Studies on the Effects of Fermentation on the Phenolic Profile and Biological Activity of Three Cultivars of Kale.

Fermentation is used not only to preserve food but also to enhance its beneficial effects on human health and achieve functional foods. This study aimed to investigate how different treatments (spontaneous fermentation or fermentation with the use of starter culture) affect phenolic content, antioxidant potential, and cholinesterase inhibitory activity in different kale cultivars: 'Halbhoner Grüner Krauser', 'Scarlet', and 'Nero di Toscana'. Chosen samples were further tested for their protective potential against the Caco-2 cell line. HPLC-MS analysis revealed that the fermentation affected the composition of polyphenolic compounds, leading to an increase in the content of rutin, kaempferol, sinapinic, and protocatechuic acids. In general, kale cultivars demonstrated various antioxidant activities, and fermentation led to an increase in total phenolic content and antioxidant activity. Fermentation boosted anti-cholinesterase activity most profoundly in 'Nero di Toscana'. Extracts of spontaneously fermented 'Scarlet' (SS) and 'Nero di Toscana' (NTS) showed cytoprotective properties, as revealed by the malondialdehyde (MDA), lactate dehydrogenase (LDH), superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH) assays. Additionally, strong anti-inflammatory activity of NTS was shown by decreased release of cytokines IL-1ß and TNF-α. Collectively, the conducted studies suggest fermented kale cultivars as a potential source for functional foods.

Assessment of the genomic stability of calves obtained from artificial insemination and OPU/IVP in vitro fertilization.

Damage to genetic material and errors in the functioning of cellular mechanisms disturb genome stability and integrity. Assessment of genomic stability in animals is a very important aspect of breeding work. Unfortunately, harmful instabilities affect the functioning, health and reproductive processes of animals. Obtaining healthy calves is a priority, whatever methods of reproductive biotechnology are applied. The aim of the study was to assess the genomic stability of calves obtained from artificial insemination and OPU/IVP in vitro fertilization. The genomic stability of the calves was evaluated using the comet, sister chromatid exchange, and fragile sites assays. Damage to the genetic material of calves obtained by two reproductive biotechnologies was identified. Identification of instability in animals can be a valuable tool in breeding work and accelerate breeding progress.

Anti-Inflammatory, Antioxidant, and Neuroprotective Effects of Polyphenols-Polyphenols as an Element of Diet Therapy in Depressive Disorders.

Depressive disorders can affect up to 350 million people worldwide, and in developed countries, the percentage of patients with depressive disorders may be as high as 10%. During depression, activation of pro-inflammatory pathways, mitochondrial dysfunction, increased markers of oxidative stress, and a reduction in the antioxidant effectiveness of the body are observed. It is estimated that approximately 30% of depressed patients do not respond to traditional pharmacological treatments. However, more and more attention is being paid to the influence of active ingredients in food on the course and risk of neurological disorders, including depression. The possibility of using foods containing polyphenols as an element of diet therapy in depression was analyzed in the review. The possibility of whether the consumption of products such as polyphenols could alleviate the course of depression or prevent the progression of it was also considered. Results from preclinical studies demonstrate the potential of phenolic compounds have the potential to reduce depressive behaviors by regulating factors related to oxidative stress, neuroinflammation, and modulation of the intestinal microbiota.

Pleiotropic Potential of Evernia prunastri Extracts and Their Main Compounds Evernic Acid and Atranorin: In Vitro and In Silico Studies.

Evernia prunastri is a lichen widely distributed in the Northern Hemisphere. Its biological properties still need to be discovered. Therefore, our paper focuses on studies of E. prunastri extracts, including its main metabolites evernic acid (EA) or atranorin (ATR). Phytochemical profiles using chromatographic analysis were confirmed. The antioxidant activity was evaluated using in vitro chemical tests and in vitro enzymatic cells-free tests, namely superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GR), and catalase (CAT). The anti-inflammatory potential using cyclooxygenase-2 (COX-2) and hyaluronidase were determined. The neuroprotective potential using acetylcholinesterase, (AChE), butyrylcholinesterase (BChE), and tyrosinase (Tyr) was estimated. The hypoglycemic activity was also confirmed (α-glucosidase). Principal component analysis was performed to determine the relationship between the biological activity of extracts. The inhibitory effect of EA and ATR on COX-2 AChE, BChE, Tyr, and α-glucosidase was evaluated using molecular docking techniques and confirmed for EA and ATR (besides α-glucosidase). The penetration of EA and ATR from extracts through the blood-brain barrier was confirmed using the parallel artificial membrane permeability assay blood-brain barrier test. In conclusion, depending on chemical surroundings and the concentration, the E. prunastri extracts, EA or ATR, showed attractive pleiotropic properties, which should be further investigated.

