The value of human epididymis protein 4 (HE4) as a serum tumor marker for accurate bone metastases finding by whole-body bone scintigraphy in lung cancer patients.

The aim of our study was to correlate the serum concentration of human epididymis protein 4 (HE4) in lung cancer patients with the bone metastases detected by whole-body bone scintigraphy. The serum concentrations of HE4 were determined by electrochemiluminescence immunoassay method in 60 patients with lung cancer and in 10 persons without malignant disease (control group). All participants were examined by whole-body bone scintigraphy with hybrid gamma camera of type BrightView XCT. We found bone metastases in 25.0% of patients by whole-body bone scintigraphy and probable bone metastases in 18.3% of patients. We did not observe bone metastases in 56.7% of patients and in nobody from control group. We observed that 73.33% patients with bone metastases had more than 3 bone metastasis deposits. Patients had significantly increased concentration of HE4 (p < 0.0001). All three subgroups of patients (bone metastases, probable bone metastases, no evidence of bone metastases) had significantly increased concentration of HE4 compared to controls. The highest concentration of HE4 had 9 patients with small-cell lung cancer of whose 4 patients had bone metastases, 4 patients had probable bone metastases and one patient was with no evidence of bone metastases. We found that HE4 has a discriminatory ability to differentiate groups of patients and healthy controls, as well as within scaffold scintigraphy in patients with lung cancer (p = 0.0002). The serum concentration of human epididymis protein 4 was significantly increased in patients with lung cancer in comparison with persons of control group. A quarter of lung cancer patients had identified bone metastases by whole-body bone scintigraphy and approximately 20% of patients had probable bone metastases. The increasing serum concentrations of human epididymis protein 4 can have importance in the diagnosis of bone metastases in patients with lung cancer, in particular in small-cell lung cancer.

Association between interleukin-18 variants and prostate cancer in Slovak population.

Interleukin-18 (IL-18), pro-inflammatory cytokine, plays important role in antitumor immunity. Polymorphisms in the IL-18 gene may lead to its altered production/activity and such modulate susceptibility to prostate cancer. The aim of this study was to evaluate the relationship between the -607 and +105 polymorphisms in the IL-18 gene and the risk of prostate cancer development and progression in Slovak population. The study was performed using 425 patients with prostate cancer, 270 patients with benign prostatic hyperplasia (BHP) and 263 healthy male controls. The statistically significant association of the -607 AC genotype (OR = 2.24; p < 0.001), CC genotype (OR = 1.86; p = 0.006), as well as C allele (OR = 1.27; p = 0.033) with the higher risk of prostate cancer development was observed. No association of the IL-18 -607 polymorphism and BHP was detected. The subset analysis revealed the significant association of the -607 AC genotype (OR = 2.01; p = 0.008) with development of higher-grade carcinomas (Gleason score ≥7) and the strong association of the -607 AC genotype (OR = 3.11; p < 0.001), CC genotype (OR = 2.96; p < 0.001) as well as C allele (OR = 1.51; p = 0.003) with the higher risk of prostate cancer development in the group of patients with PSA < 10 ng/ml. The -607 AC genotype was also connected with significantly higher IL-18 plasma concentrations. No association between the IL-18 +105 polymorphism and prostate cancer was observed. The analysis of the distribution of the -607 and +105 haplotypes showed significant association of the - 607 C/ + 105 A and - 607 C/ + 105 C haplotypes with the risk of prostate cancer. This study found that the IL-18 -607 promoter polymorphism could contribute to prostate cancer development in Slovak population. Its presence was also associated with development of higher-grade carcinomas and therefore may influences the prognosis and aggressiveness of the disease.

[Factors modifying cord blood IgE levels - a pilot study]. / Faktory modifikujúce koncentráciu IgE v pupocníkovej krvi - pilotná stúdia.

