Inflammatory and intestinal permeability biomarkers in healthy participants on long term vegan, vegetarian, omnivore and low-carbohydrate high-fat diet.

Vegan, vegetarian and low-carbohydrate high fat (LCHF) diets can all offer several health benefits, if food choices are appropriate. In most studies examining their effects on systemic inflammation, participants were either overweight, on a weight loss programme or not matched for BMI, or had a pre-existing condition such as type 2 diabetes mellitus or hypertension. Little is known about the effects of dietary patterns on healthy and normal weight individuals. The aim of the present study was therefore to assess and directly compare inflammatory and intestinal permeability status in healthy participants following aforementioned or omnivore diet for at least 6 months. In this cross-sectional study, we measured the inflammatory biomarkers IL-6, TNF-α and CRP, and the markers of intestinal permeability LBP and zonulin, along with the analysis of lifestyle aspects, dietary intakes and physical activity, in 89 healthy participants. The groups were matched for sex, age and BMI. There were no differences in any of the measured parameters between the four groups and we found no strong correlations with dietary intakes. Using cluster analysis, participants were divided into eight clusters with more or less favourable inflammatory profiles; all clusters contained representatives of all patterns and all patterns were represented in each cluster. Significant differences between clusters were in the intake of mono-unsaturated fatty acids, ω-3/ω-6 ratio, phase angle and working two shifts. In healthy, normal-weight individuals, inflammatory status therefore does not depend on the dietary pattern itself, but is rather more complexly regulated and associated with dietary and non-dietary factors.

Could Gut Microbiota Composition Be a Useful Indicator of a Long-Term Dietary Pattern?

Despite the known effects of diet on gut microbiota composition, not many studies have evaluated the relationship between distinct dietary patterns and gut microbiota. The aim of our study was to determine whether gut microbiota composition could be a useful indicator of a long-term dietary pattern. We collected data from 89 subjects adhering to omnivorous, vegetarian, vegan, and low-carbohydrate, high-fat diet that were equally distributed between groups and homogenous by age, gender, and BMI. Gut microbiota composition was analyzed with a metabarcoding approach using V4 hypervariable region of the 16S rRNA gene. K-means clustering of gut microbiota at the genus level was performed and the nearest neighbor classifier was applied to predict microbiota clustering classes. Our results suggest that gut microbiota composition at the genus level is not a useful indicator of a subject's dietary pattern, with the exception of a vegan diet that is represented by a high abundance of Prevotella 9. Based on our model, a combination of 26 variables (anthropometric measurements, serum biomarkers, lifestyle factors, gastrointestinal symptoms, psychological factors, specific nutrients intake) is more important to predict an individual's microbiota composition cluster, with 91% accuracy, than the dietary intake alone. Our findings could serve to develop strategies to educate individuals about changes of some modifiable lifestyle factors, aiming to classify them into clusters with favorable health markers, independent of their dietary pattern.

Effects of Four Different Dietary Fibre Supplements on Weight Loss and Lipid and Glucose Serum Profiles during Energy Restriction in Patients with Traits of Metabolic Syndrome: A Comparative, Randomized, Placebo-Controlled Study.

Obesity and its associated complications require various lifestyle changes and treatment options. Dietary supplements are considered an attractive alternative to traditional therapy, mainly because they are accessible to the general population. The aim of this study was to investigate the additive effects of a combination of energy restriction (ER) and four dietary supplements on changes in the anthropometric and biochemical parameters in 100 overweight or obese participants who were randomly assigned one of the dietary fibre supplements containing different dietary fibres or a placebo for 8 weeks. The results confirmed that fibre supplements plus ER significantly (p < 0.01) reduced the body weight, body mass index (BMI), fat mass, and visceral fat and ameliorated the lipid profile and inflammation at 4 and 8 weeks after the start of the study, while in the placebo group, significant differences in some parameters were observed only after 8 weeks of ER. A fibre supplement containing glucomannan, inulin, psyllium, and apple fibre was the most effective at reducing the BMI, body weight, and CRP (p = 0.018 for BMI and body weight and p = 0.034 for CRP compared to placebo at the end of the intervention). Overall, the results suggest that dietary fibre supplements in combination with ER may have additional effects on weight loss and the metabolic profile. Therefore, taking dietary fibre supplements may be a feasible approach to improve weight and metabolic health in obese and overweight individuals.