Basal cell carcinoma and rosacea: coincidence or relationship?

Introduction: The relationship between rosacea and basal cell carcinoma (BCC) is still not known. Almost all reports questioning the relation between BCC and rosacea are focused on rhinophyma. Aim: To examine the possible connection of BCC and rosacea. Material and methods: 110 patients with BCC located on the face treated surgically in the Dermatosurgery Unit in 2020-2021 were included in the study. A trained dermatologist assessed the presence or absence of concomitant rosacea and assessed its severity according to the Investigator's Global Assessment (IGA). Results: Fifty-one (46.4%) subjects with BCC and concomitant clinically diagnosed rosacea were found. In our group 35.5% of women had BCC located on the cheeks, while in men most cancers were located on the nose (55%, n = 11), however this difference has not reached statistical significance. Conclusions: The incidence of rosacea in our population was undoubtedly higher, comparing to the normative population. Both rosacea and BCC share the same risk factors - UV exposure, the potential association should not be neglected.

Translation and validation of the Polish language version of the Teenagers Quality of Life questionnaire (T-QoL).

Introduction: Skin diseases affect patients at any age, but as each period in life is different, tools used to assess quality of life impairment should be adjusted according to the particular age group. Adolescence is a unique time, when young individuals go through many changes, making them especially vulnerable to stress. Aim: Translation and validation of a Polish language version of the Teenagers Quality of Life questionnaire (T-QoL) questionnaire. Material and methods: T-QoL was translated following international guidelines. A group of 34 dermatological patients, aged 12-19 years old, with various skin diseases were given the T-QoL as well as the CDLQI or DLQI to complete. They were also asked to complete the T-QoL questionnaire for the second time after 3-5 days. Statistical analysis of the results was performed. Results: The Polish version of T-QoL is internally consistent (Cronbach α 0.893 for the whole questionnaire). Moreover, it presents very good convergent validity (ICC = 0.864). No statistically significant differences between each question were noticed between the first and second time of completing the form. T-QoL scores correlated significantly with DLQI (p = 0.008, r = 0.636) and CDLQI (p < 0.001, r = 0.777) scores. Conclusions: The Polish version of the T-QoL questionnaire is a reliable instrument with adequate convergent validity, consistency and reproducibility. It can be successfully used to measure quality of life impairment among teenagers.

The Distribution of Glucosinolates in Different Phenotypes of Lepidium peruvianum and Their Role as Acetyl- and Butyrylcholinesterase Inhibitors-In Silico and In Vitro Studies.

The aim of the study was to present the fingerprint of different Lepidium peruvianum tuber extracts showing glucosinolates-containing substances possibly playing an important role in preventinting dementia and other memory disorders. Different phenotypes of Lepidium peruvianum (Brassicaceae) tubers were analysed for their glucosinolate profile using a liquid chromatograph coupled with mass spectrometer (HPLC-ESI-QTOF-MS/MS platform). Qualitative analysis in 50% ethanolic extracts confirmed the presence of ten compounds: aliphatic, indolyl, and aromatic glucosinolates, with glucotropaeolin being the leading one, detected at levels between 0-1.57% depending on phenotype, size, processing, and collection site. The PCA analysis showed important variations in glucosinolate content between the samples and different ratios of the detected compounds. Applied in vitro activity tests confirmed inhibitory properties of extracts and single glucosinolates against acetylcholinesterase (AChE) (15.3-28.9% for the extracts and 55.95-57.60% for individual compounds) and butyrylcholinesterase (BuChE) (71.3-77.2% for the extracts and 36.2-39.9% for individual compounds). The molecular basis for the activity of glucosinolates was explained through molecular docking studies showing that the tested metabolites interacted with tryptophan and histidine residues of the enzymes, most likely blocking their active catalytic side. Based on the obtained results and described mechanism of action, it could be concluded that glucosinolates exhibit inhibitory properties against two cholinesterases present in the synaptic cleft, which indicates that selected phenotypes of L. peruvianum tubers cultivated under well-defined environmental and ecological conditions may present a valuable plant material to be considered for the development of therapeutic products with memory-stimulating properties.

FGFR1-4 RNA-Based Gene Alteration and Expression Analysis in Squamous Non-Small Cell Lung Cancer.

