Organic amendments improve salinity-induced osmotic and oxidative stress tolerance in Okra (Abelmoschus esculentus (L.)Moench).

AIMS: Salinity adversely affects okra [Abelmoschus esculentus (L.) Moench] plants by inducing osmotic and oxidative stresses. This study was designed to enhance salinity-induced osmotic and oxidative stress tolerance in okra plants by applying organic amendments. METHODS: The effects of different organic amendments (municipal solid waste compost, farmyard manure (FYM) and press mud) on osmotic potential, water use efficiency, activities of antioxidant enzymes, total soluble sugar, total soluble proline, total soluble protein and malondialdehyde (MDA) contents of okra plants grown under saline conditions (50 mM sodium chloride) were evaluated in a pot experiment. The organic amendments were applied each at the rate of 5% and 10% per pot or in various combinations (compost + FYM, FYM + press mud and compost + press mud each at the rate of 2.5% and 5% per pot). RESULTS: As compared to control, high total soluble sugar (60.41), total soluble proline (33.88%) and MDA (51%) contents and increased activities of antioxidant enzymes [superoxide dismutase (83.54%), catalase (78.61%), peroxidase (53.57%] in salinity-stressed okra plants, were indicative of oxidative stress. Salinity significantly reduced the osmotic potential (41.78%) and water use efficiency (4.75%) of okra plants compared to control. Under saline conditions, 5% (farmyard manure + press mud) was the most effective treatment, which significantly improved osmotic potential (27.05%), total soluble sugar (4.20%), total soluble protein (73.62%) and total soluble proline (23.20%) contents and superoxide dismutase activity (32.41%), compared to saline soil. Application of 2.5% (FYM + press mud), 5% press mud, and 10% compost significantly reduced MDA content (27%) and improved activities of catalase (38.64%) and peroxidase (48.29%), respectively, compared to saline soil, thus facilitated to alleviate oxidative stress in okra plants. CONCLUSIONS: Using organic amendments (municipal solid waste compost, farmyard manure and press mud) was a cost-effective approach to improve salinity-induced osmotic and oxidative stress tolerance in okra plants.

Vibrational Corrections to NMR Spin-Spin Coupling Constants from Relativistic Four-Component DFT Calculations.

Zero-point vibrational (ZPV) corrections to the nuclear spin-spin coupling constants have been calculated using four-component Dirac-Kohn-Sham DFT for H2X (where X = O, S, Se, Te, Po), XH3 (where X = N, P, As, Sb, Bi), and XH4 (where X = C, Si, Ge, Sn, and Pb) molecules and for HC≡CPbH3. The main goal was to study the influence of relativistic effects on the ZPV corrections and thus results calculated at relativistic and nonrelativistic approaches have been compared. The effects of relativity become notable for the ZPV corrections to the spin-spin coupling constants for compounds with lighter elements (selenium and germanium) than for the spin-spin coupling constants themselves. In the case of molecules containing heavier atoms, for instance BiH3 and PbH4, relativistic effects play a crucial role on the results and approximating ZPV corrections by the nonrelativistic results may lead to larger errors than omitting ZPV corrections altogether.

Relativistic Four-Component DFT Calculations of Vibrational Frequencies.

We investigate the effect of relativity on harmonic vibrational frequencies. Density functional theory (DFT) calculations using the four-component Dirac-Coulomb Hamiltonian have been performed for 15 hydrides (H2X, X = O, S, Se, Te, Po; XH3, X = N, P, As, Sb, Bi; and XH4, X = C, Si, Ge, Sn, Pb) as well as for HC≡CPbH3. The vibrational frequencies have been calculated using finite differences of the molecular energy with respect to geometrical distortions of the nuclei. The influences of the choice of basis set, exchange-correlation functional, and step length for the numerical differentiation on the calculated harmonic vibrational frequencies have been tested, and the method has been found to be numerically robust. Relativistic effects are noticeable for the heavier congeners H2Te and H2Po, SbH3 and BiH3, and SnH4 and PbH4 and are much more pronounced for the vibrational modes with higher frequencies. Spin-orbit effects constitute a very small fraction of the total relativistic effects, except for H2Te and H2Po. For HC≡CPbH3 we find that only the frequencies of the modes with large contributions from Pb displacements are significantly affected by relativity.

