In this study, boroxine derivative (K₂B₃O₃F₄OH) was tested as an inhibitor of horseradish peroxidase (HRP) by spectrophotometric and electrochemical methods. The activity of horseradish peroxidase ...was first studied under steady-state kinetic conditions by a spectrophotometric method which required the use of guaiacol as a second substrate to measure guaiacol peroxidation. The results of this method have shown that, by changing the concentration of guaiacol as the literature suggests, a different type of inhibition is observed than when changing the concentration of hydrogen peroxide as the substrate. This suggests that guaiacol interferes with the reaction in some way. The electrochemical method involves direct electron transfer of HRP immobilized in Nafion nanocomposite films on a glassy carbon (GC) electrode, creating a sensor with an electro-catalytic response to the reduction of hydrogen peroxide. The electrochemical method simplifies kinetic assays by removing the requirement of reducing substrates.
The cyclic AMP pathway promotes melanocyte differentiation by activating CREB and the cAMP-regulated transcription co-activators 1–3 (CRTC1–3). Differentiation is dysregulated in melanomas, although ...the contributions of CRTC proteins is unclear. We report a selective differentiation impairment in CRTC3 KO melanocytes and melanoma cells, due to downregulation of oculo-cutaneous albinism II (OCA2) and block of melanosome maturation. CRTC3 stimulates OCA2 expression by binding to CREB on a conserved enhancer, a regulatory site for pigmentation and melanoma risk. CRTC3 is uniquely activated by ERK1/2-mediated phosphorylation at Ser391 and by low levels of cAMP. Phosphorylation at Ser391 is constitutively elevated in human melanoma cells with hyperactivated ERK1/2 signaling; knockout of CRTC3 in this setting impairs anchorage-independent growth, migration, and invasiveness, whereas CRTC3 overexpression supports cell survival in response to the mitogen-activated protein kinase (MAPK) inhibitor vemurafenib. As melanomas expressing gain-of-function mutations in CRTC3 are associated with reduced survival, our results suggest that CRTC3 inhibition may provide therapeutic benefit in this setting.
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•CRTC3 knockout mice have decreased fur pigmentation•Melanosome maturation is defective in CRTC3 knockout melanocytes•CREB and CRTC3 promote OCA2 expression in melanocytes•Increased CRTC3 expression/activity in melanomas correlates with reduced survival
MAPK/ERK and cAMP pathways influence differentiation of melanocytes and melanoma progression. Ostojić et al. show that the CREB coactivator CRTC3 is activated by both pathways, leading to increased expression of genes regulating melanin production and cell migration. Decreases in CRTC3 activity result in hypopigmentation and impairment of melanoma oncogenesis.
In this study, high hydrostatic pressure extraction (HHPE) and ultrasound-assisted extraction (UAE) were used for the improvement of pectin and polyphenol recovery from tomato peel waste.
HHPE ...enhanced pectin recovery by 15% after 45 min of recycling, in comparison with the conventional extraction (CE) for 180 min. Similar mass fractions of anhydrouronic acid, total sugars and total phenols were obtained by using both methods. FTIR and 1H NMR data confirmed that chemical composition of pectin obtained by HHPE and CE is identical, therefore it was concluded that the faster HHPE method can be used for its further isolation.
Although depectinized residues subjected to UAE in 70% ethanol for 15 min contained two times lower values of total phenols (1625.7 mg/100 g) than pectinized samples (3643.9 mg/100 g), their quantities are not negligible, considering the fact that they are generated after HHPE. At the end of UAE, the residues were exploited as a source of fatty acids, among which lauric, palmitic and stearic acids are dominant.
In conclusion, by shortening the extraction time using HHPE and UAE, it is possible to efficiently produce two valuable functional ingredients, pectin and polyphenols, and at the same time to reduce peel waste from tomato canning industry, which presents an environmental problem.
Utilizing HHPE and UAE as novel and emerging technologies, and combining them with traditional ones (Soxhlet) is given the solution for sequential isolation of pectin, polyphenols and fatty acids from tomato peel waste, generated by canning factory.
Shortening of extraction time using HHPE and UAE, it is possible to replace the conventional techniques, and achieve efficient production of pectin and polyphenols.
Overall, the extraction methodology proposed in this work could provide two valuable benefits, i.e. the producers could find mode for decreasing of disposal costs of waste and consumer would take opportunity that isolated compounds could be reintroduced into food.
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•Tomato peel waste is utilized for sequential extraction of pectin and polyphenols.•Shortening of time using HHPE and UAE is achieved their efficient production.•HHPE enhanced pectin recovery by 15% after 45 min of recycling.•UAE of depectinized samples for 15 min yielded high quantities of phenols.•At the end of sonication, the residues are utilized as a source of fatty acids.
Triclosan (TCS) is a polychlorinated phenoxy phenol (PCPPs) used as a disinfectant and a broad-spectrum antibacterial and antifungal agent in personal hygiene products. TCS easily forms diphenyl ...ethers and dioxins, which are persistent organic pollutants.
