Association of Leukocyte, Erythrocyte, and Platelet Counts with Metabolic Syndrome and Its Components in Young Individuals without Overt Signs of Inflammation: A Cross-Sectional Study.

The presence of metabolic syndrome (MetS) increases the risk of developing type 2 diabetes, cardiovascular diseases, and mortality. MetS is associated with increased leukocyte or erythrocyte counts. In 16- to 20-year-old males (n = 1188) and females (n = 1231) without signs of overt inflammation, we studied whether the presence of MetS and its components results in elevated blood cell counts. The leukocyte, erythrocyte, and thrombocyte counts significantly but weakly correlated with the continuous MetS score, MetS components, uric acid, and C-reactive protein levels both in males (r = -0.09 to 0.2; p < 0.01) and females (r = -0.08 to 0.2; p < 0.05). Subjects with MetS had higher leukocyte (males: 6.2 ± 1.3 vs. 6.9 ± 1.2 × 109/L; females 6.6 ± 1.5 vs. 7.5 ± 1.6 × 109/L; p < 0.001), erythrocyte (males: 5.1 ± 0.3 vs. 5.3 ± 0.3 × 1012/L; females: 4.5 ± 0.3 vs. 4.8 ± 0.3 × 1012/L; p < 0.001), and platelet counts (males: 245 ± 48 vs. 261 ± 47 × 109/L; females: 274 ± 56 vs. 288 ± 74 × 109/L; p < 0.05) than those without MetS. With the exception of platelet counts in females, the blood counts increased with the number of manifested MetS components. Phenotypes with the highest average leukocyte, erythrocyte, or platelet counts differed between sexes, and their prevalence was low (males: 0.3% to 3.9%; females: 1.2% to 2.7%). Whether functional changes in blood elements accompany MetS and whether the increase in blood counts within the reference ranges represents a risk for future manifestation of cardiometabolic diseases remain unanswered.

Mitochondrial DNA variability and Covid-19 in the Slovak population.

Recent studies have shown that mitochondria are involved in the pathogenesis of Covid-19. Mitochondria play a role in production of reactive oxygen species and induction of an innate immune response, both important during infections. Common variability of mitochondrial DNA (mtDNA) can affect oxidative phosphorylation and the risk or lethality of cardiovascular, neurodegenerative diseases and sepsis. However, it is unclear whether susceptibility of severe Covid-19 might be affected by mtDNA variation. Thus, we have analyzed mtDNA in a sample of 446 Slovak patients hospitalized due to Covid-19 and a control population group consisting of 1874 individuals. MtDNA variants in the HVRI region have been analyzed and classified into haplogroups at various phylogenetic levels. Binary logistic regression was used to assess the risk of Covid-19. Haplogroups T1, H11, K and variants 16256C > T, 16265A > C, 16293A > G, 16311 T > C and 16399A > G were associated with an increased Covid-19 risk. On contrary, Haplogroup J1, haplogroup clusters H + U5b and T2b + U5b, and the mtDNA variant 16189 T > C were associated with decreased risk of Covid-19. Following the application of the Bonferroni correction, statistical significance was observed exclusively for the cluster of haplogroups H + U5b. Unsurprisingly, the most significant factor contributing to the mortality of patients with Covid-19 is the age of patients. Our findings suggest that mtDNA haplogroups can play a role in Covid-19 pathogenesis, thus potentially useful in identifying susceptibility to its severe form. To confirm these associations, further studies taking into account the nuclear genome or other non-biological influences are needed.

Metabolic Effects of Anti-TNF-α Treatment in Rheumatoid Arthritis.