AIM: The complex influence of internal and external environmental factors on the individual and his/her immune system and the lack of suitable markers to assess and reduce the risk of the development of allergies during the lifetime can explain the continuous increase in the number of people affected by some form of allergy. According to the results of some studies, cord blood IgE level could be a useful early marker for assessing the risk of atopic diseases, but the studies showed controversial results. In addition, several authors discuss the origin of these antibodies (synthesis in utero, peripartum contamination from maternal blood or placental transfer). The aim of our pilot study was to investigate the possible influence of modifying factors on cord blood IgE level. MATERIAL AND METHODS: Our group of patients consisted of 184 retrospectively selected neonates (98 boys, 53.3% and 86 girls, 46.7%) from whom cord blood was collected and cord blood IgE level was measured 25 years ago (PRIST method). The impact of selected modifying factors (sex, type of delivery or month of birth) on cord blood IgE level was assessed retrospectively. RESULTS: Higher cord blood IgE levels were found in boys than in girls, in neonates born by Caesarean section than in those born by natural delivery, and in those born in the winter months than in other seasons of the year. Our findings are in agreement with those of other authors. CONCLUSION: Based on our results and those of others, we assume that the selected factors affect the cord blood IgE levels to varying degrees. These facts should be taken into consideration while interpreting the cord blood IgE levels.

[The possibility of the serum concentration of osteocalcin determination in lung cancer patients with suspected bone metastases]. / Moznosti stanovenia sérovej koncentrácie osteokalcínu u pacientov s karcinómom plúc pri podozrení na kostné metastázy.

AIM: The aim of this study was to compare serum levels of biochemical markers of bone metabolism - osteocalcin (OC) in correlation with bone metastases found during whole-body bone scintigraphy in patients with lung cancer. MATERIAL AND METHODS: The serum levels of OC as a bone formation marker were determined in 60 patients (46 male, 14 female, mean age 66.65, range 50-84 years) with lung cancer (51 non-small-cell lung cancers - NSCLC and nine small-cell lung cancers - SCLC) and correlated with the presence of bone metastases detected by whole-body bone scintigraphy (hybrid system SPECT/CT: BrightView XCT, Philips Healthcare). Whole-body bone scintigraphy results were compared with OC for each patient with lung cancer and for person from control group of 10 persons (two males, eight females, mean age 52.3, range 34-67 years) with-out malignant disease. RESULTS: By whole-body bone scintigraphy, bone metastases were found in 15 cases (25%), probably in 11 cases (18.33%) and 34 patients (56.67%) were without bone metastases out of 60 patients with lung cancer. The serum levels of OC were above reference range in five cases (8.33%) only with NSCLC and below reference range in 12 cases (20%) - in 10 cases in patients with NSCLC and in two cases in patients with SCLC. In control group of 10 persons, serum level of OC was below reference range only in one case. CONCLUSION: The serum concentrations of osteocalcin were not correlated with findings performed bywhole-body bone scintigraphy in patients with lung cancer. Osteocalcin serum levels determination probably does not have diagnostic importance in lung cancer patients with suspected bone metastases.

Age-associated changes in Ca(2+)-ATPase and oxidative damage in sarcoplasmic reticulum of rat heart.

Altered Ca(2+) handling may be responsible for the development of cardiac contractile dysfunctions with advanced age. In the present study, we investigated the roles of oxidative damage to sarcoplasmic reticulum (SR) and expression of Ca(2+)-ATPase (SERCA 2a) and phospholamban in age-associated dysfunction of cardiac SR. SR vesicles were prepared from hearts of 2-, 6-, 15-, and 26-month-old Wistar rats. Although activity of Ca(2+)-ATPase decreased with advancing age, no differences in relative amounts of SERCA 2a and phospholamban protein were observed. On the other hand, significant accumulation of protein oxidative damage occurred with aging. The results of this study suggest that age-related alteration in Ca(2+)-ATPase activity in the rat heart is not a consequence of decreased protein levels of SERCA 2a and phospholamban, but could arise from oxidative modifications of SR proteins. Cellular oxidative damage caused by reactive oxygen species could contribute to age-related alternations in myocardial relaxation.

Association of oxidative stress and GST-T1 gene with childhood bronchial asthma.

