Urine Specimens from Pregnant and Nonpregnant Women Inhibitory to Amplification of \u3cem\u3eChlamydia trachomatis\u3c/em\u3e Nucleic Acid by PCR, Ligase Chain Reaction, and Transcription-Mediated Amplification: Identification of Urinary Substances Associated with Inhibition and Removal of Inhibitory Activity

The presence of endogenous amplification inhibitors in urine may produce false-negative results for the detection of Chlamydia trachomatis nucleic acids by tests such as PCR, ligase chain reaction (LCR), and transcription-mediated amplification (TMA). Consecutive urine specimens from 101 pregnant women and 287 nonpregnant women submitted for urinalysis were processed for C. trachomatis detection. Aliquots were spiked with the equivalent of one C. trachomatis elementary body and were tested by three commercial assays: AMPLICOR CT/NG, Chlamydia LCX, and Chlamydia TMA. The prevalence of inhibitors resulting in complete inhibition of amplification was 4.9% for PCR, 2.6% for LCR, and 7.5% for TMA. In addition, all three assays were partially inhibited by additional urine specimens. Only PCR was more often inhibited by urine from pregnant women than by urine from nonpregnant women (9.9 versus 3.1%; P = 0.011). A complete urinalysis including dipstick and a microscopic examination was performed. Logistic regression analysis revealed that the following substances were associated with amplification inhibition: beta-human chorionic gonadotropin (odd ratio [OR], 3.3) and crystals (OR, 3.3) for PCR, nitrites for LCR (OR, 14.4), and hemoglobin (OR, 3.3), nitrites (OR, 3.3), and crystals (OR, 3.3) for TMA. Aliquots of each inhibitory urine specimen were stored at 4 and -70°C and a dilution of 1:10 (84% for PCR, 100% for LCR, and 92% for TMA). Five urine specimens (three for PCR and two for TMA) required phenol-chloroform extraction to remove inhibitors. The results indicate that the prevalence of nucleic acid amplification inhibitors in female urine is different for each technology, that this prevalence may be predicted by the presence of urinary factors, and that storage and dilution remove most of the inhibitors

Simultaneous Determination of Nitrite and Nitrate in Milk Samples by Ion Chromatography Method and Estimation of Dietary Intake

The presence of nitrate and nitrite in foods may be considered hazardous after ingestion in the gastrointestinal tract due to their reaction with naturally occurred secondary amines to form potentially carcinogenic nitrosamines. Due to this fact, a new method was developed in this study for the simultaneous determination of nitrite and nitrate in milk samples using by ion chromatography. Proposed mobile phase composed of sodium hydrogen carbonate and sodium carbonate (1.0 and 3.2 mmol/L) with a flow rate of 0.7 ml/min. The average recoveries for nitrate and nitrite were higher than 86 and 88, respectively. The limit of detection for nitrate and nitrite were 0.24 and 0.09 mg/L, respectively. The results of 102 real milk samples showed nitrate was found in all of the samples (100) with a mean of 34 ± 11 mg/L, while nitrite was found in none of the samples. The mean intake of nitrate in all age groups was lower than World Health Organization guideline. The present assessment concludes that the maximum contaminant level was equal to 82.8 mg/L nitrate. This method was fast, sensitive and accurate and is capable of being an alternative method in food control laboratories for investigation of nitrite and nitrate content. This is the first study of the determination and survey of nitrite and nitrate and exposure assessment of the Iranian population to nitrite and nitrate level in milk, which was widely used in infants and adolescents as one of the basic food components. Copyright © Taylor & Francis Group, LLC

Monitoring the Content of Nitrates in Vegetables and the Influence of the Pickling Technology on the Denitrification Process

The aim of the work was to determine the concentration of nitrites in vegetable products (tomatoes, cucumbers, white cabbage, table beet, carrot, potatoes, onion and green onion, lettuce, spinach and parsley), realized at markets of the cities Ternopil, Kamianets-Podilskyi and Chernivtsi (Ukraine), to separate the distribution of nitrates in vegetables and also to study the influence of lactic microflora on the nitrate content at pickling tomatoes. It was established, that vegetables with the maximum exceed of maximum permissible concentration (MPC) by the nitrate content up to 1,6 times for products of closed soil are realized at markets. For open soil MPC exceed was in average 2,1 times. It was revealed, that most realized samples of tomatoes and leaf salad vegetables have the over-normative exceed of nitrates up to 35 %, and onion – the least one – 20 %. It was established, that nitrates accumulate in different parts of a fruit. In cucumbers, carrot, potato and table beet, the least quantity of nitrates accumulate in the external part of vegetables (near the surface), and the most one – in the central part. At the same time in cabbage and tomatoes, on the contrary, the least quantity – in the central part, the most one – in the area near the base of vegetables (stump). It was established, that at pickling tomatoes with the nitrate content within MPC lactic fermentation takes place with the intensive growth of titrated acidity, the decrease of the nitrate content takes place at this process. Under conditions of pickling tomatoes with the nitrate content two times more than MPC, the pickling process is a bit decelerated, but the nitrate content decreases to the safe level in a finished product. It was established, that vegetables with the nitrates quantity within 1500 mg/kg and more cannot be used in the pickling technology because of the bacteriological influence of nitrates on lactic microflora. Vegetables with such nitrate content must be obligatory condemned

