers. Two groups of samples were created (digitized using a desktop optical scanner). To calculate trueness (n=20) and precision (n=10), printed models were compared to the source files (REF). REF, DLP, and FDM files were superimposed and converted to point clouds. The cloud-to-cloud distances were calculated using CloudCompare software. Using the same algorithm, distortions of models were measured. Data were analyzed using one-way ANOVA and Tukey's post hoc test. RESULTS Significant differences were found between the trueness and precision of DLP and FDM groups. The average calculated trueness of DLP and FDM was 0.096 mm (0.021) (P<0.001) and 0.063 mm (0.024) (P<0.001), respectively. The average calculated precision of DLP and FDM was 0.027 mm (0.003) (P<0.001) and 0.036 mm (0.003) (P<0.001), respectively. A widening (0.158 mmfor DLP and 0.093 mmfor FDM, P=0.05) and twisting (0.03 mmfor DLP and 0.043 mmfor FDM, P=0.05) of the printed models was observed. CONCLUSIONS Both printers had sufficient precision for aligner models manufacturing. FDM showed a higher trueness and this device can be applied as an alternative to DLP. Polymerization shrinkage is a significant factor in decreasing the trueness of DLP printers.

Evaluation of Recovery Methods for Fragaria vesca L. Oil: Characteristics, Stability and Bioactive Potential.

Wild strawberry (Fragaria vesca L.) seed oil (WSO) recovered by two methods-cold pressing (CP) and extraction with supercritical carbon dioxide (SCO2E)-taking into account the different extraction times, was characterized for its composition and quality. The cytotoxicity assessment of WSOs was also carried out using the normal human dermal fibroblast (NHDF) cell line. Tocopherol and total polyphenol contents were significantly higher in WSO recovered by SCO2E, up to 1901.0 and 58.5 mg/kg, respectively, in comparison with CP oil. In CP oil, the highest content of carotenoids and squalene was determined (123.8 and 31.4 mg/kg, respectively). Phytosterol summed up to 5396 mg/kg in WSO collected in 30 min of SCO2E. Moreover, the highest oxidative stability was found for this oil. All studied WSOs were non-cytotoxic in lactate dehydrogenase (LDH) leaching and sulforhodamine B (SRB) assays; however, oils collected by SCO2E in 15 and 30 min were found to be cytotoxic in the tetrazolium salt (MTT) test, with the CC50 at a concentration of 3.4 and 5.5%, respectively. In conclusion, the composition of WSO indicates that, depending on the method of its recovery, seeds can have different bio-potencies and various applications.

Preservation fluids of heritage anatomical specimens - a challenge for modern science. Studies of the origin, composition and microbiological contamination of old museum collections.

Anatomical museums preserve specimens of great historical value and undiscovered scientific potential. However, frequently these collections lack documentation of the techniques of preparation and the composition of preservative substances (conservation principles). This poses a huge problem for the care and preservation of these materials, more so because understanding this issue requires knowledge of fundamentals from different scientific disciplines. The aim of the research was to obtain information about the composition of substances used to preserve historic specimens, as well as to conduct a microbiological assessment of the specimens to detect possible factors causing their deterioration. Furthermore, we wanted to fill an existing gap in the literature, as there is a lack of reports on analytical methods that could be successfully applied by anatomists involved in the daily care of museum collections in human anatomy departments. The starting point was the analysis of the sources and history of the collections, on which basis the choice of research methods was made. Methods based on simple chemical reactions and specialised methods (such as gas chromatography-tandem mass spectrometry, Fourier transform infrared spectroscopy, inductively coupled plasma optical emission spectroscopy) were used in the analyses of the composition of fluids. Microbiological analyses were based on culture and isolation methods, analysis of microscopy slides and matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry analysis. As a result of these analyses, some components of the preservative mixtures and their concentrations were determined. The presence of methanol, ethanol, formaldehyde and glycerol was detected, among other chemicals. The concentrations of these substances were different between the samples and their determination required the use of a variety of methods suitable for the individual components of the preservative mixture. In microbiological tests, both bacteria and fungi were isolated from swabs taken from anatomical specimens. The bacterial flora was less numerous than the fungal flora. Among the bacteria, environmental Gram-positive Bacillus cereus, Bacillus thuringiensis and a rare bacterium of the Cupriavidus genus were isolated, whereas among the fungal organisms, the yeast-like fungi Candida boidinii and Geotrichum silvicola as well as mould fungi Penicillium sp. and Fusarium sp. were detected. However, the microscopic evaluation showed a greater diversity of microorganisms, which may be related to the fact that many environmental bacteria cannot be cultured using classical methods, but can be observed under the microscope. The results of the research made it possible to draw conclusions about the mutual influence of physical, chemical, and microbiological factors on the condition of historical anatomical specimens. In the course of the research, information was obtained on the processes which could have taken place during the storage of these collections. Maintaining the integrity of a container housing a preserved anatomical specimen has a major impact on maintaining the concentration of preservative fluid and keeping the specimen environment sterile. Many conservation procedures for historical specimens carried out nowadays pose a risk of destroying valuable specimens, as well as a health risk for the person carrying out the work. The exploration of the topic of conservation of anatomical specimens, especially those that lack documentation of their origin, is a key issue in current research on historical collections of anatomical specimens.

