The Lack of Influence of Homozygous Long Allele of the 5-HTTLPR Gene on the Severity of Alcohol Craving During 6 Weeks of Rehab Hospitalisation in Comparison to Not Homozygous and Homozygous Short Alleles - Preliminary Report.

Purpose: The aim of this study was to assess changes in the severity of alcohol craving according to allelic variants of the 5-HTTLPR gene polymorphism during hospitalisation and their association with selected clinical variables in alcohol-dependent patients. Patients and Methods: The study is exploratory. Participants were investigated at the 2nd and 6th week of alcohol-dependence therapy in the addiction treatment unit. Recruitment was conducted among alcohol-dependent patients from several Polish drug treatment centres. The total sample size was 130 persons (12 females and 118 males). Study subjects' mean age was 43.0 years. Patients were investigated twice by using the Penn Alcohol Craving Scale (PACS) and Beck Depression Inventory (BDI), and once by using Short Alcohol Dependence Data Questionnaire (SADD) and taking a swab for genetic testing. The polymorphism of the gene encoding the serotonin transporter 5-HTTLPR (SLC6A4) was determined from isolated DNA and its homozygous variants of short/short or long/long alleles and heterozygous short/long alleles were analysed. Results: At 6th week of the follow-up, there was a decrease in the severity of alcohol craving in half of subjects with the short/short allele (p = 0.033) and in one-fifth of subjects with the long/short allele (p = 0.002) of the 5-HTTLPR gene. In subjects with long/long allele of the 5-HTTLPR gene, there was no change in the severity of alcohol craving between 2nd and 6th weeks of the study (p = 0.242). Conclusion: There was no statistical influence of the homozygous long allele of the 5-HTTLPR gene on severity of alcohol craving during 6 weeks of rehab hospitalisation in comparison to not homozygous and homozygous short alleles. The s-allele was associated with decrease of alcohol craving. It may point on the potential need for differentiated rehabilitation methods depending on the genetic diversity of addicted patients and its role in the severity of alcohol craving.

Confusion of Alcohol Craving With Food Hunger in Alcohol-Dependent Individuals Entering In-Patient Drug Treatment.

AIM: To identify the distinguishing characteristics of alcohol dependent patients who confuse alcohol cravings with pre-meal hunger. METHODS: Data were collected at interview on sociodemographic status, clinical status and anthropometry in 179 patients (163 men and 16 women) undergoing in-patient treatment for alcohol dependence. RESULTS: A comparison of the patient subgroups studied showed that patients who did not confuse, and those who did confuse, alcohol craving with pre-meal hunger differed significantly in terms of alcohol craving scale scores (9 vs. 4 points). Patients confusing alcohol cravings with pre-meal hunger were more likely to recognize that experiencing severe pre-meal hunger can cause relapse (67.9 vs. 22.8%) and that not being able to distinguish between the sensations under study also increases the risk of breaking abstinence (75.0% vs. 50.4%). This was independent of severity of dependence and intensity of recent alcohol consumption. CONCLUSIONS: Alcohol-dependent persons who confuse alcohol craving with pre-meal hunger differ from those who do not confuse these hunger pangs in terms of feeling stronger alcohol craving and more frequent occurrence of symptoms accompanying the feeling of alcohol craving during pre-meal hunger. At the start of treatment for alcohol withdrawal, alcohol-dependent individuals who report confusing alcohol cravings with pre-meal hunger are less confident of maintaining abstinence. This is relevant to treatment. The role of possible confounders (depressive symptoms, cognitive and educational deficiency) could not be elucidated definitively.

Position statement of the Polish Academy of Sciences Committee Human Nutrition Science on the principles for the nutrition of children aged 1-3 years.