While fibroblast growth factor receptors (FGFRs) are involved in several biological pathways and FGFR inhibitors may be useful in the treatment of squamous non-small cell lung cancer (Sq-NSCLC), FGFR aberrations are not well characterized in Sq-NSCLC. We comprehensively evaluated FGFR expression, fusions, and variants in 40 fresh-frozen primary Sq-NSCLC (stage IA3−IV) samples and tumor-adjacent normal tissues using real-time PCR and next-generation sequencing (NGS). Protein expression of FGFR1−3 and amplification of FGFR1 were also analyzed. FGFR1 and FGFR4 median gene expression was significantly (p < 0.001) decreased in tumors compared with normal tissue. Increased FGFR3 expression enhanced the recurrence risk (hazard ratio 4.72, p = 0.029), while high FGFR4 expression was associated with lymph node metastasis (p = 0.036). Enhanced FGFR1 gene expression was correlated with FGFR1 protein overexpression (r = 0.75, p = 0.0003), but not with FGFR1 amplification. NGS revealed known pathogenic FGFR2,3 variants, an FGFR3::TACC3 fusion, and a novel TACC1::FGFR1 fusion together with FGFR1,2 variants of uncertain significance not previously reported in Sq-NSCLC. These findings expand our knowledge of the Sq-NSCLC molecular background and show that combining different methods increases the rate of FGFR aberrations detection, which may improve patient selection for FGFRi treatment.

Biological Activity of an Epilobium angustifolium L. (Fireweed) Infusion after In Vitro Digestion.

The biological activity of an in vitro digested infusion of Epilobium angustifolium (fireweed) was examined in a model system of intestinal epithelial and colon cancer tissues. The content of selected phenolic compounds in the digested aqueous extract of fireweed was determined using HPLC-ESI-QTOF-MS/MS. Biological activity was examined using the human colon adenocarcinoma cell lines HT-29 and CaCo-2 and the human colon epithelial cell line CCD 841 CoTr. Cytotoxicity was assessed by an MTT assay, a Neutral Red uptake assay, May-Grünwald-Giemsa staining, and a label-free Electric Cell-Substrate Impedance Sensing cytotoxicity assay. The effect of the infusion on the growth of selected intestinal bacteria was also examined. The extract inhibited the growth of intestinal cancer cells HT-29. This effect can be attributed to the activity of quercetin and kaempferol, which were the most abundant phenolic compounds found in the extract after in vitro digestion. The cytotoxicity of the fireweed infusion was dose-dependent. The highest decrease in proliferation (by almost 80%) compared to the control was observed in HT-29 line treated with the extract at a concentration of 250 µg/mL. The fireweed infusion did not affect the growth of beneficial intestinal bacteria, but it did significantly inhibit E. coli. The cytotoxic effect of the fireweed extract indicates that it does not lose its biological activity after in vitro digestion. It can be concluded that the fireweed infusion has the potential to be used as a supporting agent in colon cancer therapy.

Major life changing decision profile: Creation of the Polish language version.

Chronic diseases have long-term consequences and can affect individuals' life course. The aim of this study was to create the Polish language version of a questionnaire estimating the impact of the disease on important life decisions-the major life changing decision profile (MLCDP). The translation of the MLCDP followed international guidelines. The created Polish language version of the questionnaire was administered to 32 nephrology and dermatology ward inpatients. To assess its properties, statistical analysis of the results obtained was conducted. The Polish language version of the MLCDP demonstrated very good internal consistency with a Cronbach α coefficient of 0.84. The questionnaire presented excellent test-retest reliability, established with a coefficient ICC of 0.97. The Polish language version of MLCDP has shown high internal consistency and reproducibility, and can be used effectively to assess the cumulative impact of the disease by indicating the number of major life decisions affected by chronic disease.

Cosmetic Procedure Screening Questionnaire (COPS): creation and validation of the Polish language version.

INTRODUCTION: Body dysmorphic disorder (BDD) is a disabling mental disorder characterized by excessive preoccupation with appearance. Trying to fix imagined defects many individuals with BDD search for aesthetic dermatology treatments. Due to omitting preliminary evaluation for BDD in subjects undergoing cosmetic procedures and lack of proper diagnostic tools among this group of individuals, the results of such interventions may face their disapproval and disappointment. AIM: To translate and validate the Polish version of a Cosmetic Procedure Screening Questionnaire (COPS), which can be used in a cosmetic procedure setting to screen patients suspected to be suffering from BDD. MATERIAL AND METHODS: Both forward and backward translations of the original English version of the questionnaire to Polish were performed in accordance with international standards. The validation was conducted on 33 individuals undergoing aesthetic procedures, who completed the questionnaire twice with 3-6 days' interval. Moreover, the subjects were also asked to fill the Polish versions of BIQLI (Body Image Quality of Life Inventory) and HADS (Hospital Anxiety and Depression Scale) for convergent validity procedure. RESULTS: The Polish version of COPS demonstrated good internal consistency (Cronbach α coefficient value of 0.76) and reproducibility (Intraclass Correlation Coefficient, ICC, of 0.79). COPS correlated strongly with BIQLI (r = -0.66, p < 0.01) as well as with HADS, in both depression and anxiety subscales (r = 0.68, p < 0.01 and r = 0.66, p < 0.01, respectively). CONCLUSIONS: The Polish version of the COPS questionnaire showed sufficient internal consistency and reliability. It can be used for BDD screening among the Polish speaking subjects undergoing aesthetic dermatology procedures.