Rheumatoid arthritis (RA) is associated with high cardiovascular mortality. It is not clear whether the metabolic consequences of chronic inflammation are involved. Biological disease-modifying anti-rheumatic drugs (bDMARDs) are highly efficient in the treatment of inflammation in RA. In this study, we aimed to describe the metabolic effects of anti-TNF-α treatment in RA patients. The clinical status of 16 patients was assessed using disease activity score-28 (DAS28) and C-reactive protein (CRP). Plasma samples were collected before treatment with anti-TNF-α treatment as well as after three and six months of treatment. Markers of lipid and glucose metabolism, as well as renal biomarkers, were assessed using standard biochemistry. ELISA was used for the quantification of insulin, leptin, and adiponectin. Although fasting insulin decreased by 14% at the end of the study, most of the analyzed parameters did not show any statistically or clinically significant dynamics. The exception was total bilirubin and cholesterol, which increased by 53% and 14%, respectively, after six months of treatment with anti-TNF-α treatment. Anti-TNF-α treatment did not induce major metabolic changes despite the strong anti-inflammatory and clinical symptoms of RA. Further studies will show whether longer observations are required for the detection of the metabolic effects of the anti-inflammatory treatment. Additional research is needed to understand the observed effect of bilirubin as an important endogenous antioxidant.

Association of Atherogenic Index of Plasma with Cardiometabolic Risk Factors and Markers in Lean 14-to-20-Year-Old Individuals: A Cross-Sectional Study.

Cardiometabolic risk factors at a young age pose a significant risk for developing atherosclerotic cardiovascular disease in adulthood. Atherogenic dyslipidemia is highly associated with obesity and metabolic syndrome already in young age. It remains unclear whether cardiometabolic risk factors associate with the atherogenic index of plasma (AIP = log (TAG/HDL-C) in lean subjects with low atherogenic risk. As both the AIP and markers of cardiometabolic risk are continuous variables, we expected their association to be linear before the manifestation of obesity and atherogenic dyslipidemia. We analyzed the prevalence of increased atherogenic risk (AIP ≥ 0.11) in 2012 lean 14-to-20-year-old subjects (55% females) and the trends of cardiometabolic risk factors across the quartiles (Q) of AIP in a subgroup of 1947 (56% females) subjects with low atherogenic risk (AIP < 0.11). The prevalence of AIP ≥ 0.11 reached 3.6% in females and 8.5% in males. HDL-C, non-HDL-C, triglycerides, and the continuous metabolic syndrome score showed a stepwise worsening across the AIP quartiles in both sexes. Measures of obesity and insulin resistance were worse in Q4 vs. Q1 groups, and leukocyte counts were higher in Q4 and Q3 vs. Q1. Females in Q4 presented with a higher C-reactive protein and lower adiponectin, estradiol, and testosterone levels. The multivariate regression model selected non-HDL-C, QUICKI, and erythrocyte counts as significant predictors of AIP in males; and non-HDL-C and C-reactive protein in females. A question arises whether the lean individuals on the upper edge of low atherogenic risk are prone to earlier manifestation of metabolic syndrome and shift to the higher AIP risk group.

Association of Inflammatory and Oxidative Status Markers with Metabolic Syndrome and Its Components in 40-to-45-Year-Old Females: A Cross-Sectional Study.

Oxidative stress and sterile inflammation play roles in the induction and maintenance of metabolic syndrome (MetS). This study cohort included 170 females aged 40 to 45 years who were categorized according to the presentation of MetS components (e.g., central obesity, insulin resistance, atherogenic dyslipidemia, and elevated systolic blood pressure) as controls not presenting a single component (n = 43), those with pre-MetS displaying one to two components (n = 70), and females manifesting MetS, e.g., ≥3 components (n = 53). We analyzed the trends of seventeen oxidative and nine inflammatory status markers across three clinical categories. A multivariate regression of selected oxidative status and inflammatory markers on the components of MetS was performed. Markers of oxidative damage (malondialdehyde and advanced-glycation-end-products-associated fluorescence of plasma) were similar across the groups. Healthy controls displayed lower uricemia and higher bilirubinemia than females with MetS; and lower leukocyte counts, concentrations of C-reactive protein, interleukine-6, and higher levels of carotenoids/lipids and soluble receptors for advanced glycation end-products than those with pre-MetS and MetS. In multivariate regression models, levels of C-reactive protein, uric acid, and interleukine-6 were consistently associated with MetS components, although the impacts of single markers differed. Our data suggest that a proinflammatory imbalance precedes the manifestation of MetS, while an imbalance of oxidative status accompanies overt MetS. Further studies are needed to elucidate whether determining markers beyond traditional ones could help improve the prognosis of subjects at an early stage of MetS.