Habitual low carbohydrate high fat diet compared with omnivorous, vegan, and vegetarian diets.

Background: Dietary patterns which exclude whole food groups, such as vegetarian, vegan and low carbohydrate high fat diet (LCHF), are increasingly popular in general public. When carefully planned, all these diets have some known benefits for health, but concerns are also raised in particular for LCHF. The quality of LCHF diet which individuals follow in real life without supervision is not known. Methods: One hundred thirty healthy individuals with stable body mass following LCHF, vegan, vegetarian and omnivorous diet for at least six months, were compared in a cross-sectional study. Diet was analyzed through 3-day food records and FFQ, anthropometric measurements were performed and serum metabolic biomarkers determined from fasting blood. Results: Participants on LCHF diet had the intakes of micronutrients comparable to other groups, while the intakes of macronutrients differed in line with the definition of each diet. The intakes of saturated fats, cholesterol and animal proteins were significantly higher and the intakes of sugars and dietary fibers were lower compared to other groups. Healthy eating index 2015 in this group was the lowest. There were no differences in the levels of glucose, triacylglycerols and CRP among groups. Total and LDL cholesterol levels were significantly higher in LCHF group, in particular in participants with higher ketogenic ratio. Fatty acids intakes and intakes of cholesterol, dietary fibers and animal proteins explained 40% of variance in total cholesterol level, with saturated fatty acids being the strongest positive predictor and monounsaturated fatty acids a negative predictor. Conclusion: None of the self-advised diets provided all the necessary nutrients in optimal levels. Due to the detected increased levels of serum cholesterols, selection of healthy fat sources, higher intake of dietary fibers and partial replacing of animal sources with plant sources of foods should be recommended to the individuals selecting LCFH dietary pattern.Clinical Trial Registration: ClinicalTrials.gov, identifier NCT04347213.

Impact of Sample Preparation Methods on Single-Cell X-ray Microscopy and Light Elemental Analysis Evaluated by Combined Low Energy X-ray Fluorescence, STXM and AFM.

BACKGROUND: Although X-ray fluorescence microscopy is becoming a widely used technique for single-cell analysis, sample preparation for this microscopy remains one of the main challenges in obtaining optimal conditions for the measurements in the X-ray regime. The information available to researchers on sample treatment is inadequate and unclear, sometimes leading to wasted time and jeopardizing the experiment's success. Many cell fixation methods have been described, but none of them have been systematically tested and declared the most suitable for synchrotron X-ray microscopy. METHODS: The HEC-1-A endometrial cells, human spermatozoa, and human embryonic kidney (HEK-293) cells were fixed with organic solvents and cross-linking methods: 70% ethanol, 3.7%, and 2% paraformaldehyde; in addition, HEK-293 cells were subjected to methanol/ C3H6O treatment and cryofixation. Fixation methods were compared by coupling low-energy X-ray fluorescence with scanning transmission X-ray microscopy and atomic force microscopy. RESULTS: Organic solvents lead to greater dehydration of cells, which has the most significant effect on the distribution and depletion of diffusion elements. Paraformaldehyde provides robust and reproducible data. Finally, the cryofixed cells provide the best morphology and element content results. CONCLUSION: Although cryofixation seems to be the most appropriate method as it allows for keeping cells closer to physiological conditions, it has some technical limitations. Paraformaldehyde, when used at the average concentration of 3.7%, is also an excellent alternative for X-ray microscopy.