The role of the electrokinetic charge of neurotrophis-based nanocarriers: protein distribution, toxicity, and oxidative stress in in vitro setting.

BACKGROUND: The rational chemical design of nanoparticles can be readily controlled and optimized by quantitatively studying protein adsorption at variously charged polymer carriers, determining their fate in biological fluids. We manufactured brain-derived neurotrophic factor (BDNF) -based electrostatic nanocomplexes with a different type of dendrimer core (anionic or cationic), encapsulated or not in polyethylene glycol (PEG), and studied their physicochemical properties and behavior in a biological setting. We investigated whether the electrokinetic charge of dendrimer core influences BDNF loading and desorption from the nanoparticle and serves as a determinant of nanoparticles' behavior in in vitro setting, influencing mitochondrial dysfunction, lipid peroxidation, and general nanoparticles' cellular toxicity. RESULTS: We found that the electrokinetic charge of the dendrimer core influences nanoparticles in terms of BDNF release profile from their surfaces and their effect on cell viability, mitochondrial membrane potential, cell phenotype, and induction of oxidative stress. The electrostatic interaction of positively charged core of nanoparticles with cell membranes increases their cytotoxicity, as well as serious phenotype alterations compared to negatively charged nanoparticles core in neuron-like differentiated human neuroblastoma cells. Moreover, PEG adsorption at nanoparticles with negatively charged core presents a distinct decrease in metabolic cell activity. On the contrary, charge neutralization due to PEG adsorption on the surface of nanoparticles with positively charged core does not reduce their cytotoxicity, makes them less biocompatible with differentiated cells, and presumably shows non-specific toxicity. CONCLUSIONS: The surface charge transformation after adsorption of protein or polyelectrolyte during nanocarriers formulation has an important role not only in designing nanomaterials with potent neuroprotective and neuroregenerative properties but also in applying them in a cellular environment.

NMR Spin-Spin Coupling Constants Derived from Relativistic Four-Component DFT Theory-Analysis and Visualization.

An unambiguous assignment of coupling pathways plays an important role in the description and rationalization of NMR indirect spin-spin coupling constants (SSCCs). Unfortunately, the SSCC analysis and visualization tools currently available to quantum chemists are restricted to nonrelativistic theory. Here, we present the theoretical foundation for novel relativistic SSCC visualization techniques based on analysis of the SSCC densities and the first-order current densities induced by the nuclear magnetic dipole moments. Details of the implementation of these techniques in the ReSpect program package are discussed. Numerical assessments are performed on through-space SSCCs, and we choose as our examples the heavy-atom Se-Se, Se-Te, and Te-Te coupling constants in three similar molecules for which experimental data are available. SSCCs were calculated at the nonrelativistic, scalar relativistic, and four-component relativistic density functional levels of theory. Furthermore, with the aid of different visualization methods, we discuss the interpretation of the relativistic effects, which are sizable for Se-Se, very significant for Se-Te, and cannot be neglected for Te-Te couplings. A substantial improvement of the theoretical SSCC values is obtained by also considering the molecular properties of a second conformation.

The intensification of anticancer activity of LFM-A13 by erythropoietin as a possible option for inhibition of breast cancer.

Recombinant human erythropoietin (Epo) is an effective and convenient treatment for cancer-related anaemia. In our study for the first time, we evaluated the effect of simultaneous use of Epo and Bruton's tyrosine kinase (BTK) inhibitor LFM-A13 on the viability and tumour development of breast cancer cells. The results demonstrated that Epo significantly intensifies the anticancer activity of LFM-A13 in MCF-7 and MDA-MB-231. The featured therapeutic scheme efficiently blocked the tumour development in zebrafish experimental cancer model. Epo and LFM-A13 administered together resulted in effective cell killing, accompanied by attenuation of the BTK signalling pathways, loss of mitochondrial membrane potential (MMP), accumulation of apoptotic breast cancer cells with externalised PS, a slight increase in phase G0/G1 and a reduction in cyclin D1 expression. Simultaneous use of Epo with LFM-A13 inhibited early stages of tumour progression. This therapeutic scheme may be rationale for further possible research.

