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Precision Spectroscopy on OHFast, Arthur 27 May 2019 (has links)
No description available.
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The Investigation of Photocatalysts and Iron Based Materials in the Oxidation and the Adsorption of Toxic Organic and Chromium MaterialsJiang, Wenjun 13 November 2013 (has links)
The presences of heavy metals, organic contaminants and natural toxins in natural water bodies pose a serious threat to the environment and the health of living organisms. Therefore, there is a critical need to identify sustainable and environmentally friendly water treatment processes. In this dissertation, I focus on the fundamental studies of advanced oxidation processes and magnetic nano-materials as promising new technologies for water treatments.
Advanced oxidation processes employ reactive oxygen species (ROS) which can lead to the mineralization of a number of pollutants and toxins. The rates of formation, steady-state concentrations, and kinetic parameters of hydroxyl radical and singlet oxygen produced by various TiO2 photocatalysts under UV or visible irradiations were measured using selective chemical probes. Hydroxyl radical is the dominant ROS, and its generation is dependent on experimental conditions. The optimal condition for generation of hydroxyl radical by of TiO2 coated glass microspheres is studied by response surface methodology, and the optimal conditions are applied for the degradation of dimethyl phthalate. Singlet oxygen (1O2) also plays an important role for advanced processes, so the degradation of microcystin-LR by rose bengal, an 1O2 sensitizer was studied. The measured bimolecular reaction rate constant between MC-LR and 1O2 is ~ 106 M-1 s-1 based on competition kinetics with furfuryl alcohol.
The typical adsorbent needs separation after the treatment, while magnetic iron oxides can be easily removed by a magnetic field. Maghemite and humic acid coated magnetite (HA-Fe3O4) were synthesized, characterized and applied for chromium(VI) removal. The adsorption of chromium(VI) by maghemite and HA-Fe3O4 follow a pseudo-second-order kinetic process. The adsorption of chromium(VI) by maghemite is accurately modeled using adsorption isotherms, and solution pH and presence of humic acid influence adsorption. Humic acid coated magnetite can adsorb and reduce chromium(VI) to non-toxic chromium (III), and the reaction is not highly dependent on solution pH. The functional groups associated with humic acid act as ligands lead to the Cr(III) complex via a coupled reduction-complexation mechanism. Extended X-ray absorption fine structure spectroscopy demonstrates the Cr(III) in the Cr-loaded HA-Fe3O4 materials has six neighboring oxygen atoms in an octahedral geometry with average bond lengths of 1.98 Å.
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Sources of 2,5-Diaminoimidazolone Lesions in DNA Damage Initiated by Hydroxyl Radical AttackThomas, Caroline Suzanne, Pollard, Hannah Catherine, Razskazovskiy, Yuriy, Roginskaya, Marina 02 July 2020 (has links)
The present study reports radiation-chemical yields of 2.5-diaminoimidazolone (Iz) derivatives in X-irradiated phosphate-buffered solutions of guanosine and double-stranded DNA. Various gassing conditions (air, N20/O2 (4:1), N2O, vacuum) were employed to elucidate the contribution of several alternative pathways leading to Iz in reactions initiated by hydroxyl radical attack on guanine. In all systems, Iz was identified as the second by abundance guanine degradation product after 8-oxoguanine, formed in 1:5 (guanosine) and 1:3.3 (DNA) ratio to the latter in air-saturated solutions. Experimental data strongly suggest that the addition of molecular oxygen to the neutral guanine radical G(-H)• plays a major in Iz production in oxygenated solutions of double-stranded DNA while in other systems it may compete with recombination of G(-H)• with superoxide and/or alkyl peroxyl radicals. The production of Iz through hydroxyl radical attack on 8-oxoguanine was also shown to take place although the chemical yield of Iz (ca 6%) in this process is too low to compete with the other pathways. The linearity of Iz accumulation with dose also indicates a negligible contribution of this channel to its yield in all systems.