Link between emotional and external eating behaviors, peripheral neuropeptide Y, and ß-hydroxybutyrate in participants with obesity on 12-week ketogenic diet.

Objective: Understanding the impact of stress on emotional and external eating behaviors and the psychological and the associated metabolic factors can help in designing subsequent interventions to protect health. In particular, psychological trait-like construct related to eating has been shown to be an important target for intervention. Methods and measures: This study aimed to investigate the biochemical variables associated with a decrease in emotional and external eating behaviors due to 12-week ketogenic diet (12KD) in 35 adult participants (12 males) with obesity. Results: Absolute changes in emotional and external eating were independent of changes in body mass, nutritional intake, and Δ cortisol, but were predicted with increases in serum ß-hydroxybutyrate (BHB) and decreases in serum peripheral neuropeptide Y (pNPY) (all p's < 0.050). Decrease in pNPY was also associated with an increase in BHB but was independent of anthropometrical changes, Δ fasting glucose, and Δ insulin. Conclusion: The reductions in emotional and external eating behaviors in participants with obesity were uniquely predicted by an increase in BHB and a decrease in pNPY after 12KD. In ketosis, emotional and external eating dropped independently of body mass change. Change in pNPY predicted changes in emotional and external eating. The role of BHB in modulating eating behavior should be further explored.

Helichrysum italicum (Roth) G. Don and Helichrysum arenarium (L.) Moench infusions in reversing the traits of metabolic syndrome: a double-blind randomized comparative trial.

Health impairments characteristic for metabolic syndrome such as increased body mass, a dysregulated lipid or glucose profile and elevated blood pressure can be reversed by appropriate lifestyle modifications. Supplementing the normal diet with herbal infusions is a promising strategy. We conducted a randomised double-blind comparative study in which participants with at least two traits of metabolic syndrome consumed an infusion of either Helichrysum italicum subsp. italicum (HI, n = 14) or Helichrysum arenarium (HA, n = 13) daily for 28 days. Anthropometric and biochemical parameters were measured at baseline, at the end of the intervention and after a 2-week washout period. HI infusion consumption had a beneficial effect on anthropometric traits; significant reductions in body weight, body mass index, and visceral and total body fat were observed. In the HA group, there was a greater reduction in serum glucose levels and an improvement in the lipid profile. In both groups, high LDL levels were measured at baseline, but two weeks after the intervention, in 84% of participants in the HA group and 71% in the HI group, the levels were within the reference range. Both interventions caused a decrease in HDL but also improved serum antioxidant properties. Consuming either infusion could thus be recommended as a simple, profitable habit for individuals with traits of metabolic syndrome.

The effect of COVID-19 lockdown on mental health, gut microbiota composition and serum cortisol levels.

The aim of this study was to assess changes in mental health, gut microbiota composition, and stress marker serum cortisol due to COVID-19 lockdown in asymptomatic individuals. Healthy adults participated in anthropometric measurements, blood and stool sample collection pre-lockdown and post-lockdown (n = 38, 63.2% females), lifestyle and psychological questionnaires were included in pre-lockdown measurement and lockdown survey (n = 46, 67.4% females). Subjects reported significantly higher body dissatisfaction (p = 0.007) and anxiety (p = 0.002), and significantly lower positive affect (p = 0.001) during lockdown compared with pre-lockdown. According to perceived stress, 51.6% of females and 20% of males experienced moderate to high stress. This was reflected in serum cortisol levels that significantly increased only in females (p = 0.006) post-lockdown and correlated with perceived stress (p = 0.037) and anxiety (p = 0.031). In addition to psychological measures, changes in gut microbiota composition were observed. Gut microbial alpha diversity significantly decreased (p = 0.033), whereas relative abundance of Proteobacteria significantly increased (p = 0.043) post-lockdown. Depression during lockdown was moderately positively correlated with changes in Bacteroidetes abundance (p = 0.015) and negatively with changes in Firmicutes abundance (p = 0.008). Alistipes abundance post-lockdown was moderately positively correlated with anxiety (p = 0.004) and negative affect (p = 0.005) during lockdown. Despite a small sample size and not being able to perform objective measurements during lockdown, the results confirm the effect of lockdown on mental health and gut microbiota composition that could have a great impact on our health (ClinicalTrials identifier: NCT04347213).