Severe gestational diabetes mellitus in lean dams is associated with low IL-1α levels and affects the growth of the juvenile mouse offspring.

We investigated how maternal gestational diabetes (GDM) impacts the metabolic status of offspring. GDM was induced in CD1 mice consuming a fast-food diet (FFD) by repeated low-dose streptozotocin injections before mating. Offspring of normoglycemic standard chow or the FFD consuming dams served as controls. In 4-week-old offspring weaned to standard chow, plasma concentrations of extracellular DNA, inflammatory markers, and parameters of the cardiometabolic status (glycemia, liver lipid content; body, organ, and fat weight) were determined. Two-factor analysis of variance indicated that the male offspring of GDM dams manifest postnatal growth retardation and lower relative kidney weight. Regardless of sex, GDM offspring manifest the lowest IL-1α levels, and other inflammatory markers showed mild and inconsistent alterations. Offspring of dams consuming the FFD displayed higher liver triacylglycerols content. The three groups of offspring showed no significant differences in glycemia and extracellular DNA. Partial least squares-discriminant analysis indicated that male GDM offspring present lower kidney, body, and brown adipose tissue weights; lower IL-1α levels, and higher concentrations of GM-CSF and IL-10 compared with their FFD counterparts. The model failed to select discriminative variables in females. In conclusion, in mice, maternal GDM in the absence of obesity adversely affects the early growth of juvenile male offspring.

Association of α-Dicarbonyls and Advanced Glycation End Products with Insulin Resistance in Non-Diabetic Young Subjects: A Case-Control Study.

α-Dicarbonyls and advanced glycation end products (AGEs) may contribute to the pathogenesis of insulin resistance by a variety of mechanisms. To investigate whether young insulin-resistant subjects present markers of increased dicarbonyl stress, we determined serum α-dicarbonyls-methylglyoxal, glyoxal, 3-deoxyglucosone; their derived free- and protein-bound, and urinary AGEs using the UPLC/MS-MS method; soluble receptors for AGEs (sRAGE), and cardiometabolic risk markers in 142 (49% females) insulin resistant (Quantitative Insulin Sensitivity Check Index (QUICKI) ≤ 0.319) and 167 (47% females) age-, and waist-to-height ratio-matched insulin-sensitive controls aged 16-to-22 years. The between-group comparison was performed using the two-factor (sex, presence/absence of insulin resistance) analysis of variance; multiple regression via the orthogonal projection to latent structures model. In comparison with their insulin-sensitive peers, young healthy insulin-resistant individuals without diabetes manifest alterations throughout the α-dicarbonyls-AGEs-sRAGE axis, dominated by higher 3-deoxyglucosone levels. Variables of α-dicarbonyls-AGEs-sRAGE axis were associated with insulin sensitivity independently from cardiometabolic risk markers, and sex-specifically. Cleaved RAGE associates with QUICKI only in males; while multiple α-dicarbonyls and AGEs independently associate with QUICKI particularly in females, who displayed a more advantageous cardiometabolic profile compared with males. Further studies are needed to elucidate whether interventions alleviating dicarbonyl stress ameliorate insulin resistance.

Extracellular DNA concentrations in various aetiologies of acute kidney injury.

Extracellular DNA (ecDNA) in plasma is a non-specific biomarker of tissue damage. Urinary ecDNA, especially of mitochondrial origin, is a potential non-invasive biomarker of kidney damage. Despite prominent tissue damage, ecDNA has not yet been comprehensively analysed in acute kidney injury (AKI). We analysed different fractions of ecDNA, i.e. total, nuclear and mitochondrial, in plasma and urine of children, and different animal models of AKI. We also analysed the activity of the deoxyribonuclease (DNase), which is contributes to the degradation of ecDNA. Patients with AKI had higher total and nuclear ecDNA in both, plasma and urine (sixfold and 12-fold in plasma, and 800-fold in urine, respectively), with no difference in mitochondrial ecDNA. This was mainly found for patients with AKI due to tubulointerstitial nephritis and atypical haemolytic uremic syndrome. Increased plasma ecDNA was also found in animal models of AKI, including adenine nephropathy (fivefold), haemolytic uremic syndrome (fourfold), and ischemia-reperfusion injury (1.5-fold). Total urinary ecDNA was higher in adenine nephropathy and ischemia-reperfusion injury (1300-fold and twofold, respectively). DNase activity in urine was significantly lower in all animal models of AKI in comparison to controls. In conclusion, plasma total and nuclear ecDNA and urinary total ecDNA is increased in patients and animals with particular entities of AKI, suggesting a mechanism-dependent release of ecDNA during AKI. Further studies should focus on the dynamics of ecDNA and its potential role in the pathogenesis of AKI.

