The significance of calprotectin, CD147, APOA4 and DJ-1 in non-invasive detection of urinary bladder carcinoma.

Aim of the study is to define the diagnostic accuracy of selected urinary protein biomarkers in the non-invasive detection of primary and recurrent urothelial carcinoma of the urinary bladder. The urinary levels of calprotectin, CD147, APOA4 and protein deglycase DJ-1 were examined in 255 individuals, including 60 controls with non-malignant urological disease, 61 patients with a history of urinary bladder cancer with negative cytology and negative cystoscopy and 134 patients with urinary bladder cancer. Urinary concentrations of biomarkers were determined by Enzyme-Linked Immunosorbent Assay (ELISA). During the follow-up of patients with non-muscle invasive bladder cancer (NMIBC), a group of 44 patients with cancer recurrence was compared to the group of 61 patients with a history of NMIBC but with no evidence of disease. Urinary concentrations of the evaluated markers did not reveal any significant difference between these groups. During the primary diagnosis, a group of 90 patients with primary bladder cancer and 60 subjects with benign disease were compared. Urinary levels of CD147 were not significantly higher in patients with tumors. The greatest diagnostic accuracy was observed in APOA4 (sensitivity 55.6, specificity 83.3, AUC 0.75), and lesser in calprotectin (sensitivity 39.4, specificity 87.7, AUC 0.66) and in DJ-1 (sensitivity 61.1, specificity 66.7, AUC 0.64), respectively. Apolipoprotein A4 may be used potentially as a supplemental urinary marker in the diagnosis of primary bladder cancer.

Strain differences in intraluminal thread model of middle cerebral artery occlusion in rats.

Stroke is despite of progressive improvements in treatment and reperfusion strategies one of the most devastating human pathology. However, as quality of acute health care improves and more people survive ischemic attack, healthcare specialists have to solve new challenges to preserve reasonable quality of life to these patients. Thus, novel approaches which prevents comorbidities of stroke and improve quality of life of stroke survivors in general has to be developed and experimentally tested. The aim of the present paper was to establish reliable rat model of middle cerebral occlusion and set of methods allowing selection of animals suitable for long-term experiments. We have compared mortality rates, cerebral blood flow and extension of ischemic lesion induced by intraluminal filament in three widely used outbred rat strains. We have additionally used an animal 18F-DG PET scans to verify its reliability in noninvasive detection of ischemic infarct in acute period (24 h after MCAO) for selecting animals eligible for long survival experiments. Our data clearly indicates that high variability between rat strains might negatively influence stroke induction by intraluminal thread occlusion of middle cerebral artery. Most reliable outbred rat strain in our hands was Sprague-Dawley where maximal reduction of cerebral blood flow and extensive ischemic lesion was observed. Contrary, Wistar rats exhibited higher mortality and Long-Evans rats significantly smaller or no ischemic region in comparison to Sprague-Dawley. Additionally, we have confirmed a positron emission tomography with 18F-fluorodeoxyglucose as suitable method to assess extension of ischemic region in acute period after the experimental arterial occlusion in rats.

Rapid screening test of most frequent BRCA1/BRCA2 pathogenic variants in the NGS era.

The average risk of breast cancer in general Slovak population of women is 4-5% and the risk of ovarian cancer is 2%. Probability of breast/ovarian cancer development is higher in individuals carrying a causative germline DNA variant in BRCA1 or BRCA2 gene responsible for hereditary breast/ovarian cancer (HBOC). Although a major proportion of inherited breast/ovarian cancers are due to the mentioned causal mutations, a number of new genes have emerged. Here we describe a rapid, multiplex and comprehensive approach for the detection of pathogenic variants in BRCA1 and BRCA2 genes which most frequently occur in Slovak HBOC population. Analysis comprises the combination of mutation specific methods. Fluorescent PCR amplification followed by fragment analysis for detection of insertions/deletions in exon 11 of BRCA1 gene. Second method is SNaPshot analysis for detection of the most frequent missense and ins/del variants in exons 2, 5, 13, 20 of BRCA1 and exons 11, 23 and 25 of BRCA2 gene. Altogether, we have analyzed 687 samples, 86 (12.5%) in group 1, which fulfilled indication criteria based on the positive family/personal history. Group 2 involved 601 (87.5%) cases, who did not meet the indication criteria and only the screening test was recommended. Using the combined approach, we have identified 47 mutated samples (6.8%), 40 in group 1 (46.5%) and 7 in group 2 (1.1%). However, the presented screening test would not provide complex results of BRCA1/2 gene analysis, it offers testing accessible to a broader spectrum of individuals under the threshold of indication for whole gene analysis. This approach may provide valuable information even in the NGS analysis era.