Whole transcriptome expression array analysis of human colon fibroblasts culture treated with Helichrysum italicum supports its use in traditional medicine.

ETHNOPHARMACOLOGICAL RELEVANCE: Helichrysum italicum (HI) is a Mediterranean plant with well-reported use in traditional medicine for a wide range of applications, including digestive and liver disorders, intestinal parasitic infections, wound healing, stomach ache and asthma. However, little is known about the global mechanism behind its pleiotropic activity. AIM OF THE STUDY: The aim of this study was to explain the mechanism behind the previously demonstrated effects of HI and to justify its use in traditional medicine. MATERIALS AND METHODS: A microarray-based transcriptome analysis was used to discover the global transcriptional alterations in primary colon fibroblasts after exposure to HI infusion for 6 h and 24 h. In addition, quantitative real-time PCR was used to verify the microarray results. RESULTS: Altogether we identified 217 differentially expressed genes compared to non-treated cells, and only 8 were common to both treatments. Gene ontology analysis revealed that 24 h treatment with HI infusion altered the expression of genes involved in cytoskeletal rearrangement and cell growth, whereas pathway analysis further showed the importance of interleukin signaling and transcriptional regulation by TP53. For the 6 h treatment only the process of hemostasis appeared in the results of both enrichment analyses. In functional assays, HI infusion increased cell migration and decreased blood clotting and prothrombin time. CONCLUSIONS: With the careful evaluation of the role of individual genes, especially SERPING1, ARHGAP1, IL33 and CDKN1A, represented in the enriched pathways and processes, we propose the main mode of HI action, which is wound healing. In addition to its indirect prevention of diseases resulting from the impaired barrier integrity, HI also effects inflammation and metabolic processes directly, as it regulates genes such as LRPPRC, LIPA, ABCA12, PRKAR1A and ANXA6.

Helichrysum italicum (Roth) G. Don and Helichrysum arenarium (L.) Moench Infusion Consumption Affects the Inflammatory Status and the Composition of Human Gut Microbiota in Patients with Traits of Metabolic Syndrome: A Randomized Comparative Study.

Helichrysum italicum (Roth) G. Don (HI) and Helichrysum arenarium (L.) Moench (HA) are rich in polyphenols and their infusions have beneficial effects for patients with metabolic syndrome. To investigate whether these effects are mediated by the gut microbiota, we analysed the effects of daily consumption of HI or HA infusion on the composition of gut microbiota, inflammatory status, and zonulin, a marker of gut barrier permeability. The study was a randomized, double-blind comparative trial. Thirty participants were randomly assigned to two groups and received either HA or HI tea filter bags, each containing 1 g of dried plant material, for daily consumption lasting 4 weeks. The results show that consumption of both infusions resulted in a reduction of some genera belonging to Firmicutes and in a slight but significant reduction in Shannon diversity index. Consumption of HI infusion significantly reduced serum levels of proinflammatory markers and zonulin alongside with the observed trend of Proteobacteria reduction. It can therefore be concluded that the HI and HA infusions could act as prebiotics and thus improve the intestinal environment. In addition, HI infusion has a positive impact on microbial dysbiosis and gut barrier dysfunction that occur in obesity and metabolic syndrome.

High-Protein Bar as a Meal Replacement in Elite Sports Nutrition: A Pilot Study.