There is increasing evidence that bronchial asthma is associated with increased oxidative stress. Reactive oxygen species are produced in each organism as by-products of several essential reactions and can damage biomolecules. Asthma is a complex chronic inflammatory disorder of the airways, with many candidate genes suspected as being important in its development. Glutathione S-transferase (GST) is such a gene due to its role in protection against oxidative stress. In the present study, we examined the hypothesis that increased oxidative stress and polymorphism in the GST-T1 gene are associated with childhood asthma. We found that the amount of sulfhydryl groups significantly decreased and the content of thiobarbituric acid-reactive substances increased in the group of asthmatic children, compared with healthy controls. The GST-T1 null genotype was more frequent among the asthma patients. These results suggest that the GST-T1 null genotype and increased oxidative stress may play a role in the asthma pathogenesis in children.

Cough reflex sensitivity in various phenotypes of childhood asthma.

Cough is a major symptom in some children with asthma, but the relationship between cough and the severity of asthma is defined insufficiently. As cough represents common problem of pediatrics, several objective methods for its assessment were developed. Cough reflex sensitivity (CRS) test with capsaicin is one of the most important tools for studying cough. In the present study, we aimed to study the CRS in various phenotypes of childhood asthma. We found that, in general, CRS was increased in asthmatic children compared with controls. The most evident increase of CRS was observed during acute asthma exacerbation, in children suffering from asthma with concomitant allergic rhinitis, and in atopic asthmatics. Interestingly, we noted a significant decline in lung function after capsaicin CRS. Various laboratory and clinical characteristics of asthmatic children influence cough sensitivity to a different extent. Cough reflex sensitivity measurement can add valuable information beside the commonly used spirometric and inflammometric methods in the management of asthmatic children.

Factors attributable to the level of exhaled nitric oxide in asthmatic children.

BACKGROUND: Asthma is a heterogeneous disease with variable symptoms especially in children. Exhaled nitric oxide (FeNO) has proved to be a marker of inflammation in the airways and has become a substantial part of clinical management of asthmatic children due to its potential to predict possible exacerbation and adjust the dose of inhalant corticosteroids. OBJECTIVES: We analyzed potential factors that contribute to the variability of nitric oxide in various clinical and laboratory conditions. MATERIAL AND METHODS: Study population consisted of 222 asthmatic children and 27 healthy control subjects. All children underwent a panel of tests: fractioned exhaled nitric oxide, exhaled carbon monoxide, asthma control test scoring, blood sampling, skin prick tests, and basic spirometry. RESULTS: FeNO and other investigated parameters widely changed according to clinical or laboratory characteristics of the tested children. Asthmatics showed increased levels of FeNO, exhaled carbon monoxide, total serum IgE, and higher eosinophilia. Boys had higher FeNO levels than girls. We found a significant positive correlation between FeNO levels and the percentage of blood eosinophils, %predicted of forced vital capacity, total serum IgE levels, and increasing age. CONCLUSIONS: Various phenotypes of children's asthma are characterized by specific pattern of the results of clinical and laboratory tests. FeNO correlates with total serum IgE, blood eosinophilia, age, and some spirometric parameters with different strength. Therefore, the coexistence of atopy, concomitant allergic rhinitis/rhinoconjunctivitis, and some other parameters should be considered in critical evaluation of FeNO in the management of asthmatic children.

Oxidative alternations in rat heart homogenate and mitochondria during ageing.

Our understanding of the role played by reactive oxygen and nitrogen species in disease pathology and ageing is still insufficient. Reactive oxygen species and reactive nitrogen species can initiate protein and lipid oxidative damage that may be the most important contribution to ageing and age-related heart diseases. In the present study, we investigated the effect of ageing on oxidative damage of protein amino acid residues and lipids in heart homogenate and mitochondria of 4- and 26-month-old Wistar rats. Levels of dityrosine and levels of lysine conjugates increased in heart homogenate during ageing, although levels of conjugated dienes did not change. We observed significantly oxidative modification of tryptophan in heart mitochondria and increased levels of dityrosine with advancing age. However, levels of lysine conjugates, conjugated dienes as well as relative level of cytochrome c oxidase were unchanged in heart mitochondria during ageing. The results of this study suggest a different mechanism of oxidative modification in heart compartments during ageing and moreover, mitochondria and other cellular compartments are targets for oxidative modifications.

Age-dependent effect of oxidative stress on cardiac sarcoplasmic reticulum vesicles.