Plasma Phospholipid Fatty Acid Profile is Altered in Both Septic and Non-Septic Critically Ill: A Correlation with Inflammatory Markers and Albumin.

This study analyzes fatty acid (FA) composition in plasma lipids and erythrocyte phospholipids while comparing septic and non-septic critically ill patients. The aim was to describe impacts of infection and the inflammatory process. Patients with severe sepsis (SP, n = 13); age-, sex- and APACHE II score-matched non-septic critically ill with systemic inflammatory response syndrome (NSP, n = 13); and age-/sex-matched healthy controls (HC, n = 13) were included in a prospective case-control study during the first 24 h after admission to the intensive care unit. In both SP and NSP, lower n-6 polyunsaturated FA (PUFA) accompanied by higher proportions of monounsaturated FA (MUFA) in plasma phospholipids (PPL) was observed relative to HC. MUFA proportion was negatively correlated with n-6 PUFA, high density lipoprotein cholesterol (HDL-C), and albumin. MUFA was positively correlated with C-reactive protein (CRP), procalcitonin (PCT), interleukins (IL-6, IL-10), oxidized low density lipoproteins (ox-LDL), and conjugated dienes (CD). In both SP and NSP, inflammatory and lipid peroxidation markers were significantly higher-CRP (p < 0.001; p = 0.08), IL-6, IL-10, TNF-α (p < 0.01, p = 0.06), ox-LDL, and CD while total cholesterol, HDL-C, LDL-C albumin, and 20:4n-6/22:6n-3 and n-6/n-3 ratios were lower compared to HC. In conclusion, the changes in plasma lipid FA profile relate to the intensity of inflammatory and peroxidative response regardless of insult etiology. The lower MUFA and higher n-6 PUFA proportions in PPL were inversely correlated with cholesterol and albumin levels.

Could human cold adaptation decrease the risk of cardiovascular disease?

The impact of repeated exposure to cold and cold adaptation on human cardiovascular health is not fully understood. The aim of the study was to determine the effect of cold adaptation on cardiovascular risk factors, thyroid hormones and the capacity of humans to reset the damaging effect of oxidative stress. Ten well cold-adapted winter swimmers (CA) and 16 non-adapted controls (CON) were enroled in this experiment to test whether cold adaptation could influence the parameters of lipoprotein metabolism, cholesterol efflux capacity (CEC), homocysteine, thyroid hormones, antioxidant defence markers (reduced glutathione (GSH), glutathione peroxidase 1 (GPX1), glutathione reductase (GR), catalase (CAT) and paraoxonase 1 (PON1)) and oxidative stress markers (concentration of conjugated dienes (CD)). A decreased apolipoprotein B/apolipoprotein A1 (ApoB/ApoA1) ratio was found in the CA group (p<0.05), but other lipoprotein parameters, including CEC, did not differ significantly. Plasma homocysteine was lower in CA subjects in comparison with controls (p<0.05). Higher triiodothyronine (T3) values were observed in the CA compared to the CON (p<0.05) group, but TSH and other thyroid hormones did not differ between both groups. CA subjects had lower activity of GPX1 (p<0.05), lower concentrations of CD (p<0.05) and increased activities of PON1 (p<0.001) compared to CON subjects. A trend for decreased activity of CAT (p=0.06) in CA compared to CON groups was also observed, but GSH levels did not differ significantly. Zn concentration was higher in the CA group than in the CON group (p<0.001). Human cold adaptation can influence oxidative stress markers. Trends towards the improvement of cardiovascular risk factors in cold-adapted subjects also indicate the positive effect of cold adaptation on cardio-protective mechanisms.

Human catalase, its polymorphisms, regulation and changes of its activity in different diseases.