Analysis of Base Monomer Elution from 3 Flowable Bulk-Fill Composite Resins Using High Performance Liquid Chromatography (HPLC).

BACKGROUND The aim of this study was to evaluate the elution of BisGMA, UDMA, TEGDMA, and HEMA monomers from flowable bulk fill composite resins with different resin matrix compositions, polymerized in 4-mm-thick layers, into 3 elution media. MATERIAL AND METHODS Three bulk-fill (SDR® (SDR), X-tra base (XB) and BEAUTIFIL-Bulk Flowable (BF)) resin-composites were tested. Cylindrical samples were immersed in 100% ethanol, 75% ethanol, and distilled water. The concentrations of the monomers were measured using the HLPC method (Agilent Technologies 1200 Series) after 1 and 24 h, as well as after 3, 7, 14, and 21 days. RESULTS After polymerization of the tested resins, there was elution of the BisGMA, UDMA, TEGDMA, and HEMA monomers from the SDR and BF composites, but none of the tested monomers could be detected eluting from XB. The highest penetrations of the polymerized SDR and BF composites were observed in the 100% ethanol solution. This extraction medium eluted the highest amounts of free monomers. Some eluted monomers were not described in the composites Material Safety Data Sheets. CONCLUSIONS The elution of the residual monomers depended on the resin composition and the materials filler/resin matrix ratio. In composite materials, toxicity assessment should be carried out, and should consider both the material composition as given by the manufacturer, and also the residual monomers that elute from the polymerized material. The elution concentration and time of monomers from composites depended on the solvent used. The highest penetrations of the polymerized SDR and BF composites were observed in the 100% ethanol solution, and this extraction medium eluted the highest amounts of free monomers. The 75% ethanol was a more aggressive medium than water in terms of monomer elution from bulk fill composites.

Inflammation markers are associated with metabolic syndrome and ventricular arrhythmia in patients with coronary artery disease.

BACKGROUND: Inflammation plays a major role in the development and progression of atherosclerosis and coronary artery disease (CAD). Inflammation markers, including white blood cell (WBC) count, C-reactive protein (CRP) and interleukin-6 (IL-6), are widely used for cardiovascular risk prediction. The aim of the study was to establish factors associated with WBC, CRP and IL-6 in patients with CAD. Two functional polymorphisms in genes encoding enzymes participating in adenosine metabolism were analyzed (C34T AMPD1, G22A ADA). METHODS: Plasma concentrations of IL-6 were measured using high-sensitivity ELISA kits, and the nephelometric method was used for high-sensitivity CRP (hs-CRP) measurement in 167 CAD patients. RESULTS: Presence of metabolic syndrome (MS) and its components, presence of heart failure, severity of CAD symptoms, severe past ventricular arrhythmia (sustained ventricular tachycardia [sVT] or ventricular fibrillation [VF]), lower left ventricle ejection fraction, higher left ventricle mass index, higher end-diastolic volume and higher number of smoking pack-years were significantly associated with higher WBC, CRP and IL-6. Strong associations with arrhythmia were observed for IL-6 (median 3.90 vs 1.89 pg/mL, p<0.00001) and CRP concentration (6.32 vs 1.47 mg/L, p=0.00009), while MS was associated most strongly with IL-6. CRP and IL-6 were independent markers discriminating patients with sVT or VF. There were no associations between AMPD1 or ADA genotypes and inflammation markers. CONCLUSIONS: WBC, CRP and IL-6 are strongly associated with components of the metabolic syndrome. Their strong association with life-threatening ventricular arrhythmia emphasizes the proarrhythmic role of inflammation in the increased cardiovascular risk of CAD patients.

Is plasma soluble CD36 associated with cardiovascular risk factors in early onset coronary artery disease patients?