Evidence that the degree of band 3 phosphorylation modulates human erythrocytes nitric oxide efflux – in vitro model of hyperfibrinogenemia

© 2011 – IOS Press and the authors. All rights reservedRecent evidence has shown that plasma fibrinogen, a major cardiovascular risk factor, interacts with the erythrocyte membrane and acts to influence blood flow via erythrocyte nitric oxide (NO) modulation. In the present pioneer in-vitro study, whole blood samples were harvested from healthy subjects and aliquots were incubated in the absence (control aliquots) and presence of fibrinogen at different degrees of band 3 phosphorylation, and the levels of NO, nitrite, nitrate and S-nitroglutathione (GSNO) were determined. Hyperfibrinogenemia interferes with erythrocyte NO mobilization without changing its efflux in a way that seems to be dependent of the degree of band 3 phosphorylation. In presence of higher fibrinogen concentrations the NO efflux is reinforced when band 3 is phosphorylated (p < 0.001). Higher levels of nitrite, nitrate and GSNO were documented (p < 0.05). However, the mechanisms by which fibrinogen signalling modulates erythrocyte function remain to be clarified and are currently under study. These conditions may be considered an approach to be followed in blood storage for transfusions.This study was supported by grants from the FCT - Fundação para a Ciência e a Tecnologia (project reference PTDC/SAU-OSM/73449/2006

Antioxidant and Anti-Inflammatory Activities of Flavanones from Glycyrrhiza glabra L. (licorice) Leaf Phytocomplexes: Identification of Licoflavanone as a Modulator of NF-kB/MAPK Pathway

Inflammation represents an adaptive response generated by injuries or harmful stimuli. Natural remedies represent an interesting alternative to traditional therapies, involving several biochemical pathways. Besides, the valorization of agrochemical wastes nowadays seems to be a feasible way to reduce the health spending and improve the accessibility at bioactive natural compounds. In this context, the chemical composition of three Glycyrrhiza glabra L. (licorice) leaf extracts, obtained through maceration or ultrasound-assisted method (fresh and dried leaves) was investigated. A guided fractionation obtained three main components: pinocembrin, glabranin and licoflavanone. All the extracts showed similar antioxidant properties, evaluated by 2,2'-diphenyl-1-picrylhydrazyl (DPPH) or 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) Diammonium Salt (ABTS) assay, while, among the isolated compounds, licoflavanone exhibited the best antioxidant activity. The anti-inflammatory activity of the extracts and the purified compounds was investigated in lipopolysaccharide (LPS)-stimulated RAW 264.7 murine macrophages. Extract C and licoflavanone showed a good anti-inflammatory activity without affecting cell viability, as they decreased nitrite levels even when used at 12.5 μg/mL (p < 0.005) and 50 μM concentration (p < 0.001), respectively. Interestingly, licoflavanone markedly decreased pro-inflammatory cytokines and cyclooxygenase 2/inducible nitric oxide synthase (COX-2/iNOS) expression levels (p < 0.001). A modulation of nuclear factor kappa B/mitogen-activated protein kinases (NF-kB/MAPK) pathway underlay such behavior, highlighting the potential of this natural compound as a new scaffold in anti-inflammatory drug research

Physico-chemical factors and bacteria in fish ponds

Analyses of pond water and mud samples show that nitrifying bacteria (including ammonifying bacteria, nitrite bacteria, nitrobacteria and denitrifying bacteria) are in general closely correlated with various physico-chemical factors, ammonifying bacteria are mainly correlated with dissolved oxygen; denitrifying bacteria are inversely correlated with phosphorus; nitrite bacteria are closely correlated with nitrites, nitrobacteria are inversely correlated with ammoniac nitrogen. The nitrifying bacteria are more closely correlated with heterotrophic bacteria. Nitrobacteria are inversely correlated with anaerobic heterotrophic bacteria. The correlation is quite weak between all the nitrite bacteria which indicates that the nitrite bacteria have a controlling and regulating function in water quality and there is no interdependence as each plays a role of its own. The paper also discusses how the superficial soil (pond mud down to 3.5 cm deep) and different layers of the mud affect the biomass of bacteria. The study shows that the top superficial layer (down to 1.5 cm deep) is the major area for decomposing and converting organic matter