Overexpression of Bacterial Beta-Ketothiolase Improves Flax (Linum usitatissimum L.) Retting and Changes the Fibre Properties.

Beta-ketothiolases are involved in the beta-oxidation of fatty acids and the metabolism of hormones, benzenoids, and hydroxybutyrate. The expression of bacterial beta-ketothiolase in flax (Linum usitatissimum L.) results in an increase in endogenous beta-ketothiolase mRNA levels and beta-hydroxybutyrate content. In the present work, the effect of overexpression of beta-ketothiolase on retting and stem and fibre composition of flax plants is presented. The content of the components was evaluated by high-performance liquid chromatography, gas chromatography-mass spectrometry, Fourier-transform infrared spectroscopy, and biochemical methods. Changes in the stem cell walls, especially in the lower lignin and pectin content, resulted in more efficient retting. The overexpression of beta-ketothiolase reduced the fatty acid and carotenoid contents in flax and affected the distribution of phenolic compounds between free and cell wall-bound components. The obtained fibres were characterized by a slightly lower content of phenolic compounds and changes in the composition of the cell wall. Based on the IR analysis, we concluded that the production of hydroxybutyrate reduced the cellulose crystallinity and led to the formation of shorter but more flexible cellulose chains, while not changing the content of the cell wall components. We speculate that the changes in chemical composition of the stems and fibres are the result of the regulatory properties of hydroxybutyrate. This provides us with a novel way to influence metabolic composition in agriculturally important crops.

Abscisic Acid-Defensive Player in Flax Response to Fusarium culmorum Infection.

Fusarium culmorum is a ubiquitous soil pathogen with a wide host range. In flax (Linum ussitatissimum), it causes foot and root rot and accumulation of mycotoxins in flax products. Fungal infections lead to huge losses in the flax industry. Moreover, due to mycotoxin accumulation, flax products constitute a potential threat to the consumers. We discovered that the defense against this pathogen in flax is based on early oxidative burst among others. In flax plants infected with F. culmorum, the most affected genes are connected with ROS production and processing, callose synthesis and ABA production. We hypothesize that ABA triggers defense mechanism in flax and is a significant player in a successful response to infection.

Bioactive Compounds of Raspberry Oil Emulsions Induced Oxidative Stress via Stimulating the Accumulation of Reactive Oxygen Species and NO in Cancer Cells.

There are growing interests in the complex combinations of natural compounds that may advance the therapy of cancer. Such combinations already exist in foods, and a good representative is seed oils. Two raspberry oils: cold pressed (ROCOP) and one extracted by supercritical CO2 (ROSCO2) were evaluated for their chemical characteristics and oil emulsions for cell suppression potential against colon adenocarcinoma (LoVo), doxorubicin-resistant colon adenocarcinoma (LoVo/DX), breast cancer (MCF7), doxorubicin-resistant breast cancer (MCF7/DX), and lung cancer (A549) cell lines. The cytotoxicity was also assessed on normal human dermal fibroblasts (NHDFs). With increasing concentration of raspberry oil emulsions (0.5-10%), increasing inhibition of cancer cell viability and proliferation in all of the lines was observed, with different degrees of potency between cancer types and oil tested. ROSCO2 strongly induced free radical production and DNA strand damage in LoVo and MCF7 cells especially doxorubicin-resistant lines. This suggests that ROSCO2 engages and effectively targets the vulnerabilities of the cancer cell. Generally, both ROSCO2 and ROCOP could be a nontoxic support in therapy of selected human cancers.

Chemistry, oxidative stability and bioactivity of oil extracted from Rosa rugosa (Thunb.) seeds by supercritical carbon dioxide.