BACKGROUND AND PURPOSE: This is the first study to investigate the relationship between plasma concentration of soluble CD36 (sCD36) and CD36 gene polymorphisms as well as clinical and echocardiographic parameters in patients with early onset coronary artery disease (CAD). METHODS: sCD36 concentrations were measured by the ELISA kits. CD36 sequence alterations detected by the DHPLC technique comprised single nucleotide substitutions: rs3173798, rs3211892, rs5956 and rs141680676. RESULTS: There were significant negative correlations between sCD36 and red blood cell count, hemoglobin, hematocrit and glucose concentration, ApoB/ApoA1 ratio, patients' weight and waist circumference, BMI, WHR, systolic blood pressure, MAP values, left ventricular end-diastolic diameter and volume, left atrium diameter, right ventricular end-diastolic diameter. There were significant positive correlations between sCD36 and patients' age, mean corpuscular volume of erythrocytes, HDL-cholesterol, ApoA1 concentrations. Significantly higher CD36 plasma levels were found in female subgroup. There was no association between CD36 genotypes and sCD36 concentrations. Multiple linear regression analysis revealed that significant independent predictors of higher plasma sCD36 level were female gender, older age, lower serum glucose and lower RBC. CONCLUSION: The presented data suggest possible protective effects of higher sCD36 concentration in relation to metabolic syndrome components in CAD patients. Higher sCD36 concentration is also associated with lower risk of left ventricular hypertrophy, but on the other hand is a potential risk factor of impaired left ventricle diastolic function.

Influence of daily diet on ascorbic acid supply to students.

BACKGROUND: Researchers suspect that the accepted adequate ascorbic acid plasma concentration is not being met even after dietary intake of the recommended amount of vitamin C. Current dietary intake recommendation in Poland is 60 mg per day for women and 75 mg per day for man (EAR), while in Western Europe and North America is higher and amounts to 75-90 mg per day. OBJECTIVE: The paper aimed at studying a correlation between composition of nutrients in daily diet and plasma vitamin C levels in university students. Materials and methods. This study examined diet composition and the nutritional status of ascorbic acid in plasma of 120 university students in Szczecin, Poland. Ascorbic acid was determined in blood plasma using HPLC method. The information concerning diet composition was collected using the method of "7-days food record" prior to blood collection. RESULTS: Plasma ascorbic acid deficiency (<40 µmol/L) was observed in 23% of women and 28% of men. The average plasma ascorbic acid concentration was 48.65 µmol/L in women and 45.61 µmol/L in men. The average intake of vitamin C in women with observed deficiency was average 46.55 mg/day, whereas in men it was 48.56 mg/day. CONCLUSIONS: The recommendation of dietary intake of vitamin C in Poland is low in comparison to other countries. Population- based studies are necessary to determine the actual demand for vitamin C in various population groups in Poland. Key words: nutrition, vitamins, dietary intake, diet, ascorbic acid, plasma.

Polymorphism of the CD36 Gene and Cardiovascular Risk Factors in Patients with Coronary Artery Disease Manifested at a Young Age.

This study investigates potential associations between CD36 gene variants and the presence of risk factors in Caucasians with coronary artery disease (CAD) manifested at a young age. The study group consisted of 90 patients; the men were ≤ 50 years old and the women were ≤ 55 years old. Amplicons of exons 4 and 5 including fragments of introns were analyzed by DHPLC. Two polymorphisms were found: IVS3-6 T/C (rs3173798) and IVS4-10 G/A (rs3211892). The C allele of the IVS3-6 T/C polymorphism was associated with higher prevalence of obesity and diabetes, higher hsCRP, lower Lp(a) serum concentrations, and younger age at myocardial infarction. The A allele of the IVS4-10 G/A polymorphism was associated with older age of myocardial infarction and higher white blood cell count. The functional role of CD36 polymorphisms in CAD development needs further research.

Oxidative stress modulates the organization of erythrocyte membrane cytoskeleton.