Effect of operational parameters and Pd/In catalyst in the reduction of nitrate using copper electrode

Water with high concentration of nitrate may cause damage to health and to the environment. This study investigated how concentration, current density, flow, pH, the use of Pd/In catalyst and operating mode (constant current density and constant cell potential) have an influence in the electrochemical reduction of nitrate and in the formation of gaseous compounds using copper electrode. Experiments were performed in two-compartment electrolytic cells separated by a cationic membrane with nitrate model solutions prepared as a surrogate of concentrated brines from membrane desalination plants. The results show that the electroreduction process has potential for reduction of nitrate and that it is influenced by the operational conditions. The best conditions found for the treatment - with satisfactory reduction of nitrate, formation of compounds and reproducibility - were at nitrate concentrations of 600 and 1000 mg.L-1, current density of 1.1 mA.cm-2 and without pH control, since in these conditions the production of gaseous compounds is higher than the production of nitrite. When Pd/In catalyst was used, the nitrate reduction was 50% after 6 hours of experiment and the predominant product were gaseous compounds. When compared to the experiment without the catalyst, the arrangement with Pd/In was the most efficient one.Fil: Beltrame, Thiago Favarini. Ufrgds Lacor; BrasilFil: Coelho, Vanessa. Ufrgds Lacor; BrasilFil: Marder, Luciano. Ufrgds Lacor; BrasilFil: Ferreira, Jane Zoppas. Ufrgds Lacor; BrasilFil: Marchesini, Fernanda Albana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Investigaciones en Catálisis y Petroquímica ; ArgentinaFil: Bernardes, Andrea Moura. Ufrgds Lacor; Brasi

Hyperoxia Causes Mitochondrial Fragmentation in Pulmonary Endothelial Cells by Increasing Expression of Pro-Fission Proteins

Objective—We explored mechanisms that alter mitochondrial structure and function in pulmonary endothelial cells (PEC) function after hyperoxia. Approach and Results—Mitochondrial structures of PECs exposed to hyperoxia or normoxia were visualized and mitochondrial fragmentation quantified. Expression of pro-fission or fusion proteins or autophagy-related proteins were assessed by Western blot. Mitochondrial oxidative state was determined using mito-roGFP. Tetramethylrhodamine methyl ester estimated mitochondrial polarization in treatment groups. The role of mitochondrially derived reactive oxygen species in mt-fragmentation was investigated with mito-TEMPOL and mitochondrial DNA (mtDNA) damage studied by using ENDO III (mt-tat-endonuclease III), a protein that repairs mDNA damage. Drp-1 (dynamin-related protein 1) was overexpressed or silenced to test the role of this protein in cell survival or transwell resistance. Hyperoxia increased fragmentation of PEC mitochondria in a time-dependent manner through 48 hours of exposure. Hyperoxic PECs exhibited increased phosphorylation of Drp-1 (serine 616), decreases in Mfn1 (mitofusion protein 1), but increases in OPA-1 (optic atrophy 1). Pro-autophagy proteins p62 (LC3 adapter–binding protein SQSTM1/p62), PINK-1 (PTEN-induced putative kinase 1), and LC3B (microtubule-associated protein 1A/1B-light chain 3) were increased. Returning cells to normoxia for 24 hours reversed the increased mt-fragmentation and changes in expression of pro-fission proteins. Hyperoxia-induced changes in mitochondrial structure or cell survival were mitigated by antioxidants mito-TEMPOL, Drp-1 silencing, or inhibition or protection by the mitochondrial endonuclease ENDO III. Hyperoxia induced oxidation and mitochondrial depolarization and impaired transwell resistance. Decrease in resistance was mitigated by mito-TEMPOL or ENDO III and reproduced by overexpression of Drp-1. Conclusions—Because hyperoxia evoked mt-fragmentation, cell survival and transwell resistance are prevented by ENDO III and mito-TEMPOL and Drp-1 silencing, and these data link hyperoxia-induced mt-DNA damage, Drp-1 expression, mt-fragmentation, and PEC dysfunction