The Position Statement on the principles of nutrition for children aged 1-3 years emphasizes that proper nutrition of children at this age determines their optimal psychometric development and has beneficial effects on the process nutritional programming, which reduces the risk of diet-related diseases in adulthood. Continued breastfeeding in the post-infancy period, together with the proper introduction of complementary foods, supplies all the nutritional needs of the child. A varied selection of food products is important to balance out the diet of a child in the context of energy and nutrient needs. Attention should be paid to products not recommended for frequent consumption, due to the possibility of the early development of improper eating habits that can lead to undesirable health consequences. Due to the potential risk of deficiency, adequate intake of iron, iodine, calcium and vitamin D, as well as of n-3 PUFAs (which is often insufficient) should be provided. Adequate dietary energy and protein intake protects children against protein-energy undernutrition and is crucial for their proper growth and development. An important element in the assessment of the development of children involves monitoring their nutritional status and physical development by systematically measuring their body weight and length/height and analyzing their weight gain. It is necessary to diagnose the causes of being underweight/overweight in children. Physical activity (such as outdoor walks, plays, and games) and healthy sleep hygiene are recommended. Physical activity, an adequate number of hours of sleep, and the quality of sleep in early childhood may improve immunity, reduce the risk of excessive weight gain, and consequently reduce the risk of obesity later in life. Other issues discussed include the functioning of the digestive system as one of the determinants of the nutrition of young children, basics of proper nutrition, risk of nutrient deficiencies and development of proper eating habits in early childhood.

Initial Study on COMT and DRD2 Gene Polymorphisms as Well as the Influence of Temperament and Character Trait on the Severity of Alcohol Craving in Alcohol-Dependent Patients.

The main aim of this work was to determine the impact of COMT and DRD2 gene polymorphisms together with temperament and character traits on alcohol craving severity alcohol-dependent persons. The sample comprised of 89 men and 16 women (aged 38±7). For the sake of psychological assessment various analytic methods have been applied like the Short Alcohol Dependence Data Questionnaire (SADD), Penn Alcohol Craving Scale (PACS) or Temperament and Character Inventory (TCI) test. The SNP polymorphism of the analyzed genes was determined by Real Time PCR test. The results showed, that the COMT polymorphismmay have an indirected relationship with the intensity and changes in alcohol craving during abstinence. The DRD2 receptor gene polymorphisms are related with the intensity of alcohol craving. It seems that the character traits like "self-targeting", including "self-acceptance", are more closely related to the severity of alcohol craving and polymorphic changes in the DRD2 receptor than temperamental traits. Although this is a pilot study the obtained results appeared to be promising and clearly indicate the link betweengene polymorphisms alcohol craving and its severity.

Enzymatic Activity and Physicochemical Properties of Soil Profiles of Luvisols.

Most studies on soil enzymes are focused on the upper horizons of the soil profile, even though they transform the soil organic matter at every depth of the soil profile. The aim of this work was to investigate the distribution of ß-glucosidase (GLU), nitrate reductase (NR), urease (UR), phosphatase (PHA), dehydrogenase (DHA) and catalase (CAT) activity through 14 trunked soil profiles of the Luvisols formed from a glacial till. The content of microbial biomass carbon (MBC) as well as physicochemical properties such as organic carbon (CORG), total nitrogen (NTOT), available P, K and Mg, soil density and porosity, pH in KCl and fractional composition were also studied. In general, enzymatic activity was highest in the top 30 cm layer of the profiles and decreased progressively towards the deeper horizons. The exceptions were the NR activity, which was active only in the Ap horizon and whose activity decreased sharply to nearly zero in the Bt horizon and parent rock, and the PHA activity, which was highly active even in the parent rock depth. The decreased availability of carbon and nutrients was the main driver of decreases in microbial abundance and enzymatic activity with depth. The enzymatic activity, when expressed on a CORG and MBC basis, behaves differently compared to the activity expressed on a soil mass basis. The activity decreased (NR), increased (PHA, UR), showed no clear pattern (GLU) or the changes were not significant (DHA, CAT). The content of CORG, NTOT, K and PAVAIL generally decreased with depth, while for Mg, there was no clear direction in the profile distribution. Future studies to characterize the substrate distribution within the soil profile and enzyme stability will provide further insight into the controls on nutrient cycling and related enzymes throughout the soil profiles.