The oxidative stress hypothesis of aging suggests that accumulation of oxidative damage is a key factor of the alterations in physiological function during aging. We studied age-related sensitivity to oxidative modifications of proteins and lipids of cardiac sarcoplasmic reticulum (SR) isolated from 6-, 15- and 26-month-old rats. Oxidative stress was generated in vitro by exposing SR vesicles to 0.1 mmol/l FeSO4/EDTA + 1 mmol/l H2O2 at 37 degrees C for 60 min. In all groups, oxidative stress was associated with decreased membrane surface hydrophobicity, as detected by 1-anilino-8-naphthalenesulfonate as a probe. Structural changes in SR membranes were accompanied by degradation of tryptophan and significant accumulation of protein dityrosines, protein conjugates with lipid peroxidation products, conjugated dienes and thiobarbituric acid reactive substances. The sensitivity to oxidative damage was most pronounced in SR of 26-month-old rat. Our results indicate that aging and oxidative stress are associated with accumulation of oxidatively damaged proteins and lipids and these changes could contribute to cardiovascular injury.

Exhaled carbon monoxide as a new marker of respiratory diseases in children.

Among modern methods included in diagnostic algorithms for various diseases, analyses of expired breath and its condensate acquire increasing importance. Various markers can be determined in the exhaled air, especially volatile gaseous compounds: nitrogen oxide (NO), carbon monoxide (CO), hydrocarbons and 8-isoprostanes. In contrast to NO, CO can serve as a marker of inflammation and oxidation stress. The representation of CO in the exhaled breath (eCO) changes in various diseases of the respiratory and other systems. Among the respiratory diseases in which the use of eCO measurement seems to be perspective and beneficial are bronchial asthma, airways infections, cystic fibrosis, and primary ciliary dyskinesia. The observation of eCO concentrations represents a modern, simple, available, and well reproducible method for the diagnosis of many diseases of respiratory system in children and for the observation of progression, severity of the disease, and response to therapy.

Food allergens and respiratory symptoms.

Food allergy may be clinically expressed by a variety of respiratory symptoms, which can be provoked either by IgE- or cellular mediated reactions. Among the diagnostic procedures, newly introduced atopy patch test seems to be important for diagnosis of cellular, delayed immune reactions. We studied the prevalence of positive atopy patch tests with food and inhalant allergens and the correlation between the positivity of atopy patch tests and questionnaire derived atopic and nonatopic espiratory symptoms and diseases in an unselected children population. We found a correlation between the positive patch test result with wheat and cough after physical effort, allergic rhino-conjunctivitis, and bronchitis recidivans. The subjects with positive skin reaction to egg suffered from allergic rhino-conjunctivitis and bronchial asthma. Food and inhalant allergens play an important role in the induction and exacerbation of some respiratory allergic diseases. The positive correlation of positive results of skin tests and history of some respiratory diseases and symptoms also on the population level confirm the importance of these tests in the diagnostic work-up of these allergic diseases.

Effect of aging on the expression of intracellular Ca(2+) transport proteins in a rat heart.

Aging process is accompanied by various biological dysfunctions including altered calcium homeostasis. Modified calcium handling might be responsible for changed cardiac function and potential development of the pathological state. In the present study we compared the mRNA and protein levels of the intracellular Ca(2+)-handling proteins--inositol 1,4,5-trisphosphate receptor (IP(3)R), ryanodine receptor (RyR), sarcoplasmic reticulum Ca(2+) pump (SERCA2), and also transient receptor potential C (TRPC) channels in cardiac tissues of 5-, 15-, and 26-month-old rats. Aging was accompanied by significant increase in the mRNA levels of IP(3)R and TRPC channels in both ventricles and atria, but mRNA level of the type 2 RyR was unchanged. Protein content of the IP(3)R1 correlated with mRNA levels, in the left ventricle of 15- and 26-month-old rats the value was approximately 1.8 and 2.8-times higher compared to 5-month-old rats. No significant differences were observed in mRNA and protein levels of the SERCA2 among 5-month-old and aged rats. However, Ca(2+)-ATPase activity significantly decreased with age, activities in 5-, 15-, and 26-month-old rats were 421.2 +/- 13.7, 335.5 +/- 18.1 and 304.6 +/- 14.8 nmol P(i) min(-1) mg(-1). These results suggest that altered transporting activity and/or gene expression of Ca(2+)-handling proteins of intracellular Ca(2+) stores might affect cardiac function during aging.