This study was focused on the creation of high-protein bars formulated using whey protein isolate (24%) and soy protein isolate (6%) as the sources of proteins; oat flakes and inulin, both abundant in dietary fibres, and creatine monohydrate and other minor ingredients (vitamin and mineral mixture, potassium sorbate) to achieve the requirements for a meal replacement formula for physically active people. The nutritional profile of the high-protein bar was examined (energy 1215 kJ/288 kcal; protein 34.1 ± 0.20 g, fat 6.01 ± 0.13 g of which was saturated 3.12 ± 0.08 g, fibre 3.10 ± 0.17 g carbohydrate 23.0 ± 0.16 g of which sugars 1.50 ± 0.19 g and starch 21.5 ± 0.11 g in 100 g), and sensory properties with instrumental parameters (texture and colour) were determined and compared with bars commercially available on the market. The created high-protein bar was sensorily acceptable in comparison to other commercially available bars. The dietary intervention study was conducted on elite athletes (professional handball players) to evaluate effects of created versus control bar consumption on their metabolic parameters. The baseline characteristics (mean age, body mass index (BMI), fat mass, muscle mass, lean mass and fat percentage) of the athletes (8) were determined at the start of the study. The cross-over intervention study was organized in two successive phases (5 days each) with a seven-day long washout period between phases. Bars were consumed after the afternoon training unit. Blood samples were collected at the start and the end of the intervention study to analyse the metabolic profiles of the athletes. Serum levels of high-density cholesterol (HDL), low-density cholesterol (LDL) and total cholesterol (HOL), glucose, triacylglycerides (TAG), total and direct bilirubin, creatine kinase (CK), aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) were measured. The results showed that bar consumption significantly decreased serum aspartate transaminase (AST) and lactate dehydrogenase (LDH) and increased total and direct bilirubin levels, suggesting lower exercise-induced muscle damage and increased antioxidative response, respectively. Therefore, it can be concluded that the consumption of the created high-protein bar was able to improve physiological adaptation after training.

Helichrysum italicum ssp. italicum Infusion Promotes Fat Oxidation in Hepatocytes and Stimulates Energy Expenditure and Fat Oxidation after Acute Ingestion in Humans: A Pilot Study.

Helichrysum italicum is an aromatic plant with promising pharmacological activities. Bioactive compounds found in plants represent an important alternative treatment for weight loss and an infusion of H. italicum contains compounds which could have such effect. Our aim was, therefore, to investigate its acute effects on resting energy expenditure (REE) and possible differences in substrate oxidation in a pilot study. A dried, ground plant material of H. italicum ssp. italicum was infused with hot water and chemically characterized with HPLC-MS analysis. Sensory evaluation of herbal tea was performed. A randomized, crossover, controlled pilot study was then conducted on eleven healthy male subjects. The REE and substrate oxidation were measured by indirect calorimetry at baseline and 30 and 120 min after ingestion of infusion or hot water. The expression of genes involved in lipid metabolism was examined in H. italicum infusion-treated hepatocytes. Several phenolic classes were identified in the infusion, caffeoylquinic acids were the most abundant, followed by pyrones and flavonols. A single ingestion of H. italicum infusion significantly increased REE by 4% and fat oxidation by 12% compared to hot water ingestion. A significant 2-fold up-regulation of ß-oxidation-related genes in HepG2 cells, exposed to H. italicum infusion, was detected. This pilot study suggests that H. italicum infusion possesses bioactive substances with potential application in obesity prevention, which could, with additional studies, become an economically interesting novel application of the plant. Clinical trial registration number: NCT04818905.

A Review and Evaluation of the Data Supporting Internal Use of Helichrysum italicum.