The Molecular Basis of Alcohol Use Disorder (AUD). Genetics, Epigenetics, and Nutrition in AUD: An Amazing Triangle.

Alcohol use disorder (AUD) is a very common and complex disease, as alcohol is the most widely used addictive drug in the world. This disorder has an enormous impact on public health and social and private life, and it generates a huge number of social costs. Alcohol use stimulates hypothalamic-pituitary-adrenal (HPA) axis responses and is the cause of many physical and social problems (especially liver disease and cancer), accidental injury, and risky sexual behavior. For years, researchers have been trying to identify the genetic basis of alcohol use disorder, the molecular mechanisms responsible for its development, and an effective form of therapy. Genetic and environmental factors are known to contribute to the development of AUD, and the expression of genes is a complicated process that depends on epigenetic modulations. Dietary nutrients, such as vitamins, may serve as one these modulators, as they have a direct impact on epigenomes. In this review, we connect gathered knowledge from three emerging fields-genetics, epigenetics, and nutrition-to form an amazing triangle relating to alcohol use disorder.

Data-Driven Dietary Patterns and Diet Quality Scores: Reproducibility and Consistency in Sex and Age Subgroups of Poles Aged 15-65 Years.

This study aimed to assess: (i) the test-retest reproducibility of identification of data-driven dietary patterns (DPs) derived using a Principal Component Analysis (PCA) and hypothesis-driven DPs (diet quality scores); (ii) the consistency of data-driven DPs with diet quality scores in sex and age subgroups of Poles aged 15-65 years. The study involved 504 subjects (55.6% of females). Data on food consumption frequency (33 food items) were collected twice with a two-week interval using the Dietary Habits and Nutrition Beliefs Questionnaire (KomPAN®) in a self-administered version (test and retest). Two major data-driven DPs ('Prudent' and 'Western') were identified in the total sample, sex groups and four age groups separately from test and retest data. Two diet quality scores were analysed: Pro-Healthy-Diet-Index-10 (pHDI-10) and Non-Healthy-Diet-Index-14 (nHDI-14). Tucker's congruence coefficient indicated fair-to-good similarity of data-driven DPs between test and retest for all study subgroups, except for males. Across study subgroups, the intraclass correlation coefficient (ICC) between the test and retest ranged from 0.56 to 0.86 for 'Prudent' DP and 0.57 to 0.82 for 'Western' DP, with the lowest values in males. The ICC (test vs. retest) ranged from 0.84 to 0.88 for pHDI-10 and 0.75 to 0.88 for nHDI-14. Comparing the data-driven DPs and diet quality scores, the Spearman's correlations ranged from 0.63 to 0.93 between 'Prudent' DP and pHDI-10, and from 0.60 to 0.81 between 'Western' DP and nHDI-14. The test-retest reproducibility of data-driven DPs and diet quality scores and their consistency were acceptable in most of the study subgroups, with a tendency to be higher for pro-health than unhealthy DPs. Data-driven DPs were more reproducible in females than males. The reproducibility of diet quality scores tended to be better in males than females and was the highest in 25-44-year-olds. The KomPAN® questionnaire can be recommended to use data-driven DPs and diet quality scores to describe the habitual diet in people aged 15-65 years.

Anemarrhenae asphodeloides rhizoma Extract Enriched in Mangiferin Protects PC12 Cells against a Neurotoxic Agent-3-Nitropropionic Acid.