The Presence of Hyperhomocysteinemia Does Not Aggravate the Cardiometabolic Risk Imposed by Hyperuricemia in Young Individuals: A Retrospective Analysis of a Cross-Sectional Study.

BACKGROUND: Little research has been conducted into the effects of the combined manifestation of hyperuricemia and hyperhomocysteinemia on cardiometabolic risk factors and markers in young subjects. METHODS: 1298 males and 1402 females, 14-to-20-year-olds, were classified into four groups: 1/normouricemic/normohomocysteinemic, 2/normouricemic/hyperhormohomocysteinemic, 3/hyperuricemic/normohomocysteinemic, and 4/hyperuricemic/hyperhomocysteinemic. Anthropometric measures, blood pressure, plasma glucose, insulin, lipids, markers of renal function, C-reactive protein, asymmetric dimethylarginine, and blood counts were determined. RESULTS: Hyperuricemic males (but not females) had higher odds for hyperhomocysteinemia than normouricemic ones (OR: 1.8; 95% CI: 1.4-2.3; p < 0.001). Homocysteine and uric acid levels correlated directly (males: r = 0.076, females: r = 0.120; p < 0.01, both). Two-factor analysis of variance did not reveal a significant impact of hyperhomocysteinemia on any of the investigated cardiometabolic variables in females; in males, hyperuricemia and hyperhomocysteinemia showed a synergic effect on asymmetric dimethylarginine levels. Among four groups, subjects concurrently manifesting hyperuricemia and hyperhomocysteinemia did not presented the highest continuous metabolic syndrome score-a proxy measure of cardiometabolic risk; neither the multivariate regression model indicated a concurrent significant effect of uric acid and homocysteine on continuous metabolic syndrome score in either sex. CONCLUSION: In young healthy subjects, hyperhomocysteinemia does not aggravate the negative health effects imposed by hyperuricemia.

The Bright Side of Skin Autofluorescence Determination in Children and Adolescents with Newly Diagnosed Type 1 Diabetes Mellitus: A Potential Predictor of Remission?

Skin autofluorescence (SAF) is a noninvasive method reflecting tissue accumulation of advanced glycation end products (AGEs). We investigated whether, in newly diagnosed children and adolescents with type 1 diabetes (T1D), this surrogate marker of long-term glycemia is associated with markers of the early manifestation phase, residual secretion capacity of the ß-cells, and the occurrence of remission. SAF was measured in 114 children and adolescents (age: 8.0 ± 4.5 years, 44% girls) at the time of T1D diagnosis, and related to HbA1c, C-peptide, diabetic ketoacidosis, and remission. 56 patients were followed up for 1 year. Seventy-four sex- and age-matched healthy individuals served as controls. SAF was higher in the T1D group compared with controls (1.0 ± 0.2 vs. 0.9 ± 0.2, p < 0.001). At the time of diagnosis, SAF correlated with HbA1c (r = 0.285, p = 0.002), was similar in patients with and without ketoacidosis, and was lower in the remitters compared with non-remitters (0.95 ± 0.18 vs. 1.04 ± 0.26, p = 0.027). Unlike HbA1c, SAF was an independent predictor of remission (∆R2 = 0.051, p = 0.004). Former studies consider SAF in diabetic patients as a tool to identify individuals at an increased risk of chronic complications. Here we show that determination of SAF at the time of T1D diagnosis might potentially predict remission, at least in children.

Long-Term Consumption of a Sugar-Sweetened Soft Drink in Combination with a Western-Type Diet Is Associated with Morphological and Molecular Changes of Taste Markers Independent of Body Weight Development in Mice.