Helichrysum italicum is a Mediterranean plant with various pharmacological activities. Despite extensive reports on the bioactivity of the plant, its clinically studied applications have not yet been reviewed. The aim of our study was to gather information on the internal use of H. italicum and its bioactive constituents to determine its efficacy and safety for human use. We reviewed research articles that have not been previously presented in this context and analyzed relevant clinical studies with H. italicum. Cochranelibrary.com revealed six eligible clinical trials with H. italicum that examined indications for pain management, cough, and mental exhaustion. Although the efficacy of H. italicum has been demonstrated both in in vitro tests and in humans, it is difficult to attribute results from clinical trials to H. italicum alone, as it has usually not been tested as the sole component. On the other hand, clinical trials provide positive information on the safety profile since no adverse effects have been reported. We conclude that H. italicum is safe to use internally, while new clinical studies with H. italicum as a single component are needed to prove its efficacy. Based on the recent trend in H. italicum research, further studies are to be expected.

A Comparative Study of the Antioxidative Effects of Helichrysum italicum and Helichrysum arenarium Infusions.

Helichrysum arenarium (L.) Moench (abbrev. as HA) has a long tradition in European ethnomedicine and its inflorescences are approved as a herbal medicinal product. In the Mediterranean part of Europe, Helichrysum italicum (Roth) G. Don (abbrev. as HI) is more common. Since infusions from both plants are traditionally used, we aimed to compare their antioxidative potential using in vitro assays. Two morphologically distinct HI plants, HIa and HIb, were compared to a commercially available HA product. Genetic analysis using microsatellites confirmed a clear differentiation between HI and HA and suggested that HIb was a hybrid resulting from spontaneous hybridization from unknown HI subspecies. High-performance liquid chromatography-mass spectrometry analysis showed the highest amounts of hydroxycinnamic acids and total arzanol derivatives in HIa, whereas HIb was richest in monohydroxybenzoic acids, caffeic acids, and coumarins, and HA contained the highest amounts of flavonoids, especially flavanones. HIa exhibited the highest radical scavenging activity; it was more efficient in protecting different cell lines from induced oxidative stress and in inducing oxidative stress-related genes superoxide dismutase 1, catalase, and glutathione reductase 1. The antioxidative potential of HI was not only dependent on the morphological type of the plant but also on the harvest date, revealing important information for obtaining the best possible product. Considering the superior properties of HI compared to HA, the evaluation of HI as a medicinal plant could be recommended.

The Impact of COVID-19-Related Lockdown on Diet and Serum Markers in Healthy Adults.

Due to limited data about the impact of lockdown on health status, the present study aimed to investigate the impact of COVID-19-related lockdown on changes in dietary habits, physical activity and serum markers in healthy adults. A total of 38 asymptomatic adults aged from 23 to 59 with a normal BMI (22.5 kg/m2) participated in baseline and post-lockdown measurements that included dietary and physical activity assessment, anthropometric measurements and blood samples; and the lockdown survey which included dietary assessment and questionnaires about changes in lifestyle and physical activity. A decreased diet quality during lockdown was observed (Healthy Eating Index reduced from 64.59 to 61.08), which returned to near baseline post-lockdown. Energy intake decreased during lockdown (p = 0.002) and returned to baseline post-lockdown. Despite lower physical activity levels during lockdown (p = 0.035), we observed no significant changes in body composition. However, we observed a significant increase in serum glucose (p = 0.005), total cholesterol (p = 0.003), and low-density lipoprotein (LDL) (p = 0.049) post-lockdown. Increase in serum glucose levels was pronounced in subjects with higher increase in energy intake (p = 0.039), increased omega-6 fatty acids intake (p = 0.016), those who were exposed to several risky contacts (p = 0.018, compared to those with less risky contacts) and those who were not active in nature (p = 0.008, compared to those active in nature). Increased serum LDL was correlated to decreased monounsaturated fatty acids intake (p = 0.028). Within the limits of this preliminary report, changes in serum markers observed among healthy subjects point to a possible impact of COVID-19-related lockdown on adults' health to be confirmed in larger groups.

Assessment of micronutrients in a 12-wk ketogenic diet in obese adults.