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Hydroxyl Radical Production via Acoustic Cavitation in Ultrasonic Humidifier SystemsAltizer, Chase Duncan 12 June 2018 (has links)
Ultrasonic humidifiers use sound vibrations at frequencies higher than can be heard by humans (> 20,000 Hz) to generate aerosolized water also have potential for inducing sonochemical reactions for chemicals present in water. This research focuses on examining oxidants formed within ultrasonic humidifiers, as well as the oxidants effects of contaminants in water used in the systems. Hydroxyl radicals were found using DMPO as a spin trap. Caffeine and 17β-estradiol, as pharmaceutical contaminants of drinking water, were both emitted from the humidifier when present in the water reservoir and would enter breathing air. Emitted 17β-estradiol was found at 60% of the initial concentration filled in the ultrasonic humidifier after 480 minutes. Caffeine exhibited less degradation than 17β-estradiol. Degradation of both pharmaceuticals was attributed to ultrasonic processes, most likely oxidation with hydroxyl radicals produced. Bromide as a contaminant of the fill water was found to remain constant over time. / MS
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Sol-gel Synthesis and Photocatalytic Characterization of Immobilized TiO2 FilmsLiao, Haidong January 2009 (has links)
<p> Contamination of surface and ground water from industrial wastes and anthropogenic activities represents one of the greatest challenges to the sustainable development of human society. Heterogeneous photocatalysis, a kind of advanced oxidation process characterized by the production of highly oxidative hydroxyl radicals, is a relatively novel subject with tremendous potential in water treatment applications.</p><p> The purpose of this research was first to develop feasible hydroxyl radical detection methods, which can be used to evaluate efficiency of photocatalytic process, and second to prepare immobilized TiO2 films with high photocatalytic activities by the sol gel method.</p><p> The feasibility of Indigo carmine and phthalic hydrazide as OH-radical probes was investigated. The organic dye Indigo carmine absorbs visual light strongly at 610 nm and its destruction can be monitored conveniently in a spectrophotometer. Results showed that both <sup>·</sup>OH and HO<sub>2</sub><sup>·</sup><sub> </sub>can bleach Indigo carmine, and the bleaching yield of <sup>·</sup>OH was pH independent. The photocatalytic dye bleaching in black light UV illuminated Degussa P25 TiO2 aerated suspensions was then investigated. A strong pH dependency of the bleaching yield was found. This implies that the quantum yield of OH radical at pH 3 is one fourth compared to that at pH 10. The reaction of the OH radical with phthalic hydrazide will form strongly chemiluminescent 3-hydroxyphthalic hydrazide. Using the more specific phthalic hydrazide as OH radical probe, an even stronger pH dependent quantum yield of OH radical was found. At pH 10 the quantum yield reached the same magnitude as that obtained by using Indigo carmine, whereas the quantum yields at acidic pH were close to zero. However it was found that the addition of phosphate and fluoride anions can substantially enhance the OH radical yield at acidic pH by blocking the adsorption of phthalic hydrazide onto the TiO2 surfaces. Hence the adsorption of phthalic hydrazide to TiO2 is an important factor to consider when this method is used. <em></em></p><p> Photocatalytic TiO2 films coated on metal plates were prepared by a sol gel method using titanium isopropoxide as TiO2 precursor and isopropanol as solvent. The photocatalytic activity of the obtained films was evaluated by bleaching of indigo carmine at pH 9 under black light UV irradiation. The effect of the molar ratio of isopropanol, water and hydrochloric acid to titanium isopropoxide was studied. It was also shown that the activities of TiO2 films are considerably influenced by calcination temperature, coating cycles and the supporting materials.</p> / <p> Förorening av yt- och grundvatten från industrier och humana aktiviteter utgör en av de största utmaningarna för en hållbar utveckling av det mänskliga samhället. Heterogen fotokatalys, en slags avancerad oxidations process som kännetecknas av att starkt oxidativa hydroxylradikaler produceras, är en relativt ny teknik med stor potential för vattenrening.</p><p> Ett syfte med detta licentiatarbete var först att utveckla och genomföra olika metoder för att detektera bildningen av hydroxylradikaler såväl i laboratoriet som i tekniska miljöer. Det andra syftet med arbetet var att syntetisera immobiliserade TiO2 filmer med hög fotokatalytisk effektivitet med en sol-gel metod.