The rhizome of Anemarrhena asphodeloides Bunge, used in Traditional Chinese Medicine as a brain function-improving herb, is a promising source of neuroprotective substances. The aim of this study was to evaluate the protective action of xanthones from A. asphodeloides rhizomes on the PC12 cell line exposed to the neurotoxic agent-3-nitropropionic acid (3-NP). The xanthone-enriched fraction of the ethanolic extract of A. asphodeloides (abbreviated from now on as XF, for the Xanthone Fraction), rich in polyphenolic xanthone glycosides, in concentrations from 5 to 100 µg/mL, and 3-NP in concentrations from 2.5 to 15 mM, were examined. After 8, 16, 24, 48, and 72 h of exposure of cells to various combinations of 3-NP and XF, the MTT viability assay was performed and morphological changes were estimated by confocal fluorescence microscopy. The obtained results showed a significant increase in the number of cells surviving after treatment with XF with exposure to neurotoxic 3-NP and decreased morphological changes in PC12 cells in a dose and time dependent manner. The most effective protective action was observed when PC12 cells were pre-incubated with the XF. This effect may contribute to the traditional indications of this herb for neurological and cognitive complaints. However, a significant cytotoxicity observed at higher XF concentrations (over 10 µg/mL) and longer incubation time (48 h) requires caution in future research and thorough investigation into potential adverse effects.

Reproducibility of a Short-Form, Multicomponent Dietary Questionnaire to Assess Food Frequency Consumption, Nutrition Knowledge, and Lifestyle (SF-FFQ4PolishChildren) in Polish Children and Adolescents.

The aim of the study was to assess the reproducibility of a short-form, multicomponent dietary questionnaire (SF-FFQ4PolishChildren) in Polish children and adolescents. The study involved 437 children (6-10 years old) and 630 adolescents (11-15 years old) from rural and urban areas of Poland. The self-administered questionnaire was related to nutrition knowledge, dietary habits, active/sedentary lifestyle, self-reported weight and height, and socioeconomic data. The questionnaire was completed with a two-week interval-twice by parents for their children (test and retest for children), twice by adolescents themselves (adolescent's test and retest) and once by adolescents' parents (parent's test). The strength of agreement measured using the kappa statistic was interpreted as follows: 0-0.20 slight, 0.21-0.40 fair, 0.41-0.60 moderate, 0.61-0.80 good, and 0.81-1.00 excellent. Regarding the frequency of consumption of food items and meals, kappa statistics were 0.46-0.81 (the lowest: fruit/mixed fruit and vegetable juices; the highest: Energy drinks) in test-retest for children, 0.30-0.54 (fruit/mixed fruit and vegetable juices; breakfast, respectively) in adolescent's test-retest, 0.27-0.56 (the lowest: Sweets, fruit, dairy products; the highest: Breakfast) in adolescent's test and parent's test. Lower kappa statistics were found for more frequently consumed foods (juices, fruit, vegetables), higher kappa statistics were found for rarely consumed foods (energy drinks, fast food). Across study groups, kappa statistics for diet quality scores were 0.31-0.55 (pro-healthy diet index, pHDI) and 0.26-0.45 (non-healthy diet index, nHDI), for active/sedentary lifestyle items they were 0.31-0.72, for components of the Family Affluence Scale (FAS) they were 0.55-0.93, for BMI categories (based on self-reported weight and height) they were 0.64-0.67, for the nutrition knowledge (NK) of adolescents the kappa was 0.36, for the nutrition knowledge of children's parents it was 0.62. The Spearman's correlations for diet quality scores were 0.52-0.76 (pHDI) and 0.53-0.83 (nHDI), for screen time score they were 0.45-0.78, for physical activity score they were 0.51-0.77, for the FAS score they were 0.90-0.93, and for the NK score they were 0.68-0.80. The questionnaire can be recommended to evaluate dietary and lifestyle behaviors among children and adolescents.

Reproducibility of a Questionnaire for Dietary Habits, Lifestyle and Nutrition Knowledge Assessment (KomPAN) in Polish Adolescents and Adults.