OBJECTIVE: A 12-wk ketogenic diet was found to have many beneficial effects in healthy obese adults, but it is not clear if the supply of micronutrients is adequate. METHODS: In 35 adult individuals with body mass index >30, the intakes of minerals and their serum levels were analyzed at baseline and at weeks 4 and 12 of the ketogenic diet intervention. The intake of vitamins and serum antioxidative potential were also investigated. RESULTS: Throughout the diet the intakes of magnesium, calcium, iron, phosphorus, and potassium were less than recommended values, but serum levels always remained within the reference range. Nevertheless, the level of calcium decreased significantly (from 2.52 ± 0.10 mmol/L at baseline to 2.36 ± 0.07 mmol/L at week 12, P < 0.001), which could be due to the omission of legumes and reduced dairy intake or because of the high fat intake alone. The levels of phosphate increased concomitantly. Calcium serum levels were negatively associated with ω-6 but not with ω-3 unsaturated fatty acid intake. The intakes of water-soluble vitamins were also too low. However, the antioxidative potential of serum did not change during intervention. CONCLUSIONS: Careful choice of foods that will provide the necessary micronutrients is of utmost importance when consuming ketogenic diet. In the 12 wk study the decreased intakes were not reflected in serum values, but special attention to calcium should be advised if such diet is recommended for longer periods.

Effects of Royal Jelly Administration on Lipid Profile, Satiety, Inflammation, and Antioxidant Capacity in Asymptomatic Overweight Adults.

OBJECTIVES: Obesity and overweight are chronic disorders of multifactorial origin that are characterized by high oxidative status and by chronic activation of macrophages in peripheral tissues. Effective therapeutic approaches to lower inflammation and oxidative stress are currently of general interest. Royal jelly (RJ) is a functional food with a broad range of pharmacological activities, mainly used by healthy individuals or borderline patients to protect themselves against disease onset. The objective of this randomized, double-blind, placebo-controlled trial was to investigate the effects of RJ supplementation on metabolic profile and oxidative and inflammatory parameters in asymptomatic overweight adults, considered at an early stage of developing metabolic syndrome. MATERIAL AND METHODS: The experimental group (n=30) was given RJ and the control group (n=30) was provided with a placebo for eight weeks. Anthropometric, biochemical parameters, biomarkers of oxidative stress, and inflammation were assessed at baseline, after 4 and 8 weeks of the intervention, and after additional 2 weeks of follow up. RESULTS AND CONCLUSION: Compared with the placebo, RJ supplementation demonstrated a statistically significant decrease in total cholesterol (6.7%; p=0.041) and inflammatory marker C-reactive protein (19%; p=0.027), whereas significant increases were observed in anti-inflammatory marker adiponectin (34%; p=0.011), endogenous antioxidants bilirubin (35%; p=0.002) and uric acid (5%; p=0.018), total antioxidant capacity in serum (54%; p=0.005), and leptin (17%; p=0.025). The present study demonstrated positive effects of RJ administration on lipid profile, satiety, inflammation, and antioxidant capacity in overweight adults. Therefore, our study supports the benefits of RJ supplementation for the improvement of human health.

Weight loss, improved physical performance, cognitive function, eating behavior, and metabolic profile in a 12-week ketogenic diet in obese adults.