This work used a double approach for the TSC sensing: a) screen-printed (SPE) electrochemical platform for on-site application, modified with lanthanum iron oxide and graphitic carbon nitride composite (LaFeO3/Fe2O3@g-C3N4/SPE); and b) carbon paste electrode (CPE), modified with the same material and used in laboratory conditions. Linear range from 0.1 μM to 10 μM, the limit of detection (LOD) of 29 nM and the limit of quantification (LOQ) of 91 nM were obtained for CP electrode in BRBS pH 8. SPE showed the best analytical parameters in BRBS at pH 3, with a linear range from 0.3 μM to 7 μM, LOD of 0.09 μM and LOQ of 0.28 μM. Furthermore, the influence of potential interferents was investigated and proven to be negligible. Determination of TSC was performed to estimate the environmental impact of this compound as well as the practical usefulness of the proposed sensor in the real sample analysis, confirmed with a HPLC analysis.
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•Creation of a new assembly for TCS detection in personal hygiene products•A sensitive and selective voltammetric approach for single drop TCS detection•Synthesis and characterization of LaFeO3/Fe2O3@g-C3N4 composite materials
Liver phospholipid fatty acid composition depends on the dietary lipid intake and the efficiency of hepatic enzymatic activity. Our study aimed to simultaneously investigate the liver phospholipid ...fatty acid composition in response to chronic linseed, palm, or sunflower oil diets. We used adult female C57/BL6 mice and randomly divided them into control and three groups treated with 25 % dietary oils. Prior to treatment, we analyzed the fatty acid profiles in dietary oils and hepatocytes and, after 100 days, the fatty acid composition in the liver using gas-liquid chromatography. Linseed oil treatment elevated alpha-linolenic, eicosapentaenoic, and docosapentaenoic acids and reduced arachidonic and docosatetraenoic acids, consequently lowering the n-6/n-3 ratio. Palm oil treatment increased linoleic acid and decreased docosahexaenoic acid, contributing to an elevated n-6/n-3 ratio. Sunflower oil treatment elevated total monounsaturated fatty acids by increasing palmitoleic, oleic, and vaccenic acids. The estimated activity of Δ9 desaturase was significantly elevated in the sunflower oil group, while Δ5 desaturase was the highest, and Δ6 desaturase was the lowest after the linseed oil diet. Our findings demonstrate that chronic consumption of linseed, palm, or sunflower oil alters the distribution of liver phospholipid fatty acids differently. Sunflower oil diet elevated total monounsaturated fatty acids, proposing potential benefits for liver tissue health. Considering these outcomes, a substantial recommendation emerges to elevate linseed oil intake, recognized as the principal ALA source, thereby aiding in reducing the n-6/n-3 ratio. Moreover, modifying dietary habits to incorporate specific vegetable oils in daily consumption could substantially enhance overall health.
•Liver phospholipid composition reflects dietary lipid intake and enzymatic activity.•Desirable n-6/n-3 ratio emphasizes the importance of dietary lipid balance.•Vegetable oils are crucial sources of energy and essential fatty acids in diets.•Chronic linseed, palm, or sunflower oil intake alters liver phospholipid composition.•Dietary vegetable oil selection is critical for regulating liver lipid composition.
Occasionally, Hashimoto's thyroiditis (HT) and papillary thyroid carcinoma (PTC) share similar nuclear features. The current study aims to quantify the differences between the investigated specimens ...of HT-associated PTC versus the HT alone, to reduce the subjective experience of an observer, by the use of fractal parameters as well as gray-level co-occurrence matrix (GLCM) textural parameters. We have analyzed 250 segmented nuclei per group (nn = 25 per patient and np = 10 patients per group) using the ImageJ software (NIH, Bethesda, MD, USA) as well as an in-house written code for the GLCM analysis. The mean values of parameters were calculated for each patient. The results demonstrated that the malignant cells from the HT-associated PTC specimens showed lower chromatin fractal dimension (p = 0.0321) and higher lacunarity (p = 0.0038) compared with the corresponding cells from the HT specimens. Also, there was a statistically significant difference between the investigated specimens, in the contrast, correlation, angular second moment, and homogeneity, of the GLCM corresponding to the visual texture of follicular cell chromatin. The differences in chromatin fractal and GLCM parameters could be integrated with other diagnostic methods for the improved evaluation of distinctive features of the HT-associated PTC versus the HT in cytology and surgical pathology specimens.
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•Development of adsorption process for eriochrome black T removal by biosorbent.•Hazelnut shell, a low-cost and green biosorbent showed high effectiveness for EBT.•Physisorption, ...chemisorption and diffusion are studied during the removal process.•Addition of seven interfering ions enhanced the removal efficiency of EBT to 96%.•Hazelnut shell has shown great effectiveness in the removal of six heavy metal ions.