Age-related oxidative modifications of proteins and lipids in rat brain.

Oxidants have been shown to play a major role in ageing and ageing-related neurodegenerative diseases. In the present study, we investigated the effect of ageing on oxidative damage to lipids and proteins in brain homogenate, mitochondria and synaptosomes of adult (6-month-old), old (15-month-old), and senescent (26-month-old) Wistar rats. There was a significant increase in thiobarbituric acid-reactive substances and conjugated dienes in homogenates, which indicate increased lipid peroxidation (LPO). Oxidative modifications of homogenate proteins were demonstrated by a loss of sulfhydryl content, accumulation of dityrosines and formation of protein conjugates with LPO-end products. Increase in protein conjugates with LPO-end products and a decrease in SH groups were observed also in mitochondria and synaptosomes, but dityrosine content was elevated only in synaptosomes. Protein surface hydrophobicity, measured by fluorescent probe 1-anilino-8-naphthalenesulfonate (ANS), was increased only in homogenate. These results suggest that besides mitochondria and synaptosomes other cellular compartments are oxidatively modified during brain ageing.

Impact of Ginkgo Biloba Extract EGb 761 on ischemia/reperfusion - induced oxidative stress products formation in rat forebrain.

Dysbalance in reactive oxygen/nitrogen species is involved in the pathogenesis of cerebral ischemia/reperfusion injury (IRI). Ginkgo biloba extract (Egb 761) pre-treatment was used to observe potential antioxidant/neuroprotective effect after global ischemia/reperfusion. Egb 761 significantly decreased the level of lipoperoxidation (LPO) in rat forebrain total membrane fraction (homogenate) induced by in vitro oxidative stress (Fe(2+)+H(2)O(2)). In animals subjected to four-vessel global ischemia for 15 min and 2-24 h reperfusion the EGb pretreatment slightly decreased LPO in forebrain homogenate. However, as detected in EGb treated group, the LPO-induced lysine conjugates are attenuated in comparison to non-treated IRI animals. EGb significantly improved parameters which indicate forebrain protein oxidative damage after IRI. The intensity of tryptophane fluorescence was increased by the 18.2% comparing to non-treated IRI group and bityrosine fluorescence was significantly decreased in ischemic (21%) and 24 h reperfused (15.9%) group in comparison non-treated IRI group. In addition, the level of total free SH- groups in pre-treated animals was significantly higher comparing to non-treated animals. Our results indicate that extract of EGb 761 has potent antioxidant activity and could play a role to attenuate the IRI-induced oxidative protein modification and lipoperoxidation in the neuroprotective process.

Oxidative modification of rat cardiac mitochondrial membranes and myofibrils by hydroxyl radicals.

The effect of hydroxyl radicals generated by the FeSO4/H2O2 system on structural properties of proteins and membranes was studied in rat cardiac mitochondria and myofibrils. Exposure of mitochondria to 0.1 mmol/l FeSO4/EDTA plus 1 mmol/l H202 at 37 degrees C for 30 or 60 min caused conjugated diene formation, but it was not accompanied by accumulation of fluorescent lipid-protein conjugates. On the other hand, fluorescence measurements revealed radical-induced and time-dependent loss of tryptophans and production of bityrosines. Under the same conditions, the gradual decrease in tryptophan flurescence and increase in bityrosine formation was also observed in radical-treated myofibrils. These results suggest that *OH radicals can alter the mitochondrial and myofibrillar function via oxidation of amino acid residues and might be implicated in the pathogenesis of myocardial injury.

Molecular pathways of endoplasmic reticulum dysfunctions: possible cause of cell death in the nervous system.

This review summarizes recent information on the role of calcium in the process of neuronal injury with special attention to the role of calcium stores in the endoplasmic reticulum (ER). Experimental results present evidence that ER is the site of complex processes such as calcium storage, synthesis and folding of proteins and cell response to stress. ER function is impaired in many acute and chronic diseases of the brain which in turn induce calcium store depletion and conserved stress responses. Understanding the mechanisms leading to ER dysfunction may lead to recognition of neuronal protection strategies.