Family Reported Outcome Measure - 16 (FROM-16): Creation, Reliability and Reproducibility of the Polish Language Version.

Chronic diseases not only have a direct influence on patients' quality of life, but can also affect the life of family members. The aim of this study was to create the Polish language version of a questionnaire estimating impact of disease on quality of family life: the Family Reported Outcome Measure - 16 (FROM-16). A standard forward and backward translation procedure was used to convert the original English version of FROM-16 into the Polish language. Creation of the Polish version was performed in a group of 30 patients' family members. The Polish language version of FROM-16 showed very good internal consistency reliability, the Cronbach α coefficient was 0.89. Reproducibility level was established with an intraclass correlation coefficient of 0.98. The Polish language version of FROM-16 can potentially be used as a tool to assess quality of life of patients' family members.

Honey as the Potential Natural Source of Cholinesterase Inhibitors in Alzheimer's Disease.

Alzheimer's disease (AD) is a progressive neurodegenerative disease characterized, among others by abnormal levels of acetyl- and butyrylcholinesterase in the brain. In this study, 47 types of Polish honeys were examined (using colorimetric method) as a source of acetyl- and butyrylcholinesterase inhibitors. The highest potential for AChE inhibition was observed in the case of buckwheat honey (39.51% inhibition), while multi-floral honey showed the highest capacity for BChE inhibition (39.76%). Our study revealed that honeys can be a rich source of cholinesterase inhibitors and therefore may play a role in AD treatment.

Human Epididymis Protein 4 (HE4) in Patients with Small-Cell Lung Cancer.

BACKGROUND: The aim of the present study was to compare the diagnostic utility of HE4 with NSE, ProGRP, CYFRA 21-1, CEA, and CA 125 and evaluate their prognostic value in patients with small-cell lung cancer (SCLC). METHODS: HE4, ProGRP, NSE, CYFRA 21-1, CEA, and CA 125 assays were performed in 63 patients with smallcell lung cancer (limited disease (LD) - 41, extensive disease (ED) - 22) and in 66 individuals of the reference group. RESULTS: Area under the ROC curves for HE4, ProGRP, NSE, CA 125, CYFRA 21-1, and CEA were 0.884, 0.923, 0.826, 0.796, 0.739, and 0.704, respectively. The tumor marker serum concentrations were associated with tumor stage (HE4, ProGRP, NSE, CYFRA 21-1, CEA), and disease progression occurred within one year (HE4, ProGRP, NSE, CYFRA 21-1). The tumor advancement, performance status, gender and tumor markers, except CEA and CA 125, were significantly associated with survival. Independent, unfavourable prognostic factors included extensive disease (HR 4.14, p < 0.0001) and NSE concentration above 35 g/l (HR 2.62, p = 0.0009). CONCLUSIONS: Diagnostic utility of HE4 was similar to that of NSE and ProGRP. Complementary to NSE, determination of HE4 seems to be helpful in evaluation of SCLC patients' prognosis.

The Effect of Bromodeoxyuridine on Spontaneous Sister Chromatid Exchange Frequency in Rabbit (Oryctolagus cuniculus) and Coypu (Myocastor coypu) Chromosomes.

The sister chromatid exchange test is regarded as a highly sensitive cytogenetic assay. It measures chromosome sensitivity to particular damage factors and provides information on control and repair mechanism performance. It is instrumental in the early identification of the effects of noxious factors present in the habitat. This investigation was aimed at identifying sister chromatid exchange sites in coypu and rabbit chromosomes, as well as determining the spontaneity of the process by applying different BrdU doses. The chromosomes were obtained from an in vitro culture of blood lymphocytes, supplemented with 4 different BrdU doses: 0.25/0.5/1.0/2.5 µg/ml in order to identify spontaneous sister chromatid exchanges in both animal species. The chromosomes were stained according to the FPG method. Spontaneous SCEs were observed in coypu at a concentration of 1.0 µg/ml, and in rabbits at 0.5 µg/ml. The mean SCE/cell incidence was 1.41±1.15 in coypu, and 2.69±2.14 in rabbits. Differences in SCE incidence were identified between the analysed animal species and the applied BrdU doses.