We investigated whether the long-term intake of a typical sugar-sweetened soft drink (sugar-sweetened beverage, SSB) alters markers for taste function when combined with a standard diet (chow) or a model chow mimicking a Western diet (WD). Adult male CD1 mice had ad libitum access to tap water or SSB in combination with either the chow or the WD for 24 weeks. Energy intake from fluid and food was monitored three times a week. Cardiometabolic markers (body weight and composition, waist circumference, glucose and lipid profile, and blood pressure) were analyzed at the end of the intervention, as was the number and size of the fungiform papillae as well as mRNA levels of genes associated with the different cell types of taste buds and taste receptors in the circumvallate papillae using a cDNA microarray and qPCR. Although the overall energy intake was higher in the WD groups, there was no difference in body weight or other cardiometabolic markers between the SSB and water groups. The chemosensory surface from the fungiform papillae was reduced by 36 ± 19% (p < 0.05) in the WD group after SSB compared to water intake. In conclusion, the consumption of the SSB reduced the chemosensory surface of the fungiform papillae of CD1 mice when applied in combination with a WD independent of body weight. The data suggest synergistic effects of a high sugar-high fat diet on taste dysfunction, which could further influence food intake and promote a vicious cycle of overeating and taste dysfunction.

Lean insulin-resistant young adults display increased cardiometabolic risk: A retrospective cross-sectional study.

AIM: We investigated whether lean insulin-resistant individuals manifest increased cardiometabolic risk. METHODS: 2,341 (51.8% females) healthy 16-23-year-old subjects were categorized as lean or overweight/obese; and insulin-sensitive or insulin-resistant, and compared. RESULTS: In both sexes, lean insulin-sensitive and insulin-resistant subjects displayed similar measures of obesity (e.g., males, waist-to-height ratio: lean insulin-sensitive: 0.42 ± 0.03, lean insulin-resistant: 0.43 ± 0.03, overweight/obese insulin-sensitive: 0.49 ± 0.05, overweight/obese insulin-resistant: 0.53 ± 0.06). Lean insulin-sensitive individuals were more insulin-sensitive compared with their overweight/obese peers; insulin-resistant groups presented similar insulin-sensitivity (males, the Quantitative insulin-sensitivity check index (QUICKI): lean insulin-sensitive: 0.354 ± 0.022, lean insulin-resistant: 0.304 ± 0.013, overweight/obese insulin-sensitive: 0.343 ± 0.019, overweight/obese insulin-resistant: 0.299 ± 0.015). The two-factor analysis of variance indicated an independent effect of insulin sensitivity, overweight/obesity, and their interaction on the continuous metabolic syndrome score (p < 0.001, all; males, lean insulin-sensitive: 1.87 ± 0.35, lean insulin-resistant: 2.14 ± 0.42, overweight/obese insulin-sensitive: 2.15 ± 0.40, overweight/obese insulin-resistant: 2.75 ± 0.69). C-reactive protein, leukocyte count, and glomerular filtration rate in both sexes; uric acid, asymmetric dimethyl-arginine, and soluble vascular adhesion protein-1 in males; and soluble receptor for advanced glycation end-products in females were independently associated with insulin resistance. Among phenotypes associated with low QUICKI, the distribution of insulin-resistant individuals was random. CONCLUSION: Later clinical consequences of insulin resistance in lean subjects remain to be elucidated in longitudinal studies.

Bone mineral density and oxidative stress in adolescent girls with anorexia nervosa.