BACKGROUND: Apart from their main role in transporting oxygen and carbon dioxide, erythrocytes play also an important role in organism antioxidative defence. Direct exposure to reactive oxygen species (ROS) results in shortening of their half-life, even by 50%. The presence of glucose, being the substrate in pentose phosphate pathway (PPP) cycle, is one of the factors that can have influence on the level of oxidative stress. The activity of PPP increases during oxidative stress. Glucose guarantees normal PPP functioning with the production of reductive equivalents in the amounts necessary to reproduction of glutathione--nonenzymatic free radical scavenger. In available literature there are no reports regarding the changes in protein contents of erythrocyte cytoskeleton exposed to t-butyl hydroperoxide in relation to glucose presence in incubation medium. MATERIAL/METHODS: Erythrocytes taken from 10 healthy subjects were used to assess the influence of generated free radicals on erythrocyte proteins and chosen parameters of oxidative stress. Erythrocytes were incubated in the solutions containing deferent concentrations of t-butyl hydroperoxide and glucose. Electrophoresis was performed on polyacrylamide gel in denaturating conditions. The contents of tryptophan in membranes was evaluated spectrofluorometrically. RESULTS/CONCLUSIONS: In vitro conditions oxidative stress leads to protein damage in erythrocyte cytoskeleton, both in proteins inside the cell as well as having contact with extracellular environment. In consequence, the amount of low-molecular proteins--mainly globin, which bind to cytoskeleton, increases. This process takes place independently of glucose presence in incubation medium. One of the element of protein cytoskeleton, tryptophan, also undergoes degradation. The decrease of its contents is higher during erythrocyte exposure to t-BOOH in environment containing glucose, what can suggest prooxidative influence of glucose in conditions in vitro.

Neutrophil infiltration combined with necrosis in the primary tumor is a useful prognostic indicator for three-year disease-free survival time in patients with colorectal cancer.

Histopathological evaluation plays a key role in the diagnosis of colorectal cancer (CRC). Tumor-related local inflammation is regarded as a novel prognostic parameter. Neutrophils constitute one of the main types of inflammatory cells. The aim of the present study was to evaluate the prognostic value of intratumoral tumor-associated neutrophils (intraTANs), stromal TANs (stromaTANs) and necrosis, as well as their combined parametric value in formalin-fixed paraffin-embedded tissue sections from patients with CRC. For this purpose, a retrospective study of 160 patients with CRC who underwent surgery was conducted. The association of intraTANs, stromaTANs, necrosis and their combined parametric value with the clinicopathological features of patients with CRC was examined. The Kaplan-Meier method and the log-rank test were used to compare survival curves. To identify independent prognostic factors, uni- and multivariate Cox proportional hazards regression models were used. StromaTANs were associated with lymph node metastasis (P=0.049) and tumor deposits (P=0.041). In addition, necrosis was found to be associated with venous (P=0.003), lymphatic (P=0.007) and perineural (P=0.015) invasion, as well as with lymph node metastasis (P=0.033), the number of invaded lymph nodes (P=0.012), and lymph node pouch invasion (P=0.043). Furthermore, necrosis was found to be associated with the white blood cell count (P=0.030), neutrophil count (P=0.011), the combined neutrophil-to-lymphocyte ratio (NLR) and platelet-to-lymphocyte ratio (NLR-PLR) (P=0.038), and the combined platelet and NLR (PLT-NLR status) (P=0.030), as well as with the serum carcinoembryonic antigen (CEA) levels following surgery (P=0.011) and the monocyte-to-lymphocyte ratio (P=0.023). The combined parametric value was found to be associated with pT stage (P=0.049), venous (P=0.034) and lymphatic (P=0.026) invasion, and with serum CEA levels prior to surgery (P=0.029). The analysis of the 3-year disease-free survival (DFS) time revealed that tumor growth [hazard ratio (HR), 2.070; 95% CI, 1.837-3.808; P=0.003] and the combined parametric value (intraTANs, stromaTANs and necrosis, HR, 1.577; 95% CI, 1.372-3.032; P=0.028) were independent factors for patients with CRC. Taken together, the findings of the present study demonstrated that the combined value of neutrophils and necrosis examined in the cancerous tissue may be used as a prognostic factor for the 3-year DFS time in patients with CRC.

The Role of Oxidative Stress in the Risk of Cardiovascular Disease and Identification of Risk Factors Using AIP and Castelli Atherogenicity Indicators in Patients with PCOS.