[The comet assay as a method of identifying chromosomes instability]. / Test kometowy jako metoda identyfikacji niestabilnosci chromosomów.

The basic method for analyzing the degree of DNA fragmentation caused by genotoxic factors is gel electrophoresis of single cells (single cell gel electrophoresis), also called the comet assay. The comet assay enables the analysis of the level of several different DNA modifications. The basic testing procedure has been only slightly modified. This method helps identify single-strand and double-strand DNA cracks, as well as any chemical and enzymatic modifications that can potentially turn into cracks in DNA or chromatids. The comet assay makes it possible to detect DNA damage at the level of single cells. It can be employed in analyses of any tissues which provide cellular suspensions. Analysed cells are submerged in agarose on a microscope slide. DNA is what is left after proteins have been broken down. The slide is then subjected to electrophoresis and stained with a fluorescent dye. A "comet-like" image is obtained. The "head" is the cell fixation site prior to lysis; the "tail" represents damaged DNA fragments. The extent of DNA damage is reflected in the length of the tail and the amount of DNA contained in it. The assay finds research applications in the following fields: genetic toxicology, monitoring of DNA repair following chemotherapy and radiotherapy, ecotoxicology, animal and human nourishment, biomonitoring of genotoxicity, epidemiology and assessment of material deposited in sperm and blood banks.

Age-dependent change in the morphology of nucleoli and methylation of genes of the Nucleolar Organizer Region in Japanese quail Coturnix japonica) model (Temminck and Schlegel, 1849) (Galliformes: Aves).

Nucleoli are the product of the activity of nucleolar organizer regions (NOR) in certain chromosomes. Their main functions are the formation of ribosomal subunits from ribosomal protein molecules and the transcription of genes encoding rRNA. The aim of the study was to determine the shape of nucleoli and analyse methylation in the gene RN28S in the spermatocytes of male Japanese quail (Coturnixjaponica) in two age groups. Nucleoli were analysed in cells of the first meiotic prophase. Their number and shape were determined and they were classified as regular, irregular or defragmented. In the cells of the young birds no defragmented nucleoli were observed, with regular and irregular nucleoli accounting for 97% and 3%, respectively. In the cells of older birds no regular nucleoli were observed, while irregular and defragmented nucleoli accounted for 37% and 67%, respectively. MSP (methylation-specific PCR) showed that the gene RN28S is methylated in both 15-week-old and 52-week-old quails. In recent years an association has been established between nucleolus morphology and cellular ageing processes.

Vegan and Vegetarian Soups Are Excellent Sources of Cholinesterase Inhibitors.

BACKGROUND: The cholinesterase theory stands as the most popular worldwide therapy for Alzheimer's disease (AD). Given the absence of a cure for AD, a plant-based diet has been repeatedly shown as positive in the prevention of AD, including exploring ready-made products in stores and the development of new functional foods. GOAL: This study compared the anti-acetyl- and butyrylcholinesterase activity of thirty-two Polish market soups and five newly formulated soups intended to be functional. Additionally, the research aimed to assess the significance of animal content, distinguishing between vegan and vegetarian options, in cholinesterase inhibition. MATERIALS AND METHODS: The anticholinesterase activity was investigated using a spectrophotometric method, and the inhibitory activity was expressed as % inhibition of the enzyme. The study categorized soups into three groups based on ingredients: those containing animal-derived components, vegetarian soups and vegan soups. RESULTS: Soups exhibited varying levels of activity against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), indicating differences in their compositions. Composition appeared to be the primary factor influencing anticholinesterase activity, as soups within each group showed significant variability in activity levels. While some commercial soups demonstrated notable anticholinesterase activity, they did not surpass the effectiveness of the optimized soups developed in the laboratory. Certain ingredients were associated with higher anticholinesterase activity, such as coconut, potato, onion, garlic, parsley and various spices and herbs. CONCLUSIONS: Vegetarian and vegan soups exhibited comparable or even superior anticholinesterase activity compared to animal-derived soups, highlighting the importance of plant-based ingredients. The study underscores the need for further research to explore the mechanisms underlying the anticholinesterase activity of soups, including the impact of ingredient combinations and processing methods.