Gal-3 Deficiency Suppresses Novosphyngobium aromaticivorans Inflammasome Activation and IL-17 Driven Autoimmune Cholangitis in Mice.

Gal-3 has the role in multiple inflammatory pathways. Multiple-hit etiology of primary biliary cholangitis (PBC) and evolving immune response at various stages of the disease includes involvement of Gal-3 in PBC pathogenesis. In this study we aimed to clarify the role of Gal-3 in Novosphingobium aromaticivorans (N. aromaticivorans) induced biliary disease. Autoimmune cholangitis was induced in mice by two intra-peritoneal injections of N. aromaticivorans within 2 weeks. The role of Gal-3 was evaluated by using Lgals3-/- mice and mice treated with Gal-3 inhibitor. The histological and serological parameters of disease, phenotype of dendritic, NK, NKT, and T cells and inflammasome expression were evaluated. Marked attenuation of the disease in Lgals3-/- and Gal-3 inhibitor, DAVANAT®, treated mice is manifested by the absence of bile duct damage, granulomas and fibrosis. Liver infiltrates of N. aromaticivorans infected wild type mice had higher incidence of pro-inflammatory macrophages, dendritic cells, NK, NKT, and T cells. Lgals3 deletion and treatment with Gal-3 inhibitor reduced inflammatory mononuclear cell infiltrate, expression of NLRP3 inflammasome in the liver infiltrates and interleukin-1ß (IL-1ß) production in the livers of N. aromaticivorans infected mice. In vitro stimulation of wild type peritoneal macrophages with N. aromaticivorans caused increased NLRP3 expression, caspase-1 activity and IL-1ß production compared with Lgals3-/- cells. Our data highlight the importance of Gal-3 in promotion of inflammation in N. aromaticivorans induced PBC by enhancing the activation of NLRP3 inflammasome and production of IL-1ß and indicate Gal-3 as possible therapeutical target in autoimmune cholangitis. Galectin-3 appears involved in inflammatory response to gut commensal leading to PBC.

Synthesis, cytotoxic activity and DNA interaction studies of new dinuclear platinum(ii) complexes with an aromatic 1,5-naphthyridine bridging ligand: DNA binding mode of polynuclear platinum(ii) complexes in relation to the complex structure.

The synthesis, spectroscopic characterization, cytotoxic activity and DNA binding evaluation of seven new dinuclear platinum(ii) complexes Pt1-Pt7, with the general formula [{Pt(L)Cl}2(µ-1,5-nphe)](ClO4)2 (1,5-nphe is 1,5-naphthyridine; while L is two ammines (Pt1) or one bidentate coordinated diamine: ethylenediamine (Pt2), (±)-1,2-propylenediamine (Pt3), trans-(±)-1,2-diaminocyclohexane (Pt4), 1,3-propylenediamine (Pt5), 2,2-dimethyl-1,3-propylenediamine (Pt6), and 1,3-pentanediamine (Pt7)), were reported. In vitro cytotoxic activity of these complexes was evaluated against three tumor cell lines, murine colon carcinoma (CT26), murine mammary carcinoma (4T1) and murine lung cancer (LLC1) and two normal cell lines, murine mesenchymal stem cells (MSC) and human fibroblast (MRC-5) cells. The results of the MTT assay indicate that all investigated complexes have almost no cytotoxic effects on 4T1 and very low cytotoxicity toward LLC1 cell lines. In contrast to the effects on LLC1 and 4T1 cells, complexes Pt1 and Pt2 had significant cytotoxic activity toward CT26 cells. Complex Pt1 had a much lower IC50 value for activity on CT26 cells compared with cisplatin. In comparison with cisplatin, all dinuclear Pt1-Pt7 complexes showed lower cytotoxicity toward normal MSC and MRC-5 cells. In order to measure the amount of platinum(ii) complexes taken up by the cells, we quantified the cellular platinum content using inductively coupled plasma mass spectrometry (ICP-QMS). Molecular docking studies performed to evaluate the potential binding mode of dinuclear platinum(ii) complexes Pt1-Pt7 and their aqua derivatives W1-W7, respectively, at the double stranded DNA showed that groove spanning and backbone tracking are the most stable binding modes.

Characterization of gelation process and drug release profile of thermosensitive liquid lecithin/poloxamer 407 based gels as carriers for percutaneous delivery of ibuprofen.