Polycystic ovarian syndrome (PCOS) is one of the most common endocrinopathies in females of reproductive age and may affect 5-14% of women. In women with PCO syndrome, metabolic disorders such as insulin resistance, hyperinsulinemia, obesity, diabetes mellitus, and other elements of metabolic syndrome may occur. Patients with PCOS often have overweight and obesity, especially abdominal obesity, which is one of the risk factors for developing atherosclerosis. The atherogenicity indicators of AIP (atherogenic index of plasma) and Castelli's index are used to assess the risk of developing atherosclerosis. Studies have shown an increase in the concentration and activity of oxidative stress markers in patients with PCOS compared to women without the disease. The aim of the present study was to evaluate oxidative stress parameters in patients with PCOS in relation to insulin resistance, BMI, and hyperandrogenemia and to correlate them with cardiovascular risk parameters. Conclusions: The severity of oxidative stress in women with PCOS correlates with exposure to cardiovascular diseases. The assessment of additional cardiovascular disease (CVD) parameters is useful in identifying the risk groups for cardiometabolic disease among PCOS patients. When additional risk factors such as hyperandrogenism and insulin resistance (IR) are present in patients with PCOS, it is reasonable to include preventive examinations early. It is also important to evaluate lipidograms, which will make it possible to determine indicators of atherogenicity. Patients with PCOS and IR are at particular risk for cardiovascular complications. PCOS should be considered an important risk factor for CVD, which occurs independently of the occurrence of obesity. This factor is related to the important role of insulin resistance, which occurs independently of obesity. Atherogenic factors (AIP and Castelli index) are useful additional parameters to assess the risk of cardiometabolic disease in PCOS patients, especially among groups with insulin resistance. The early detection of risk factors should be an integral part of the care of PCOS patients. In laboratory studies of women with PCOS, TG, TChol, HDL-c and LDL-c levels, and glutathione peroxidase (GPx) activity were most clearly correlated with exposure to cardiovascular disease.

Assessment of the Parameters of Oxidative Stress Depending on the Metabolic and Anthropometric Status Indicators in Women with PCOS.

Polycystic ovary syndrome (PCOS) is one of the most common endocrinopathies in females of reproductive age. In women with PCOS, metabolic disorders such as insulin resistance (IR), hyperinsulinemia, obesity, diabetes mellitus, and other elements of metabolic syndrome are likely to occur. Studies have shown an increase in the concentration and activity of oxidative stress (OS) markers in patients with PCOS, compared to that in unaffected women. The aim of this study was to evaluate the parameters of OS in PCOS and their activity in relation to women without menstrual disorders with a normal body weight. Then, we compared malonodialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx), including overweight and obesity, hyperandrogenemia, and IR in the PCOS group. The study included 35 women aged 18-46, hospitalized for menstrual disorders in the form of infrequent menstruation. In 26 women, PCOS was diagnosed on the basis of the Rotterdam Criteria; these patients qualified for the study group. The control group (n = 21) consisted of patients without menstrual disorders and without PCOS in an ultrasound examination. Patients were diagnosed between the 2nd and 5th day of the cycle. The parameters of OS were analyzed and compared with the anthropometric parameters and the lipid profile of the patients. Enzymatic activity of GPx, CAT, SOD, and MDA levels was determined in both groups. MDA levels and CAT activity differed significantly between the groups. There was a decrease in MDA levels in the IR group and the involvement of GPx in the excess weight and obesity and IR group accompanied by an increase in hip circumference. It therefore seems that IR may be the main risk factor to exposure to OS in patients with PCOS, independent from obesity. In addition, GPx is involved in every step in the development of the pathological condition in PCOS.

The effect of L-ascorbic acid and/or tocopherol supplementation on electrophysiological parameters of the colon of rats chronically exposed to lead.