</p><p> Möjligheten att använda indigokarmin och ftalhydrazid som OH-radikalprob undersöktes. Det organiska färgämnet indigokarmin absorberar synligt ljus starkt vid 610 nm vilket gör att dess nedbrytning lätt kan följas i en spektrofotometer. Resultaten av gammaradiolys visade att båda <sup>•</sup>OH och HO<sub>2</sub><sup>•</sup> kan bleka indigokarmin och att den blekning som härrör från <sup>•</sup>OH var oberoende av pH. Fotokatalytisk blekning av indigokarmin med blacklight UV bestrålning av Degussa P25 TiO2 suspensioner undersöktes sedan. Ett starkt pH-beroende av blekningsutbytet erhölls vilket tolkas som att kvantutbytet av OH-radikaler vid pH 3 är en fjärdedel jämfört med det vid pH 10.</p><p> När ftalhydrazid reagerar med OH-radikaler bildas starkt kemiluminiscent 3-hydroxy-ftalhydrazid. Med denna specifika OH-radikalprob, erhölls ett ännu starkare pH beroende. Vid pH 10 var kvantutbytet i paritet med det som erhölls med indigokarmin, medan kvantutbytet vid lågt pH var nära noll. Tillsats av fosfat- och fluoridjoner visade sig avsevärt öka OH-radikalutbytet vid lågt pH-värde genom att blockera adsorption av ftalhydrazid på TiO2 ytorna. Adsorptionen av ftalhydrazid på TiO2 är således en viktig faktor när denna metod används.</p><p> Fotokatalytiskt verksamma TiO2-filmer på metallplattor framställdes med sol-gel metoden med titan-isopropoxide som TiO2-prekursor och isopropanol som lösningsmedel. Den fotokatalytiska aktiviteten av TiO2-belagda plattor utvärderades genom blekning av indigokarmin vid pH 9 under blacklight UV-bestrålning. Effekten av olika proportioner mellan isopropanol, vatten och saltsyra till titan isopropoxide undersöktes. Det visade sig att denfotokatalytiska aktiviteten av TiO2-plattorna i hög grad påverkades av kalcineringstemperatur, beläggningscykler och materialet i plattorna.</p>
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Liquid Aerosol PhotochemistryBones, David Lawrence January 2008 (has links)
Aerosols of nitrate solutions were irradiated in the presence of radical scavengers in an attempt to measure the yield of hydroxyl radical in both the aqueous phase and the gas phase. Carbon monoxide, benzoic acid, benzene and cyclohexane were used as scavengers to trap hydroxyl radical. The products from the reaction of these scavengers with hydroxyl radical were analysed with High Performance Liquid Chromatography and mass spectrometry. The radiant flux in the chamber was measured via ferrioxalate actinometry, both with bulk liquid and aerosol droplets. Many quantitative results were obtained but several anomalies were found. This suggests that Mie theory is not capable of predicting rates of photochemical reactions within droplets.
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Preparação e caracterização de fotocatalisadores heterogêneos de titânio e nióbio e avaliação do potencial de fotodegradação / Preparation and characterization of niobium and titanium heterogeneous photocatalyst and evaluation of their potential in photodegradationGallo, Inara Fernanda Lage 06 May 2016 (has links)
É urgente o desenvolvimento de novas tecnologias para o tratamento de água nos dias atuais. Neste contexto, os processos oxidativos avançados (POA) tem sido bem-sucedido no tratamento de contaminantes presentes em efluentes industriais e na rede de esgoto doméstico. Neste trabalho, estudamos um dos POA, a fotocatáliseheterogênea por meio da síntese de fotocatalisadores mistos de nióbio e titânio utilizando-se o método Pechini, com uma temperatura de calcinação de 470 oC. Foram sintetizados os fotocatalisadores PNB000, PNB018, PNB030, PNB070, PNB099 e PNB100, onde os números mostram aporcentagem em mols de pentóxido de nióbio presente em cada um deles. Estes fotocatalisadores foram submetidos a um segundo tratamento térmico a 800oC, durante 6 horas, e obtivemoso PNB000_01, PNB018_01, PNB030_01, PNB070_01 e PNB100_01. A caracterização dos fotocatalisadores foi feita por: análise de espectroscopia de energia dispersiva (EDS); área superficial determinadas por isotermas de adsorção e dessorção de nitrogênio, BET; imagens de microscopiaeletrônica de varredura (MEV); difração de raios-X; análises térmicas (termogravimetria e análise térmica diferencial), determinação de band gap por reflectância difusa. O potencial para serem usados como fotocatalisadores heterogêneos para degradação de compostos orgânicos foi avaliado pela eficiência fotônica de geração de radicais hidroxilas(OH)sob radiação de lâmpada UVA (15 W). A análise de EDS confirmou acomposiçãoem mol de TiO2 e Nb2O5presentes nosfotocatalisadoressintetizados. A área superficial obtida por isotermas (BET) do PNB018 (161,7 m²/g), PNB030 (130,8 m²/g), PNB070 (150,5 