The ketogenic diet (KD) is being increasingly promoted as a strategy to fight obesity. Although the KD is effective for weight loss and weight control, comprehensive determination of its relationship with biochemical, physiological and psychological changes is still largely unexplored. We hypothesized that a 12-week KD (12KD) would significantly affect body weight, physical performance, cognitive function, eating behaviors, the metabolic and hormonal profile in obese adults, although differently in men and women. In an uncontrolled intervention, 35 sedentary obese adults (13 men, 25 women), aged 37 ±â€¯7 years with a BMI 36.1 ±â€¯5.6 kg/m2 underwent a 12KD between March 2017 and June 2017 at the University of Primorska. The 12KD resulted in decreased appetite, significant weight loss of participants (-18 ±â€¯9 kg men vs. -11 ±â€¯3 kg women; P < .001), decreased emotional and external eating (P < .001 for both), increased body image satisfaction (P < .001) and improved physical performance (P < .001). Biochemically, a significant drop in glucose (P = .026), and a significant increase in LDL-cholesterol (P = .031), CRP (P = .007), and BDNF (P = .035) were observed in the first 2 weeks; then, all listed parameters returned to baseline. On the other hand, a significant reduction in insulin (P < .001) and leptin levels (P < .001), and a significant increase in adiponectin (P = .008) and NPY (P = .009) were detected throughout the duration of the 12KD. Our results show the efficacy of the 12KD on weight loss, physical performance, cognitive function, eating behaviors and metabolic profile. However, the long-term effects of a KD on these outcomes needs to be further studied before general recommendations can be made.

RECQ1 Helicase Silencing Decreases the Tumour Growth Rate of U87 Glioblastoma Cell Xenografts in Zebrafish Embryos.

RECQ1 helicase has multiple roles in DNA replication, including restoration of the replication fork and DNA repair, and plays an important role in tumour progression. Its expression is highly elevated in glioblastoma as compared to healthy brain tissue. We studied the effects of small hairpin RNA (shRNA)-induced silencing of RECQ1 helicase on the increase in cell number and the invasion of U87 glioblastoma cells. RECQ1 silencing reduced the rate of increase in the number of U87 cells by 30%. This corresponded with a 40% reduction of the percentage of cells in the G2 phase of the cell cycle, and an accumulation of cells in the G1 phase. These effects were confirmed in vivo, in the brain of zebrafish ( Daniorerio ) embryos, by implanting DsRed-labelled RECQ1 helicase-silenced and control U87 cells. The growth of resulting tumours was quantified by monitoring the increase in xenograft fluorescence intensity during a three-day period with fluorescence microscopy. The reduced rate of tumour growth, by approximately 30% in RECQ1 helicase-silenced cells, was in line with in vitro measurements of the increase in cell number upon RECQ1 helicase silencing. However, RECQ1 helicase silencing did not affect invasive behaviour of U87 cells in the zebrafish brain. This is the first in vivo confirmation that RECQ1 helicase is a promising molecular target in the treatment of glioblastoma.

Topoisomerase IIß mediates the resistance of glioblastoma stem cells to replication stress-inducing drugs.

BACKGROUND: Glioblastoma stem cells (GSC) have been extensively recognized as a plausible cause of glioblastoma resistance to therapy and recurrence resulting in high glioblastoma mortality. Abnormalities in the DNA repair pathways might be responsible for the inability of the currently used chemotherapeutics to eliminate the (GSC) subpopulation. METHODS: In this work, we compared the expression of sixty DNA repair related genes between primary glioblastoma cell cultures and the glioblastoma enriched stem cell primary cultures. MTT test was used to analyze the effect of selected drugs and immunofluorescence to evaluate the load of DNA damage. RESULTS: We found several differentially expressed genes and we identified topoisomerase IIß (Top2ß) as the gene with highest up-regulation in GSC. Also among the tested cell lines the expression of Top2ß was the highest in NCH421k cells, a well-characterized glioblastoma cell line with all the stemness characteristics. On the other hand, Top2ß expression markedly decreased upon the induction of differentiation by all trans-retinoic acid. Depletion of Top2ß increased the sensitivity of NCH421k cells to replication stress inducing drugs, such as cisplatin, methyl-methanesulfonate, hydrogen peroxide, and temozolomide. Consistently, we found an increased load of DNA damage and increased Chk1 activation upon Top2ß depletion in NCH421k cells. CONCLUSION: We suggest that Top2ß may represent a new target for gene therapy in glioblastoma. In addition, the other genes that we found to be up-regulated in GSC versus glioblastoma primary cells should be further investigated as glioblastoma theranostics.