Water pollution with dyes that are non-biodegradable has become a very concerning environmental problem in the world today. Therefore, the aim of this research was to optimize an adsorption process for the removal of one of the most harmful dyes Eriochrome Black T (EBT). For the first time, native hazelnut shells were used as biosorbents in this regard. The biomaterial was analysed by infrared spectroscopy with Fourier transformation, scanning electron microscopy, electron dispersive spectroscopy, Boehm titrations, physicochemical analysis and by determination of zero charge point and cation exchange capacity. Dye concentration was established by UV–Vis spectrophotometer and heavy metals concentration by flame atomic absorption spectrometry (FAAS). Characterization techniques of EBT showed successful removal from wastewater due to the obvious changes in morphological and composition structure of the hazelnut shell. The highest capacity of adsorption (5.8 mg/g) and removal efficiency (75.3%) was found at pH 3, sorbent mass 500 mg, 80 mg/L EBT concentration, 90 min contact time and solution temperature 20 °C. The pseudo-second-order model described the best investigated process and maximum Langmuir’s adsorption capacity was 27.55 mg/g. The interfering effect of seven heavy metals increased removal efficiency to 98% showing a positive impact. Furthermore, the process was found feasible with negative values of Gibbs free energy and exothermic with negative values of enthalpy change (-35.26 kJ/mol). The utilized green and environmentally friendly sorbent under optimized conditions could be used in real textile industries for EBT and heavy metals removal.
Neutrophil extracellular traps (NETs), which are extracellular DNA structures released from neutrophils, are described and characterized for the first time in fish using fluorescent confocal ...microscopy. Confocal images of fish neutrophil suspensions stained with 6′-diamino-2-phenylindole, dihydrochloride DNA fluorescent stain (DAPI) revealed the presence of NETs which appeared as fibrous structures connecting several cells. Co-localization of NETs with neutrophil granular proteins and actin was investigated using specific antibodies and probes. Double staining of neutrophils with SYTOX green and DAPI revealed that SYTOX stain applied to living cells stained extracellular DNA, but not nuclei. NETs are actively released from stimulated living cells, associated with granular proteins, but not with cytoskeleton, and are not a product of nuclear degradation seen in late apoptotic stages. Additionally, a fluorometric microtiter plate assay to quantify the release of NETs was adopted for use with fish neutrophils, and the effect of stress on NETs release was studied. This assay detected the inhibition of DNA release during stress conditions. In summary, NETs were released from living fish kidney neutrophils upon stimulation, characterized using fluorescence DNA-binding dyes, specific antibodies and probes, and quantified using a microtiter plate fluorometric assay that can rapidly measure a large number of samples. Detection of NETs can be used as an additional assay to an existing battery of functional tests, and as a new research model to study the effects of stress, immunomodulators, and diseases.
Larger proportions of chronic obstructive pulmonary disease (COPD) patients are currently overweight or with obesity than underweight, and the combination of COPD and obesity is increasing. The ...purpose of this study was to investigate differences in the body composition, pulmonary function tests, exercise capacity, and health-related quality of life among normal weight, overweight, and obese patients with COPD.
A total of 514 patients with COPD were included in the study. According to the World Health Organization criteria for body mass index, the patients were classified as normal weight, overweight, and obese. Evaluations included fat-free mass, fat-free mass index, phase angle, pulmonary function tests, and 6-minute walk test. Dyspnea was assessed using the modified Medical Research Council dyspnea scale, and the health-related quality of life was evaluated using COPD Assessment Test and St. George's Respiratory Questionnaire. Values were compared among the three groups.
There were 315 male and 199 female patients, with a mean age of 66.7 ± 8.4 years. Fat-free mass, fat-free mass index, and phase angle values were significantly higher in COPD patients with obesity than in other patients (P < .001, P < .001, P < .001). Forced expiratory volume in 1 s, forced expiratory volume in 1 s/forced vital capacity, and diffusing capacity of lung for carbon monoxide value in pulmonary function tests were significantly higher in COPD patients with obesity than in other patients (P = .046, P < .001, P < .001), while the forced vital capacity values were similar in all groups. Exercise capacity (6-min walk test distance), dyspnea symptoms (modified Medical Research Council scale), and health-related quality of life (COPD Assessment Test and St. George's Respiratory Questionnaire) did not differ significantly between groups.
According to our study, obesity has no negative effect on pulmonary function tests, dyspnea perception, exercise capacity, and health-related quality of life.
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) caused a global pandemic and one group of patients has developed a severe form of COVID-19 pneumonia with an urgent need for ...hospitalization and intensive care unit (ICU) admission. The aim of our study was to evaluate the prognostic role of MDW, CRP, procalcitonin (PCT), and lactate in critically ill COVID-19 patients. The primary outcome of interest is the 28 day mortality of ICU patients with confirmed SARS-CoV-2 infection and sepsis (according to Sepsis 3 criteria with acute change in SOFA score ≥ 2 points). Patients were divided into two groups according to survival on the 28th day after admission to the ICU. Every group was divided into two subgroups (women and men). Nonparametric tests (Mann-Whitney) for variables age, PCT, lactate, and MDW were lower than alpha
< 0.05, so there was a significant difference between survived and deceased patients. The Chi-square test confirmed statistically significant higher values of MDW and lactate in the non-survivor group. We found a significant association between MDW, lactate, procalcitonin, and fatal outcome, higher values were reported in the deceased group.
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