The aim of the study was to evaluate the reproducibility of the Dietary Habits and Nutrition Beliefs Questionnaire (KomPAN) in Polish adolescents and adults, including the assessment of indexes developed based on the questionnaire. In total, the study involved 954 subjects aged 15⁻65 (53.9% females). Interviews using the interviewer-administered questionnaire (IA-Q) in healthy subjects (n 299) and the self-administered questionnaire (SA-Q) in healthy subjects (n 517) and outpatients (n 138) were conducted and repeated after two weeks. Considering the consumption frequency of 33 food items, the cross-classification (test-retest) agreement of classification into the same category obtained for IA-Q in healthy subjects ranged from 72.2% (fruit juices) to 91.6% (energy drinks); the kappa statistic was >0.60 for all food items. For SA-Q conducted in healthy subjects the cross-classification agreement ranged from 63.8% (vegetable oils, margarines, mixes of butter and margarines) to 84.7% (lard); the kappa statistic was >0.50 for all food items. For SA-Q in outpatients, the cross-classification agreement ranged from 42.0% (both fruit juices and white rice, white pasta, fine-ground groats) to 92.0% (energy drinks); the kappa statistic was ≥0.40 for 20/33 food items. The kappa statistic for lifestyle items ranged 0.42⁻0.96, and for the nutrition knowledge level it ranged 0.46⁻0.73. The questionnaire showed moderate to very good reproducibility and can be recommended to assess dietary habits, lifestyle and nutrition knowledge of healthy adolescents and adults and those suffering from chronic diseases, after validation and/or calibration study is carried out. The reproducibility of the interviewer-administered questionnaire was better than its self-administered version. The reproducibility of the self-administered questionnaire was better in healthy subjects than in outpatients.

Low molecular weight proteins and enzymes in the urine of patients with bladder cancer - a pilot study.

INTRODUCTION: A steady increase in cases of bladder cancer (BC) has been observed. Detection of unfavorable changes, especially in the early stages of disease, is crucial to medical procedure. There is still a need to search for new, non-invasive biomarkers of BC. The aim of this study was to estimate the levels of selected low molecular weight proteins (LMWP) and enzymes in the urine of patients at different BC stages and grades. MATERIAL AND METHODS: Urine samples from 46 patients with BC and 16 healthy controls were examined. We measured levels of LMWP such as: retinol-binding protein (RBP), ß2 -microglobulin (ß2M), enzymes: N-acetyl-ß-D-glucosaminidase (NAG), isoform (NAG-B) and also neutrophil gelatinase-associated lipocalin (NGAL). RESULTS: The levels of all examined parameters differed between patients and healthy subjects. Levels of NAG (p = 0.031), NAG-B (p = 0.023) and NGAL (p = 0.008), and total protein (p = 0.007) concentrations, were significantly higher in the BC patients than in the control group. Among the examined parameters, positive significant correlations were observed only between urinary NGAL concentration and tumor stages and grades. The highest percentages of changes in NGAL concentration were observed in tumor in situ (TIS) and G3grade patients. CONCLUSIONS: Our study showed that urinary NGAL concentrations, as well as NAG and NAG-B activity, could be helpful noninvasive parameters for the diagnosis of BC. The most promising seems to be NGAL determination, but further study is needed on a larger group of participants in order to confirm this observation.

Evaluation of NMP22 in bladder cancer patients sensitive to environmental toxins.