Oxidative stress appears to be involved in the pathogenesis of osteoporosis-a serious complication of anorexia nervosa (AN). We evaluated the oxidative status in adolescent girls with AN and its potential relationship with bone mineral density (BMD). Girls with AN (n = 43) and age-matched healthy controls (n = 20) underwent anthropometric and BMD examination. Markers of bone turnover, oxidative stress, and antioxidant status were measured. Participants with AN and controls did not differ in BMD at the lumbar spine (p = 0.17) and total body less head BMD (p = 0.08). BMD at the total hip was lower (p < 0.001) in the AN group compared with the controls. Levels of antioxidant status markers-ferric reduction antioxidant power, total antioxidant capacity, and reduced and oxidized glutathione ratio (all p < 0.001)-were significantly lower, whereas those of advanced oxidation protein products (AOPP), fructosamines, and advanced glycation end products (AGEs) (all p < 0.001) were higher in AN patients than in healthy controls. BMD and bone turnover markers were positively correlated with antioxidant status markers, while they were negatively correlated with AOPP, fructosamines, and AGEs levels.  Conclusion: This is the first study to assess a potential association between oxidative status and BMD in adolescents with AN. We demonstrated that in young girls, the imbalance of oxidative status and reduced BMD are concurrently manifested at the time of the diagnosis of AN. Disturbance of oxidative status could play a pathogenetic role in AN-associated decreased BMD. What is Known: • Osteoporosis is a serious complication of AN, and in affected adolescents may result in a permanent deficit in bone mass. • Oxidative and carbonyl stress may be involved in the development of bone loss. What is New: • Adolescents girls with AN have impaired antioxidant defense and increased oxidative damage to biomolecules. • Disturbance of oxidative status could affect bone loss and could contribute to decreased BMD in adolescent females with AN.

Creatinine-Based Formulae Poorly Match in the Classification of Hypofiltration or Hyperfiltration in a General Population of Adolescents: A Retrospective Analysis of a Cross-Sectional Study.

Pediatric formulae to estimate glomerular filtration rate (eGFR) give a broad range of values. Their consistency in assigning the subjects as hypofiltrating or hyperfiltrating is unknown. In 1993 apparently healthy adolescents (53.4% females) aged 14-17 years, we investigated the concordance of six creatinine-based formulae in the classification of the subjects into ≤ 5th or ≥95th percentile of eGFR, and the between-groups difference in the prevalence of cardiometabolic risk factors. Mean eGFR varied between 77 and 121 mL/min/1.73 m2. Arbitrary setting of hypofiltration or hyperfiltration to 5% returned 46 males and 53 females. At least one formula classified 89 males and 99 females as hypofiltrating and 105 males and 114 females as hyperfiltrating. All six formulae concordantly classified 15 males and 17 females as hypofiltrating and 9 and 14, respectively, as hyperfiltrating. Pairwise, formulae consistently classified hypofiltration in 42-87% of subjects with hyperfiltration in 28-94%. According to two out of the six formulae, hyperfiltration was associated with an increased prevalence of obesity and obesity-associated comorbidities. Hypofiltrating subjects did not manifest chronic kidney disease-associated comorbidities. Further studies in different populations of healthy adolescents are needed before it is possible to conclude which creatinine-based formula is appropriate for the classification of hypofiltration and hyperfiltration in nonclinical cohorts.

Circulating extracellular DNA is in association with continuous metabolic syndrome score in healthy adolescents.

Obesity is associated with chronic low-grade inflammation that eventually leads to metabolic complications. Extracellular DNA (ecDNA) is a damage-associated molecular pattern. Extracellular mitochondrial DNA can activate innate immunity. We hypothesized that ecDNA, especially of mitochondrial origin, could be associated with components of the metabolic syndrome in young healthy probands. In a cross-sectional study, healthy adolescents (n = 1,249) provided blood samples. Anthropometric data, blood pressure, and blood counts were assessed. In addition, biochemical analysis of sera or plasma was conducted, including the quantification of advanced oxidation protein products (AOPPs) as a marker of oxidative stress induced by neutrophil or monocyte activation. Plasma ecDNA was isolated and measured by fluorometry. Nuclear and mitochondrial DNA were quantified by real-time PCR. Males had higher total plasma ecDNA [15 (11-21) vs. 11 (8-17) ng/mL; median (interquartile range)], nuclear [1,760 (956-3,273) vs. 1,153 (600-2,292) genome equivalents (GE)/mL], and mitochondrial [37,181 (14,836-90,896) vs. 30,089 (12,587-72,286) GE/mL] DNA. ecDNA correlated positively with the continuous metabolic syndrome score (r = 0.158 for males and r = 0.134 for females). Stronger correlations were found between ecDNA of mitochondrial origin and AOPP (r = 0.202 and 0.186 for males and females, respectively). Multivariate regression analysis revealed associations of nuclear DNA with leukocyte and erythrocyte counts. The results of this study of healthy adolescents show that circulating ecDNA is associated with the risk of metabolic syndrome, not with obesity per se. The association between mitochondrial ecDNA and AOPP requires further attention as it supports a potential role of mitochondria-induced sterile inflammation in the pathogenesis of the metabolic syndrome.