Suitability of liquid lecithin (i.e., solution of lecithin in soy bean oil with ∼ 60% w/w of phospholipids) for formation of gels, upon addition of water solution of poloxamer 407, was investigated, and formulated systems were evaluated as carriers for percutaneous delivery of ibuprofen. Formulation study of pseudo-ternary system liquid lecithin/poloxamer 407/water at constant liquid lecithin/poloxamer 407 mass ratio (2.0) revealed that minimum concentrations of liquid lecithin and poloxamer 407 required for formation of gel like systems were 15.75% w/w and 13.13% w/w, respectively, while the maximum content of water was 60.62% w/w. The systems comprising water concentrations in a range from 55 to 60.62% w/w were soft semisolids suitable for topical application, and they were selected for physicochemical and biopharmaceutical evaluation. Analysis of conductivity results and light microscopy examination revealed that investigated systems were water dilutable dispersions of spherical oligolamellar associates of phospholipids and triglyceride molecules in the copolymer water solution. Rheological behavior evaluation results indicated that the investigated gels were thermosensitive shear thinning systems. Ibuprofen (5% w/w) was incorporated by dispersing into the previously prepared carriers. Drug-loaded systems were physically stable at storage temperature from 5 ± 3°C to 40 ± 2°C, for 30 days. In vitro ibuprofen release was in accordance with the Higuchi model (rH>0.95) and sustained for 12h. The obtained results implicated that formulated LLPBGs, optimized regarding drug release and organoleptic properties, represent promising carriers for sustained percutaneous drug delivery of poorly soluble drugs.

Natural and anthropogenic factors affecting the groundwater quality in Serbia.

Various chemometric techniques were used to analyze the quality of groundwater data sets. Seventeen water quality parameters: the cations Na, K, Ca, Mg, the anions Cl, SO4, NO3, HCO3 and nine trace elements Pb, As, Mn, Ni, Cu, Cd, Fe, Zn and Cr were measured at 66 different key sampling sites in ten representative areas (low land-Northern Autonomous Province of Serbia, Vojvodina and central Serbia) for the summer period of 2009. HCA grouped the sample sites into four clusters based on the similarities of the characteristics of the groundwater quality. DA showed two parameters, HCO3 and Zn, affording more than 90% correct assignments in the spatial analysis of four/three different regions in Serbia. Factor analysis was applied on the log-transformed data sets and allowed the identification of a reduced number of factors with hydrochemical meaning. The results showed severe pollution with Mn, As, NO3, Ni, Pb whereby anthropogenic origin of these contaminants was indicated. The pollution comes from both scattered point sources (industrial and urban effluent) and diffuse source agricultural activity. These samples may not be suitable for human consumption; the water quality belongs to class III/IV (contaminated). The Fe anomalies (7.1mg/L) in the water from the Vetrnica site can be attributed to natural sources, such as the dissolution of rock masses and rock fragments. The serious groundwater contamination with As (25.7-137.8 µg/L) in the area of Banat (Northern Autonomous Province of Serbia, Vojvodina) and a sample No. 9 at the Great Morava River requires urgent attention.

Risk factors associated with carotid artery puncture following landmark-guided internal jugular vein cannulation attempts.

OBJECTIVE: The relationship between certain risk factors and carotid artery puncture (CAP) as an early mechanical complication following internal jugular vein cannulation attempts (IJVCAs) was evaluated. METHODS: In a retrospective 1-year observational single-center study, 86 IJVCAs conducted in the operating room by 4 competent anesthesiologists were evaluated. Age, gender, puncture side, number of cannulation attempts, circumstances of the procedure and incidence of CAP were obtained from medical records. RESULTS: Of the 86 IJVCAs performed in patients aged 18-75 years, CAP occurred in 8 (9.3%): 5 (5.8%) in patients >65 years and 3 (3.5%) in patients <65 years of age. CAP was not associated with patient's age (p = 0.11) and gender (p = 0.76). Multiple cannulation attempts (OR = 26.25; 95% CI = 4.52-152.51; p < 0.001) and placement of CVC under emergency conditions (OR = 14.84; 95% CI = 1.73-127.22; p = 0.014) increased the risk for CAP significantly. Also, the risk for CAP was higher when IJVCAs were performed before induction of general anesthesia (OR = 15.75; 95% CI = 1.83-135.1; p = 0.019). CAP was more likely to happen during left-sided than right-sided IJVCA (OR = 5.98; 95% CI = 1.29-27.59; p = 0.022). In addition, left-sided attempts considerably increased the risk for multiple cannulation attempts (OR = 2.782; 95% CI = 1.342-3.965; p < 0.01). Also, manifold cannulation attempts were more frequent if the IJVCA was performed before induction of anesthesia (OR = 4.219; CI = 1.579-11.271; p = 0.004). CONCLUSIONS: Our results strongly suggest that left-sided, multiple IJVCAs, performed under emergency conditions in conscious patients in the operating room, represent considerable risks for possible CAP.