Rosa rugosa Thunb. seed oil (RR) extracted by supercritical CO2 was investigated. RR chemical composition, radical scavenging effect and oxidative stability were evaluated. RR aqueous emulsions were examined for cell cytotoxicity, proliferation, redox state and migration using mouse embryonic fibroblast Balb/3T3, human dermal fibroblast NHDF cell lines, and on neoplastic cell lines: acute monocytic leukemia THP-1 and lung adenocarcinoma A549. RR total contents of phytosterols, tocopherols, carotenoids and phenolics were 10115.23, 784.16, 40.32 and 10.30 mg/kg, respectively. Rich antioxidant composition of RR was reflected in its high antioxidant activity (2.1 mM/kg Trolox equivalent) as well as oxidative stability (activation energy 105.6 kJ/mol). The RR emulsions led to marked augmentation of the total cell protein content in BALB/3T3 and NHDF cultures, inhibited cancer cell migration and reduced ROS formation. The studied RR oil proved to have a remarkable combination of bioactive compounds and to exert an antioxidative and chemopreventive effects.

Characteristics and Antioxidant Potential of Cold-Pressed Oils-Possible Strategies to Improve Oil Stability.

The relations of the antiradical capacity to oxidative stability parameters and the contents of fatty acids, sterols, tocopherols, phenols, flavonoids, chlorophyll, Cu, and Fe were assessed in 33 cold-pressed seed oils: Walnut (7 brands of oils), rosehip (3), camelina (6), milk thistle (5), flax (6), and pumpkin (6). The antiradical capacity of oils depended strongly on tocopherol contents with a synergistic effect with polyphenols. The efficacy of tocopherols in cold-pressed oils was accompanied by a negative correlation of their antioxidant capacity with the peroxide value increase after 3 months of shelf life. This study also showed a positive correlation between the content of phytosterols and the antiradical capacity in the lipophilic fraction of cold-pressed oils rich in n-3 polyunsaturated fatty acids (PUFAs). Multiple regression analysis identified groups of antioxidants naturally occurring in cold-pressed oils in relation to their fatty acid composition, which added to the cold-pressed oils could provide possible strategies to improve their stability. Achieving high stability is primarily a result of high phytosterol content exceeding the molar ratio of 1:100 for total phytosterols to α-linolenic acid. However, the molar ratios of tocopherols to linoleic acid below 1:2000 and polyphenols to linoleic acid below 1:3000 does not prevent oxidation in oils with the predominance of linoleic acid.

Natural phenolics greatly increase flax (Linum usitatissimum) oil stability.

BACKGROUND: Flaxseed oil is characterized by high content of essential polyunsaturated fatty acids (PUFA) promoted as a human dietary supplement protecting against atherosclerosis. The disadvantage of the high PUFA content in flax oil is high susceptibility to oxidation, which can result in carcinogenic compound formation. Linola flax cultivar is characterized by high linoleic acid content in comparison to traditional flax cultivars rich in linolenic acid. The changes in fatty acid proportions increase oxidative stability of Linola oil and broaden its use as an edible oil for cooking. However one of investigated transgenic lines has high ALA content making it suitable as omega-3 source. Protection of PUFA oxidation is a critical factor in oil quality. The aim of this study was to investigate the impact of phenylpropanoid contents on the oil properties important during the whole technological process from seed storage to grinding and oil pressing, which may influence health benefits as well as shelf-life, and to establish guidelines for the selection of new cultivars. METHODS: The composition of oils was determined by chromatographic (GS-FID and LC-PDA-MS) methods. Antioxidant properties of secondary metabolites were analyzed by DPPH method. The stability of oils was investigated: a) during regular storage by measuring acid value peroxide value p-anisidine value malondialdehyde, conjugated dienes and trienes; b) by using accelerated rancidity tests by TBARS reaction; c) by thermoanalytical - differential scanning calorimetry (DSC). RESULTS: In one approach, in order to increase oil stability, exogenous substances added are mainly lipid soluble antioxidants from the isoprenoid pathway, such as tocopherol and carotene. The other approach is based on transgenic plant generation that accumulates water soluble compounds. Increased accumulation of phenolic compounds in flax seeds was achieved by three different strategies that modify genes coding for enzymes from the phenylpropanoid pathway. The three types of transgenic flax had different phenylpropanoid profiles detected in oil, highly increasing its stability. CONCLUSIONS: We found that hydrophilic phenylpropanoids more than lipophilic isoprenoid compounds determine oil stability however they can work synergistically. Among phenolics the caffeic acid was most effective in increasing oil stability.

Characteristics of rose hip (Rosa canina L.) cold-pressed oil and its oxidative stability studied by the differential scanning calorimetry method.