Catalase (CAT) is a well-studied enzyme that plays an important role in protecting cells against the toxic effects of hydrogen peroxide. In human, it has been implicated in different physiological and pathological conditions. This review summarizes the information available on the function and role of CAT polymorphisms in pathogenesis of various pathophysiological states as well as on the regulation of CAT gene expression. Numerous studies have described the CAT polymorphisms and their link with various diseases. Changes in the CAT levels were reported in many different diseases and polymorphisms in the CAT gene were shown to be associated with different pathophysiological states, e.g. hypertension, diabetes mellitus, insulin resistance, dyslipidaemia, asthma, bone metabolism or vitiligo. Regulation of the CAT gene expression plays an important role in the levels of CAT. The catalase gene expression is regulated by various mechanisms involving e.g. peroxisome proliferator-activated receptor γ (PPARγ), tumour necrosis factor α (TNF-α), p53 protein and hypermethylation of CpG islands in the catalase promoter. Transcription of the CAT gene is mainly influenced by the -262 C/T and -844 A/G polymorphisms. A common polymorphism -262 C/T in the promoter region has been found to be associated with altered CAT activities. Apart from genetic factors, the activities of CAT may be affected by age, seasonal variations, physical activity, or a number of chemical compounds. Future investigations are necessary to elucidate the role of CAT in pathogenesis of oxidative stress-related diseases.

Paraoxonase-1 (PON1) status in pancreatic cancer: relation to clinical parameters.

Human paraoxonase 1 (PON1) has been shown to decrease the level of systemic oxidative stress, which is thought to contribute to cancer development. The aim of this study was to examine the interrelationships between PON1 status and some clinical characteristics in patients with pancreatic cancer (PC). A group of 73 consecutive patients with PC (stage II-IV) and 73 control subjects were examined. Laboratory studies included five polymorphisms of the PON1 gene (L55M, Q192R, -108C/T, -126C/T, and -162A/G), PON1 arylesterase (PON1-A) and lactonase (PON1-L) activities, as well as some markers of protein metabolism, insulin resistance, and oxidative stress. In comparison with the control group, no difference in the distribution of the PON1 polymorphisms was found in cancer patients, both arylesterase and lactonase activities being significantly lower (-33, -47 %, respectively, both P < 0.001). There was neither statistically significant association of PON1 polymorphisms with tumour stages nor with diabetes mellitus connected with PC. The genotype distribution of L55M and ­108C/T differed only in a subgroup of patients presenting clinically relevant malnutrition (χ² = 6.50, 6.25, respectively, both P < 0.05). In the PC group, PON1-A and PON1-L activities correlated with Nutritional Risk Index (r = 0.351, 0.409, respectively, both P < 0.01), PON1-L with mid-arm muscle circumference (r = 0.328, P < 0.05), and PON1-A and PON1-L with serum albumin (r = 0.352, 0.391 respectively, both < 0.01). Our results suggest that PON1 plays an important role in PC, especially in cancer-associated malnutrition.

Identifying governance strategies that effectively support ecosystem services, resource sustainability, and biodiversity.

Conservation scientists, national governments, and international conservation groups seek to devise, and implement, governance strategies that mitigate human impact on the environment. However, few studies to date have systematically investigated the performance of different systems of governance in achieving successful conservation outcomes. Here, we use a newly-developed analytic framework to conduct analyses of a suite of case studies, linking different governance strategies to standardized scores for delivering ecosystem services, achieving sustainable use of natural resources, and conserving biodiversity, at both local and international levels. Our results: (i) confirm the benefits of adaptive management; and (ii) reveal strong associations for the role of leadership. Our work provides a critical step toward implementing empirically justified governance strategies that are capable of improving the management of human-altered environments, with benefits for both biodiversity and people.

[Features of metabolic syndrome in patients with depressive disorder]. / Rysy metabolického syndromu u nemocných s depresivní poruchou.