BACKGROUND: The aim of this study was to assess the effect of diet supplementation with L-ascorbic acid (500 mg/L), tocopherol (3 mg/kg b.w.), and/or a water soluble analog of tocopherol (Trolox) (48 mg/L) on ion transport in the colon of rats subjected to a chronic exposure (9 months) to 0.1% lead acetate in drinking water. MATERIAL/METHODS: The electrophysiological parameters of the colon wall were measured with Ussing methods. Lead content in the whole blood was analyzed by graphite furnace atomic absorption spectrometry (GFAAS) using Zeeman correction. L-ascorbic acid and tocopherol in plasma was measured by high performance liquid chromatography. Immunohistochemical reaction was carried out for visualization of occludin, the intracellular tight junction protein. RESULTS: We showed a strong inhibitory effect of lead on the electrophysiological parameters, changes in intestinal permeability, disappearance of junctional occludin, decreased amount of mucus covering the colon surface, and the accumulation of PAS-positive substance in the apical region of the cytoplasm in the absorptive cells. CONCLUSIONS: Supplementation with tocopherol or Trolox did not exert a beneficial influence on the studied parameters. L-ascorbic acid positively influenced the examined electrophysiological parameters, as it cancelled the inhibitory influence of lead on ion transport in the rat colon. L-ascorbic acid also protected against tight junction disruption of epithelial cells in the colon of the lead-treated rats. A similar effect was observed in the group of rats receiving lead and supplemented with L-ascorbic acid plus Trolox.

Hymenolepis diminuta: Activity of anti-oxidant enzymes in different parts of rat gastrointestinal tract.

The aim of this study was to assess the intensity of oxidative stress by measuring levels of lipid peroxidation products in the duodenum, jejunum and colon of rats infected with Hymenolepis diminuta and evaluate the effectiveness of protection against oxidative stress by measuring the glutathione levels and activity of anti-oxidant enzymes: superoxide dismutase, catalase, glutathione reductase and glutathione peroxidase. In exposed rats we observed a significant increase of lipid peroxidation products in the duodenum and jejunum. A significant decrease in superoxide dismutase activity in all the examined parts of the digestive tract was observed. Additionally, rats from 16 to 40 days post H. diminuta infection (dpi) had a decreased catalase activity in the colon, while at 60dpi it increased. The glutathione peroxidase activity increased significantly in the colon at 60dpi. The increase in glutathione reductase activity was observed in the colon in rats 60dpi. There was a lack of changes in the levels of glutathione in the duodenum and a significant increase in its concentration in the jejunum and colon from 40 to 60dpi and from 16 to 40dpi, respectively. In this study we observed altered activity of anti-oxidant enzymes and glutathione level in experimental hymenolepidosis, as a consequence of oxidative stress. It may indicate a decrease in the efficiency of intestinal protection against oxidative stress induced by the presence of the parasite. The imbalance between oxidant and anti-oxidant processes may play a major role in pathology associated with hymenolepidosis.

Tumor-infiltrating lymphocytes in tissue material combined with systemic lymphocyte inflammation in patients with colorectal cancer.

The efficacy of cancer immunotherapy has been actively explored in the treatment of various malignant neoplasms of the gastrointestinal tract. In light of recent reports, the present study aimed to investigate the combination of the absolute lymphocyte count (ALC), percentage of tumor-infiltrating lymphocyte (TILs) and tumor progression status in patients with colorectal cancer (CRC) who underwent surgery. The medical records of 160 patients diagnosed with CRC were retrospectively reviewed. TILs were determined as a percentage of mononuclear inflammatory cells in the total intratumoral or stromal area as determined in five high power fields (magnification, x200-400), at the invasive front and in the centre of the tumour. Blood samples were obtained within 3 days prior to and 7 days following the surgical treatment. The assessment of the TIL percentage was performed in the tissue at the invasive front and in the centre of the primary tumour mass in combination with the determination of ALC in whole blood samples. The samples were obtained prior to and after surgery from patients with CRC, and the tumour progression status was also assessed (TILs/ALC/tumour progression status). A significant association was observed between the percentage of TILs in the main mass of tumour and the tumour size (P=0.031), the pT stage (P=0.049) and the incidence of necrosis (P=0.037) following surgery. The histological type was associated with the evaluated combined parameters prior to surgery (P=0.046). Lymph node pouch invasion was associated with TILs at the invasive front of tumour and with ALC prior to and after surgery (P=0.006 and P=0.037). Furthermore, the data indicated that the percentage of TILs located on the invasive front and centre of the tumour, and the ALC prior to and following surgery correlated with the treatment status (P=0.032, P=0.018, P≤0.001 and P≤0.001). A significant association was noted between eight features and evaluated combined parameters following surgery. These included the tumour size (P=0.021), TNM stage (P<0.001), tumour deposits (P=0.001), incidence of necrosis (P=0.042) and lymph node metastasis (P<0.001). Furthermore, the degree of invasion of venous (P<0.001), lymphatic (P<0.001) and perineural (P<0.001) sites was also significantly associated with TILs, ALC obtained after surgical treatment and tumor progression status. The data demonstrated that local and systemic chronic inflammation was associated with tumour progression in patients with CRC.