Two new commercially available high linolenic oils, pressed at low temperature from rose hip seeds, were characterised for their composition, quality and DPPH radical scavenging activity. The oxidative stability of oils was assessed using differential scanning calorimetry (DSC). Phytosterols, tocopherols and carotenoids contents were up to 6485.4; 1124.7; and 107.7 mg/kg, respectively. Phenolic compounds determined for the first time in rose hip oil totalled up to 783.55 µg/kg, with a predominant presence of p-coumaric acid methyl ester. Antiradical activity of the oils reached up to 3.00 mM/kg TEAC. The acid, peroxide and p-anisidine values as well as iron and copper contents indicated good quality of the oils. Relatively high protection against oxidative stress in the oils seemed to be a result of their high antioxidant capacity and the level of unsaturation of fatty acids.

The Antioxidant Activity and Oxidative Stability of Cold-Pressed Oils.

In our study, we characterized the antioxidant activity and oxidative stability of cold-pressed macadamia, avocado, sesame, safflower, pumpkin, rose hip, Linola, flaxseed, walnut, hempseed, poppy, and milk thistle oils. The radical scavenging activity of the non-fractionated fresh oil, as well as the lipophilic and hydrophilic fractions of the oil was determined using a 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay. The fatty acid composition of the fresh and stored oils was analyzed by gas chromatography. The acid value, peroxide value, p-anisidine value and conjugated diene and triene contents in the fresh oils, as well as in those stored throughout the whole period of their shelf life, were measured by CEN ISO methods. The antioxidant activity of the oils expressed as Trolox equivalent antioxidant capacity (TEAC), ranged from 0.17 to 2.32 mM. The lipophilic fractions of the oils were characterized by much higher antioxidant activity than the hydrophilic ones. There were no significant changes in fatty acid composition and only slight changes in the oxidative stability parameters of the oils during their shelf life. Through the assessment of the relationship between antiradical activity and the oxidative stability of oils, it is proposed that a DPPH assay predicts the formation of oxidation products in cold-pressed oils-however, the correlations differ in fractionated and nonfractionated oils.

Mass spectrometric behavior of phenolic acids standards and their analysis in the plant samples with LC/ESI/MS system.

Liquid chromatography coupled to mass spectrometry (MS) with electrospray ionization (ESI) is one of analytical techniques to obtain accurate results of low molecular weight aromatic compounds in biological samples of different origin. The interpretations of mass spectra of these aromatic compounds in the negative spectra registered in the full scan MS mode may be uneasy due to presence of deprotonated molecules [M-H](-) from different co-eluting entities, fragment ions created after the break-up of precursor ions and also ions representing modified molecules clusters. Thus, the first aim of this study was to evaluate general parameters during analysis performed in the full scan MS or MS/MS mode. Secondly, to set general fragmentation rules for aromatic compounds and entities in a complex biological matrix. We established that different groups of low molecular weight phenolic acids form unique adduct ions and additionally registration LC/MS/MS spectra with two different collision energies may allow for differentiating isomeric or isobaric molecules. These findings together with some general fragmentation rules can facilitate identifications of aromatic acids as we outlined in the sample of cold-pressed rose-hip oil and lupine leaves extract.

Maternal dietary fat determines metabolic profile and the magnitude of endocannabinoid inhibition of the stress response in neonatal rat offspring.

Endocannabinoids (eCBs) are products of phospholipid (PL)-derived arachidonic acid (AA) that regulate hypothalamus-pituitary-adrenal axis activity. We hypothesized that differences in the quality and quantity of maternal dietary fat would modulate the PL AA content in the neonatal brain affecting stress responsiveness via differences in eCB production and activity in stress-activated brain areas. Pregnant rats were fed a 5% [control (C)] or 30% fat [high fat (HF)] diet rich in either n-6 (HF-n-6) or n-3 (HF-n-3) fat during the last week of gestation and lactation. Postnatal d 10 offspring were tested for metabolic hormones, AA (n-6) and eCB brain content, and hormonal effects of eCB receptor antagonism (AM251, 1 or 3 mg/kg ip) on stress responses. Like maternal diet, milk from HF-n-3 mothers had a reduced n-6/n-3 fat ratio compared with that of C and HF-n-6 mothers. Hypothalamic and hippocampal levels of PL AA were diet specific, reflecting the maternal milk and dietary n-6/n-3 ratio, with HF-n-3 offspring displaying reduced AA content relative to C and HF-n-6 offspring. Plasma corticosterone and insulin were elevated in HF-fed pups, whereas leptin was increased only in HF-n-6 pups. Basal eCB concentrations were also diet and brain region specific. In C pups, eCB receptor antagonist pretreatment increased stress-induced ACTH secretion, but not in the HF groups. Stress-induced corticosterone secretion was not sensitive to AM251 treatment in HF-n-3 pups. Thus, the nature of preweaning dietary fat differentially influences neonatal metabolic hormones, brain PL AA levels, and eCB, with functional consequences on hypothalamus-pituitary-adrenal axis modulation in developing rat pups.