BACKGROUND: Depressive disorder is a serious illness with a high incidence, proxime accessit after anxiety disorders among the psychiatric diseases. It is accompanied by an increased risk of development of type 2 diabetes mellitus, cardiovascular disease, and by increased all-cause mortality. Recently published data have suggested that factors connected with the insulin resistance are at the background of this association. METHODS AND RESULTS: In this pilot study we have investigated parameters of lipid metabolism and glucose homeostasis in consecutively admitted patients suffering from depressive disorder (DD) (group of 42 people), in 57 patients with the metabolic syndrome (MetS) and in a control group of 49 apparently healthy persons (CON). Depressive patients did not differ from the control group by age or body mass index (BMI) value, but they had statistically significantly higher concentrations of serum insulin, C-peptide, glucose, triglycerides (TG), conjugated dienes in LDL particles (CD-LDL), higher value of microalbuminuria and of insulin resistance (HOMA-IR) index. They simultaneously had significantly lower value of the insulin sensitivity (QUICKI) index. In comparison with the MetS group the depressive patients were characterized by significantly lower both systolic and diastolic blood pressure, BMI , serum TG, apolipoprotein B, uric acid, C-peptide and by higher concentrations of apolipoprotein A-I and HDL-cholesterol. On the contrary, we have not found statistically significant differences between the DD and MetS groups in the concentrations of serum insulin, glucose, HOMA and QUICKI indices, in CD-LDL and MAU. CONCLUSIONS: In this pilot study, we have found in patients with depressive disorder certain features of metabolic syndrome, especially insulin resistance and oxidative stress.

Towards the development of Bacillus subtilis as a cell factory for membrane proteins and protein complexes.

BACKGROUND: The Gram-positive bacterium Bacillus subtilis is an important producer of high quality industrial enzymes and a few eukaryotic proteins. Most of these proteins are secreted into the growth medium, but successful examples of cytoplasmic protein production are also known. Therefore, one may anticipate that the high protein production potential of B. subtilis can be exploited for protein complexes and membrane proteins to facilitate their functional and structural analysis. The high quality of proteins produced with B. subtilis results from the action of cellular quality control systems that efficiently remove misfolded or incompletely synthesized proteins. Paradoxically, cellular quality control systems also represent bottlenecks for the production of various heterologous proteins at significant concentrations. CONCLUSION: While inactivation of quality control systems has the potential to improve protein production yields, this could be achieved at the expense of product quality. Mechanisms underlying degradation of secretory proteins are nowadays well understood and often controllable. It will therefore be a major challenge for future research to identify and modulate quality control systems of B. subtilis that limit the production of high quality protein complexes and membrane proteins, and to enhance those systems that facilitate assembly of these proteins.

[Composition of the nonesterified fatty acids and lipid peroxidation in metabolic syndrome]. / Slození esterifikovaných mastných kyselin a lipoperoxidace u metabolického syndromu.

BACKGROUND: Composition of the nonesterified fatty acids in plasma in metabolic syndrome patients and in other syndromes of insulin resistance is altered. Fatty acid profile in plasma is related to the composition of dietary fat and to the metabolic changes of fatty acids, e.g. to de novo lipogenesis, beta-oxidation and conversion accompanying the oxidative stress. The aim of the work was to study the fatty acid composition in the major plasma lipid classes in relation to the insulin resistance, to some polymorphisms of candidate genes with activity related to insulin resistance, and to the lipoprotein composition and parameters of lipid peroxidation. METHODS AND RESULTS: 95 patients with metabolic syndrome (56 M/39 F) and 195 healthy persons (99 M/96 F) were included into the cohort. Basic clinical data, parameters of glucose homeostasis, lipid concentration in plasma and conjugated diens in LDL were determined. Fatty acids were detected by capillary gas chromatography. Polymorphisms of apolipoprotein E, intestinal isoforms of fatty acid binding protein (Ala54Thr) and y-2 isoforms of peroxisomal activated receptor (Alal2Pro) were analyzed using combination of polymerase chain reaction methods and by the detection of polymorphisms of the restriction fragment length. Persons with metabolic syndrome had higher concentrations of CRP and conjugated diens in LDL. In all lipid classes we proved a decreased concentration of n-6 polyunsaturated fatty acids and an increase of unsaturated fatty acids. From all the acids, the only significant was the decrease of linolic acid concentration and the increase of palmitic and palmitoyl acids. Results showed an increase of delta 9 palmitic acid desaturase activity, delta 6 linolic acid desaturase and elongase activity. Concentration of conjugated diens in LDL inversely correlated with linolic acid. Clinical or laboratory parameters and homozygotic combination of polymorphism studied were not mutually related. CONCLUSIONS: Changes in the profile of fatty acids during the metabolic syndrome results from the elevated lipogenesis and from the higher level of oxidative stress.