Lipoxygenase-derived hydroxyeicosatetraenoic acids--novel perioperative markers of early post-transplant allograft function?

BACKGROUND: Active metabolites of arachidonic acid (AA), eicosanoids, strongly influence renal homeostasis. The aims of this study were to measure perioperative variations in lipoxygenase (LOX)-derived 5-, 12- and 15-hydroxyeicosatetraenoic (HETE) acids levels, and to examine whether (i) dynamics of these eicosanoid generation changes during the first 5 min of renal allograft reperfusion, (ii) examined HETE acids may influence perioperative 20-HETE generation, and (iii) LOX HETE may serve as perioperative markers of early post-transplant allograft function. METHODS: Sixty-nine kidney recipients were divided into early, slow and delayed graft function (EGF, SGF and DGF, respectively) groups. Blood was taken directly before, and in the consecutive minutes of graft reperfusion. HETE concentrations were measured using liquid chromatography. Creatinine levels were measured during the perioperative period, as well as during follow-up visits (first post-transplant year). RESULTS: Our results demonstrated significant differences in the concentrations and dynamics of HETE changes between the examined groups. Moreover, observed changes in HETE concentrations were strongly associated with post-transplant graft function and perioperative 20-HETE synthesis. Application of cut-off limits for newly introduced markers, that is 71.72 ng/mL for 5-HETE(5), 12.3 ng/mL for 12-HETE△(5-0) and -6.1 ng/mL for 15-HETE△(5-0), resulted in 72.5-81.5% sensitivity and 50-54% specificity for SGF/DGF prediction. Moreover, mixed model analysis revealed that recipients classified according to results of 5-HETE(5) and 15-HETE△(5-0) significantly differ in 1-year post-transplant allograft function (P = 0.03 and P < 0.05, respectively), however, not in the frequency of acute rejections' episodes (P = 0.91 and P = 0.31, respectively). CONCLUSION: We hereby report that human kidney transplantations are accompanied by significant changes in LOX AA metabolism, which strongly influences and predicts early (1 year) post-transplant graft function.

[Xanthinuria type I as the cause of nephrolithiasis in 17-years old girl]. / Ksantynuria typu I jako przyczyna kamicy nerkowej u 17-letniej dziewczynki.

Classic xanthinuria is a rare metabolic defect concerning the final reactions of purine catabolism. There are two types of the disorder: type I results from xanthine dehydrogenase (XDH) deficiency, while type II is characterized by lack of both XDH and aldehyde oxidase activity. Both types are clinically similar and are characterized by elevated xanthine concentration in body fluids that can lead to xanthine crystallisation. The most common manifestation of the disease is urolithiasis, but in most cases xanthinuria remains asymptomatic and the diagnosis is accidental. In the paper we report the first case study of xanthinuria in Poland in a child presenting with urolithiasis. 17-years old female patient was diagnosed because of recurrent urinary lithiasis and hypouricemia was detected during routine tests. Plasma and urine concentrations of oxypurines were measured by high-performance liquid chromatography (HPLC) and showed typical features of xanthinuria: hypouricemia, hypouricosuria, xanthinuria and elevated plasma xanthine. The allopurinol loading test demonstrated type I xanthinuria. The presented case report supports that first symptoms of xanthinuria can appear at any age and this disorder should be considered during diagnosing urolithiasis.