A considerable proportion of metabolic syndrome-free adults from Bratislava Region, Slovakia, display an increased cardiometabolic burden.

Although the dichotomous classification of metabolic syndrome (MS) enables the classification of individuals as MS-free or presenting MS, it is inconvenient for assessing cardiometabolic risk in MS-free individuals. Continuous MS score allows for estimation of cardiometabolic burden even in MS-free subjects. We used the scores to estimate the proportion of MS-free subjects on high cardiometabolic risk. A total of 876 subjects (62% females) of Central European descent, aged 20-81 years, were included. International Diabetes Federation (IDF) criteria were employed to classify MS. Continuous scores were calculated. We used the receiver operating characteristics (ROC) analysis to estimate the cutoff value to determine the proportion of MS-free subjects on increased risk. Using the waist circumference, 38% of males and 23% of females presented MS. ROC area under the curves (90%-98%) showed an acceptable performance of both scores to classify the presence of MS. Up to 18% of MS-free males and up to 10% of females displayed continuous score ≥ the relevant cutoff point. The waist-to-height ratio performed similar results. Both continuous scores were proven credible for assessing cardiometabolic risk in MS-free subjects. Clinically, this is important for earlier intervention. Despite minor differences between waist circumference and waist-to-height ratio, it would be appropriate to objectify it using reference population. Novelty: The first study using Z-MSS/siMSS (population-specific Z-score/continuous score of MS) to estimate cardiometabolic risk in Slovak adults. A proportion of MS-free Slovak adults is on high cardiometabolic risk. Difference between using waist circumference and the waist-to-height ratio does not seem to be major in our cohort.

Oxidative status in plasma, urine and saliva of girls with anorexia nervosa and healthy controls: a cross-sectional study.

BACKGROUND: Anorexia nervosa (AN) is a serious psychosomatic disorder with unclear pathomechanisms. Metabolic dysregulation is associated with disruption of redox homeostasis that might play a pivotal role in the development of AN. The aim of our study was to assess oxidative status and carbonyl stress in plasma, urine and saliva of patients with AN and healthy controls. METHODS: Plasma, spot urine, and saliva were collected from 111 girls with AN (aged from 10 to 18 years) and from 29 age-matched controls. Markers of oxidative stress and antioxidant status were measured using spectrophotometric and fluorometric methods. RESULTS: Plasma advanced oxidation protein products (AOPP) and advanced glycation end products (AGEs) were significantly higher in patients with AN than in healthy controls (by 96, and 82%, respectively). Accordingly, urinary concentrations of AOPP and fructosamines and salivary concentrations of AGEs were higher in girls with AN compared with controls (by 250, and 41% in urine; by 92% in saliva, respectively). Concentrations of thiobarbituric acid reactive substances (TBARS) in saliva were 3-times higher in the patients with AN than in the controls. Overall antioxidants were lower in plasma of girls with AN compared to the controls, as shown by total antioxidant capacity and ratio of reduced and oxidized glutathione (by 43, and 31%, respectively). CONCLUSIONS: This is the first study assessing wide range of markers of oxidative status in plasma, urine and saliva of the patients with AN. We showed that both, higher levels of markers of oxidative stress and lower antioxidants play a role in redox disruption. Restoration of redox homeostasis might be of the clinical relevance.

Neurodevelopmental testing of mice in the neonatal period does not affect their locomotor activity, depressive- and anxiety-like behaviour in adolescence.