[Rapid simultaneous determination of organophosphorus pesticides in human serum and urine by liquid chromatography-mass spectrometry].

BACKGROUND/AIM: Analysis of organophosphosphorus compounds and their metabolites in a biological material includes the use of numerous methods, covering both preparation of samples for analysis and their identification that is considered to be very complex. Low concentrations monitoring requires implementation of highly sensitive analytical techniques. The aim of this study was to develop and validate an original and sensitive method for the detection and quantitation of organophosphorus pesticides (dimethoate, diazinon, malathion and malaoxon) in human biological matrices (serum, urine). METHODS: This method was based on a solid-phase extraction procedure, a chromatographic separation using an ACQUITY UPLC HSST3 column and mass spectrometric detection in the positive ion mode. Mobile phase: was consited of Solvent A (5 mM ammonium formate pH 3.0) and Solvent B (0.1% acetic formate in methanol), in a linear gradient (constant flow-rate 0.3 mL/min). RESULTS: The standard curve was linear in the range of 0.05-5.00 mg/L for malathion and malaoxon, 0.10-5.00 mg/L for dimethoate and 0.05-2.50 mg/L for diazinon. The correlation coefficient was r > or = 0.99. Extraction recoveries were satisfactory and ranged between 90-99%. The limits of detection (LOD) was between 0.007-0.07 mg/L and the limits of quantitation (LOQ) ranged between 0.022-0.085 mg/L. Intra- and interassay precision and accuracy were satisfactory for all of the pesticides analyzed. CONCLUSION: The method of liquid chromatography-mass spectrometry is simple, accurate, and useful for the determination of organophosphorus pesticides in both clinical and forensic toxicology.

Liver, gills, and skin histopathology and heavy metal content of the Danube sterlet (Acipenser ruthenus Linnaeus, 1758).

The sterlet (Acipenser ruthenus L.) is a bottom-feeding fish species with a direct exposure to contaminants from water and sediments. Although heavy metal pollution is believed to be one of the main threats to the sterlet population in the Danube River basin, there is a lack of knowledge of the exact impact of heavy metals on their survival. In the present study, effects of heavy metal pollution on sterlet in the Danube basin were assessed as well as the utility of different sterlet organs and tissues as indicators of heavy metal contamination. The sterlet were sampled at three different sites in the Danube basin, in Hungary and Serbia, isolated from each other by dams. Heavy metal analysis included measurement of Cd, As, Pb, Cr, Hg, Cu, Ni, Fe, Mn, and Zn concentrations in sterlet gills, muscle, liver, and intestine, and histopathological analyses comprised assessment and scoring of the extent and intensity of alterations in skin, gills, and liver tissue. Analysis revealed a significant presence of sublethal histopathological changes that were most pronounced in the liver and skin and increased accumulation of heavy metals, with the highest concentrations in the liver. Canonical discriminant analysis showed significant differentiation among the three studied localities, suggesting that the heavy metal concentrations in sterlet populations were site specific. The present study concludes that the accumulation of heavy metals is a response to the presence of these pollutants in the environment, and, together with other pollutants, it affects the vital organs of natural sterlet populations.

[Interrelation between well-being of the mother and heart rate variability of her preterm infant]. / Zusammenhänge zwischen dem Wohlbefinden der Mutter und der Herzfrequenzvariabilität von Frühgeborenen.

The relationship between mothers' well-being and the heart rate variability of their preterm babies was investigated. In order to study a possible influence of the mother's well-being on the calming quality of her voice and thereby on the heart rate variability of her preterm infant, maternal/paternal stress and competences as well as family functionality were assessed via respective questionnaires. (N = 30) Preterm babies at the postnatal age of approximately 4 weeks were acoustically stimulated with the voice of their own mother. Various heart rate variability measures (NN interval mean value, NN interval median, variance of NN intervals, standard deviation of NN intervals, pnn 6,25, RMSSD, SDSD and RSA) were recorded 15 minutes before, 15 minutes during and 15 minutes after the acoustic stimulation. Non-REM sleep sections of 2 minutes duration were matter of analyses. The correlations between the mothers' well-being and their babies' heart rate variability indicate a strong relationship. The correlations point out that a higher family functionality is associated with a higher heart rate variability of preterm babies. Contradictory to the expectations, higher burden and lower resources as well as lower competences of the mothers were associated with a higher heart rate variability of the preterm babies. Simultaneous real-time investigations of the mothers' and the babies' heart rate variability during a live mother-baby-interaction seems necessary to provide further explanations.