BACKGROUND: Bladder cancer (BC) is recognized as environmentally related. The interaction of environmental exposure to chemicals and genetic susceptibility seem to play important roles in BC development. In order to improve diagnosis and the recognition of BC risk, a group of markers which combine genetic susceptibility with detoxification and nuclear matrix protein (NMP22) is proposed. OBJECTIVES: The aim of the study was to examine the utility of nuclear matrix protein (NMP22) as a diagnostic marker in BC in genetic susceptibility (NAT2 slow acetylators) combined with detoxification abilities (glutathione S-transferase GST and isoenzyme GST-π). MATERIAL AND METHODS: The NMP22 level in urine, N-acetyltransferase 2 (NAT2) genotype and GST activity in hemolysate blood, as well as isoenzyme GST-π level, were determined in the urine and serum of 43 patients with BC and from 25 non-cancer controls. NMP22 and isoenzyme GST-π levels were measured by ELISA. The NAT2 genotype was examined in DNA isolated from whole blood using the PCR (Polymerase Chain Reaction) technique, while the activity of GST was determined with the spectrophotometric method. RESULTS: In the BC group, NMP22 (p = 0.005) concentration, GST-π (p = 0.003) in urine and GST (p = 0.009) activity in blood were statistically significantly higher than in the healthy controls. The majority of BC patients were slow acetylators (NAT2 genotype). A correlation between the level of nuclear matrix protein NMP22 and GST was found in all BC group (p = 0.007) and also slow acetylators (p = 0.0147). CONCLUSIONS: The results support the utility of a marker combination, which covers the genetic susceptibility to chemicals with the level of detoxification and nuclear matrix protein in BC patients. A relationship between NMP22 level in urine, GST level in blood and NAT2 genotype was observed. Also the isoenzyme GST-π in urine seems useful as a marker of BC.

The Diagnostic Value of Nuclear Matrix Proteins in Bladder Cancer in the Aspect of Environmental Risk from Carcinogens.

BACKGROUND: The interaction of environmental factors with genetic susceptibility and detoxification level seems to be an important causative factor in bladder cancer (BC). The aim of this study was to look for a BC marker panel which reflects the environmental risk. The nuclear matrix protein 22 (NMP22), bladder cancer-4 (BLCA-4), and total level proteins NMP22 and BLCA-4 (NMBL) in BC patients with genetic predisposition NAT2 (classified as slow acetylators, SA), DNA damage (8-OHdG), and detoxification by isoenzyme GSTπ activity were measured. MATERIALS AND METHODS: The urine and blood from 91 BC patients and controls were examined, also according to tumor stage (T) and grade (G). The participants completed a questionnaire in order to evaluate environmental risk. RESULTS: Most patients (75.3%) were previous or actual smokers. The levels of 8-OHdG, NMP22, BLCA-4, NMBL, and GSTπ were significantly higher in BC (p ≤ 0.001). The majority of patients (59.3%) were slow acetylators (SA). The highest BLCA-4/8-OHdG correlation was observed in total BC and SA smokers. CONCLUSIONS: The total pool of nuclear matrix proteins in the urine (NMBL) has a higher diagnostic value in bladder cancer than single proteins. The particular value of BLCA-4 and GSTπ in the aspect of environmental risk was noted.

The role of the BLCA-4 nuclear matrix protein in bladder cancer.

Bladder cancer (BC) affects usually older people. According to information provided by the National Cancer Registry in 2012. BC was the 4th, in terms of illness, cancer in men and 11th in women. Early diagnosis of bladder cancer is important because detected later has worse prognosis.Diagnosis of bladder cancer is not simple and it is still very invasive. Usually the cystoscopy or endoscopic bladder biopsy with histopathological evaluation and cytology of urine sediment is used. This prompted researchers to look for alternative noninvasive methods of diagnosis of bladder cancer. Recently, it was described the group of six proteins (BLCA) specific for BC, with special attention to BLCA-4.BLCA-4 belongs to the nuclear matrix protein and has a high specificity for this type of cancer however the value of this marker in BC diagnosis is not yet established. Oxidative DNA damage play an important role in the pathogenesis of some human diseases, including cancer. Determination of 8-hydroxy-2'deoksyguanozyne (8-OHdG) is currently used in the evaluation of genotoxic damage.The aim of the work was to review information on BLCA-4, its function in the process of BC carcinogenesis and diagnostic value also in exposure to genotoxic compounds measured by 8-hydroxy-2'deoksyguanozyne (8-OHdG) level.