Neonatal life is a sensitive period of brain plasticity. There are concerns that pre-weaning handling may therefore alter behavioural phenotype in adolescence or adulthood. Since neurodevelopment tests require daily manipulation with pups, later behavioural outcomes might be biased by repeated handling during suckling period. The aim of our study was to assess whether daily neurodevelopmental testing would cause alterations in behavioural phenotype. Sixty-four CD1 mice were randomized to tested and not tested group. In the tested group, maturation of physical features and neurodevelopment were monitored daily from postnatal day 1-21 between 9 and 11 AM. After weaning, battery of behavioural tests to monitor anxiety-like, depressive, or repetitive behaviour was performed in all mice. We revealed no significant between-group differences in performance of these tests. Our data did not confirm the assumption that early neurodevelopment testing during suckling period affects behavioural phenotype in adolescence.

Elevated blood pressure-associated cardiometabolic risk factors and biomarkers in 16-23 years old students with or without metabolic abnormalities.

In obesity, cardiometabolic risk markers show worsening trends with increasing blood pressure (BP). We assumed that risk markers show similar trends across BP categories (normotension, high normal BP, hypertension) in metabolic abnormalities-free subjects (without obesity, insulin resistance, atherogenic dyslipidemia, hyperuricemia, microinflammation) and those presenting them. Data from 2547 (48.1% males) subjects aged 16-23 years were analyzed. The prevalence of males increased across BP categories. Forty-seven percent of individuals with elevated BP were metabolic abnormalities-free. Among 1461 metabolic abnormalities-free subjects, 9% had high normal BP, and 4% hypertension; among 1086 individuals presenting metabolic abnormalities, the prevalence reached 13% and 6%, respectively, (p < 0.001). Both groups displayed similar BP values in corresponding BP categories and significant trends in markers of adiposity, insulin resistance, HDL-cholesterol, atherogenic index of plasma, uric acid, adiponectinemia, and antioxidant capacity of plasma across BP categories. In metabolic abnormalities-free individuals, also significant trends in soluble receptors for advanced glycation end products were revealed. Continuous metabolic syndrome score, a measure of cardiometabolic risk, increased across BP categories regardless of presence or absence of metabolic abnormalities. Multivariate regression models selected male gender, fat-free mass, and uric acid as significant independent predictors for determining BP. Our data emphasize that having a BP outside the normal range significantly worsens risk for cardiometabolic disease in young individuals even if the thresholds for any of the risk factors are not exceeded. Longitudinal studies are needed to assess whether in patients with elevated BP the prognosis of adverse outcomes differs between those presenting and not presenting metabolic abnormalities.

Dynamics of salivary markers of kidney functions in acute and chronic kidney diseases.

Saliva can be used as an alternative diagnostic fluid enabling easy and non-invasive disease monitoring. Urea and creatinine can be measured in saliva and both were shown to be increased in renal failure. However, the dynamics of these markers during the development of kidney diseases is unknown. We aimed to describe the dynamics of salivary urea and creatinine in various animal models of acute kidney injury (AKI) and chronic kidney disease (CKD) and in patients with different stages AKI or CKD. Ninety Wistar rats underwent bilateral nephrectomy (BNX), ischemia-reperfusion injury (IRI) or glycerol-induced kidney injury to model AKI. CKD was modelled using 5/6 nephrectomy. In the clinical part 57 children aged 12.6 ± 4.9 years with AKI (n = 11) or CKD (n = 46) and 29 healthy controls (aged 10.2 ± 3.7 years) were enrolled. Saliva and blood samples were collected in both, animal experiments and the human study. In animal models of AKI, plasma urea and creatinine were higher than in controls. An increase of salivary urea and creatinine (twofold) was observed in BNX and IRI, but only after 12 h and 24 h, respectively. In glycerol nephropathy and 5/6 nephrectomy, salivary urea increased (by 100% and by 50%), while salivary creatinine did not change during the observation period. Salivary urea and creatinine were significantly higher in all patients compared to controls (threefold) and in both, AKI and CKD they were associated with the severity of renal failure. Plasma and salivary concentrations correlated only in children with renal failure (R = 0.72 for urea; R = 0.93 for creatinine), but not in controls (R = -0.007 for urea; R = 0.02 for creatinine). Our study indicates that during the development of renal impairment saliva could be used for non-invasive monitoring in higher stages of AKI or CKD, rather than for screening of early stages of kidney diseases.