The role of 17ß-estradiol metabolites in chromium-induced oxidative stress.

BACKGROUND: The increasing incidence of estrogen-dependent breast cancer and the presence in the environment of a large number of factors that interact with estrogen receptors have sparked interest in chemical influences on estrogen-dependent processes. In a previous work, the authors examined the interaction of estradiol with chromium. In the present article the importance of estradiol biotransformation in these interactions is investigated. There is no information in the available literature about the role of metabolites in exposure to chromium. It seems important because estradiol metabolites have various carcinogenic abilities and their formation during biotransformation could be increased or decreased by environmental enzyme inducers or inhibitors. The metabolites could play a detoxifying role or create a toxic synergism in free radical processes induced by chromium VI (CrVI). OBJECTIVES: The aim of this study was to evaluate the influence of 2 17ß-estradiol metabolites - 4-hydroxyestradiol (4-OHE2) and 16α-hydroxyestrone (16α-OHE1) - in conditions of oxidative stress caused by CrVI. MATERIAL AND METHODS: Human blood, erythrocytes or mitochondria isolated from human placentas after natural deliveries were used in the experiments. The influence of CrVI, 4-OHE2 and 16-OHE1 on thiobarbituric acid reactive substances (TBARS), the hydroxyl radical (•OH), superoxide dismutase (SOD), glutathione peroxidase (GPx) and glutathione-S-transferase (GST), and the interactions of the metabolites exposed to chromium expressed by these factors were examined. RESULTS: 4-OHE2 reduced the level of TBARS induced by CrVI in mitochondria (p < 0.05) and in erythrocytes (p < 0.05), and increased SOD activity (p < 0.05). 16α-OHE1 increased the activity of GST in erythrocytes exposed to CrVI (p < 0.05). CONCLUSIONS: The metabolites do not have toxic interactions with CrVI. On the contrary, they exhibited a protective effect. The mechanism of protection varied: 4-OHE2 decreased TBARS and increased SOD activity, while 16α-OHE1 increased GST activity.

Glutathione S - transferases class Pi and Mi and their significance in oncology.

In this article the current data, which shows that glutathione S-transferases (GST) class Pi and Mi are interesting and promising biomarkers in acute and chronic inflammatory processes as well as in the oncology, were presented based on the review of the latest experimental and clinical studies. The article shows their characteristics, functions and participation (direct - GST Pi, indirect - GST Mi) in the regulation of signaling pathways of JNK kinases, which are involved in cell differentiation. Overexpression of glutathione S-transferases class Pi and Mi in many cancer cells plays a key role in cancer treatment, making them resistant to chemotherapy. GST isoenzymes are involved in the metabolism of various types of xenobiotics and endogenous substrates, so their altered expression in cancer tissues as well as in serum and urine could be an important potential marker of the cancer and an indicator of oxidative stress. The study shows the role of glutathione S-transferases in redox homeostasis of tumor cells and in the mechanism of resistance to anticancer drugs.

THE ESTROGENS / CHROMIUM INTERACTION IN THE NITRIC OXIDE GENERATION.

The interaction of estrogens with environmental toxins in free radicals generation: reactive oxygen species (ROS) or reactive nitrogen species (RNS) which participates in cancerogenesis is not yet recognized. Chromium(VI) is widely present in environment. One of its toxicity pathway is free radicals generation. Estrogens have the ability to scavenge free radicals, but may also act as prooxidants. Both chromium(VI) and estrogens are classified by International Agency for Research on Cancer (IARC) as carcinogens, so synergistic effect seems very dangerous. The interaction of chromium and estrogens in ROS generation are partly described but there are no reports on estrogen/chromium interaction on nitric oxide (NO) generation. The aim of the study was to examine the interaction of chromium(VI) and 17-p-estradiol (E2) on NO level in human blood as well as the role of E2 metabolites: 4-hydroxyestradiol (4-OHE2) and 16a-hydroxyestrone (16α-OHE1) in these processes. The NO level was estimated with the diagnostic kit (Nitric Oxide Colorimetric Detection Kit from Arbor Assays) in human blood in vitm. The results showed that Cr(VI) in used concentration (0.5; 1.0 and 5.0 gg/mL) decreases significantly NO level in blood, acting antagonistically to E2 and 4-OHE2. Estrogens (E2, 4-OHE2 and 16α-OHEI) do not protect against inhibiting effect of Cr(VI) on nitric oxide generation in blood because after combined exposure the decreased production of NO in blood was noted. In conclusion, presented results provide the information about the character of estrogen/Cr(VI) interaction in NO level in human blood. It is important knowledge for cardio protected effect e.g., hormone replacement therapy in environmental or occupational exposure to Cr(VI), chromium supplementation, also important for cancer risk evaluation.

SYNTHESIS AND ANTIBACTERIAL ACTIVITY OF NEW SULFONAMIDE ISOXAZOLO[5,4-b]PYRIDINE DERIVATIVES.

A series of novel sulfonamide isoxazolo[5,4-b]pyridines were synthesized. The substrates for their synthesis were 3-aminoisoxazolo[5,4-b]pyridine and selected aryl sulfonic chlorides, chlorosulfonic acid and selected amines. Reactions were carried out using the classical and microwave methods. Selected compounds were tested towards antibacterial and antiproliferative activity. The structure of the obtained new derivatives was determined by elemental analysis and acquired IR and 1H NMR spectra. Among the tested compounds: N- isoxazolo[5,4-b]pyridine-3-yl-benzenesulfonamide (2) and N-isoxazolo[5,4-b]pyridine-3-yl-4-methylbenzene-sulfonamide (5) showed antimicrobial activity towards Pseudomonas aeruginosa (ATCC 27853) and Escherichia coli (ATCC 25922) at doses: 125, 250 and 500 µg. Both compounds showed a 50% inhibition of proliferation of breast carcinoma cell line MCF7 at concentrations of 152.56 µg/mL and 160 161.08 µg/mL, respectively.

Preliminary Study on J-Resolved NMR Method Usability for Toxic Kidney's Injury Assessment.

BACKGROUND: Nowadays, the Nuclear Magnetic Resonance (NMR) techniques are tested for metabolomic urine profile in order to detect early damage of kidney. OBJECTIVES: The purpose of this investigation was the initial assessment of two-dimensional J-resolved NMR urine spectra analysis usability for early kidney injuries detection. The amino acids (AA) and acids profile change after the exposure to nephrotoxic agent (the cisplatin infusion) was examined. MATERIAL AND METHODS: The material was the urine of patients with non-small-cell lung cancer, treated with cisplatin in Pulmonology and Lung Cancers Clinic in Wroclaw. The urine of healthy volunteers was also examined. The identification of metabolites in urine was based on two-dimensional JRES signals in spectra, described in Human Metabolites Database (HMD). The molar concentration of metabolites was calculated from the volume under the signals. The analysis was focused on amino acids and organic acids (lactid acid and pyruvic acid) profiles. RESULTS: Any specific amino acids were identified after cisplatin infusion in comparison to the state before infusion. However, the differences in concentration were observed over 2-fold increase in valine, isoleucine and leucine, over 3-fold in alanine. Also, the concentration of pyruvic and lactic acids increased significantly (p≤0.05, p≤0.01). CONCLUSIONS: There were no specific amino acids identified in response to the infusion of cisplatin; however, some changes in the concentrations of amino acids and other small molecules were found. The analysis of two-dimensional JRES spectra showed an increase of alanine, leucine, isoleucine and valine concentration after the application of cisplatin. It seems that it is worth developing the JRES method based on special computer program.