m²/g) mostraram-se maiores do que ado TiO2 P25 (52,68 m²/g). As imagens de MEV mostrou que estes fotocatalisadoressão constituídos de partículas manométricas. Outra característica dos óxidos com quantidade intermediária de mols de Nb2O5 (18, 30 e 70%) foi a estrutura amorfa determinada por difração de raios-X. Por sua vez, os óxidos tratados a 800oC apresentaram estruturas cristalinas edifratogramas de raios-X completamente diferentes do TiO2 e do Nb2O5, comprovando-se que são novos materiais.As medidas de energia de band gapmostrou diferenças significativas quando comparamos o TiO2 P25 (Egap 3,22eV) e o PNB000 (Egap 2,90eV). Isso leva a concluir que estes fotocatalisadores sintetizados pelo método Pechini necessitam de uma energia menor para que ocorram as transições eletrônicas. O fotocatalisador PNB070, que apresenta 70% em mols de pentóxido de nióbio, apresentouOH de 0,104 da mesma ordem de grandeza do TiO2 P25 (OH 0,134) e ligeiramente menor do que o OH do Nb2O5.nH2O (OH 0,164). Dessa maneira, supõe-se que o PNB070 tenha o mesmo potencial de eficiência que o TiO2 P25 para fotodegradações. Por sua vez, os fotocatalisadores que passaram por um segundo tratamento térmico a 800 oC mostraram valores de rendimento quântico de produção de radicais hidroxilas bem inferiores aos seus originais. Por exemplo, o PNB070_01 teve OH de 0,003. Esses resultados sugerem que o aumento da cristalinidade pode diminuir a atividade fotocatalítica, ou, que o estado amorfo dos fotocatalisadores deste trabalho favorece o aumento da velocidade de transferência de elétrons e a da fotocatálise. / Nowadays, the development of new technologies for the treatment of water is urgent. In this context, the advanced oxidative processes (AOP) has been successful in the treatment of contaminants foundin industrial effluents and domestic sewage. In this work, we have studied an AOP, the heterogenousphotocatalysis, by means the synthesis of niobium and titanium mixedphotocatalysts, employing calcination temperature of 470 oC.The PNB000, PNB018, PNB030, PNB070, PNB099 and PNB100photocatalysts were synthetized, where the numbers show the percentage in mol of Nb2O5content in each of them. These photocatalytswere submitted to a second heat treatment to 800oC during 6 hours, and PNB000_01, PNB018_01, PNB030_01, PNB070_01 and PNB100_01 were obtained. The characterization of the photocatalysts was made by: analysis of the energy dispersive spectroscopy (EDS); the surface area determined by adsorption and desorption isotherms of nitrogen, BET; images of scanning electron microscopy (SEM); X-ray diffraction; thermal analysis (thermogravimetry and differential thermal analysis), determination of the band gap by diffuse reflectance.Their potential to be used as heterogeneous photocatalystsfor degradation of organic compounds was evaluated by means the determination of the photonic efficiency for hydroxyl radical production (OH)under radiation of UV-A lamp (15 W). The EDS analysis confirmed the composition in mol of TiO2 and Nb2O5 present in the synthesizedphotocatalysts. Surface area obtained by isotherms (BET) of PNB018 (161.7 m ²/g), PNB030 (130.8 m ²/g), PNB070 (150.5 m ²/g) were higher than TiO2 P25 (52.68 m ²/g). SEM images showed that these photocatalysts consist of nanoparticles. Another feature of the oxides with intermediate amount of moles of Nb2O5 (18, 30 and 70%) was the amorphous structure determined by X-ray diffraction. On the other hand, the oxides treated to 800 oC showed crystalline structures and X-ray patternscompletely different from TiO2 and Nb2O5, proving that the synthetized oxides are new materials.The band gap energy measurements showed significant differences when we compare TiO2 P25 (Egap 3.22 eV) and the PNB000 (Egap 2.90 eV). This allow us to conclude that thephotocatalysts synthesized by Pechini method require lower energy in order to have electronic transitions. The PNB070 photocatalyst, which has 70% in mols of niobium pentoxide, showed OHof 0.104 of the same order of magnitude of TiO2 P25 (OH0.134) and slightly smaller than the OH of Nb2O5.nH2O (OH 0.164). For this reason, we can assume that PNB070 has the same potential of the TiO2 P25 for organic compound photodegradation. The photocatalyststhat were submittedthrough a second heat treatment at 800 oC showed quantum efficiency of hydroxyl radical production values well below their original. For instance, PNB070_01 showed OHof 0.003. These results suggest that increased crystallinity can reduce the photocatalytic activity, or the amorphous structures of the photocatalysts of this work improve the electron transfer rates and the photocatalysis.
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Development of an Electrochemical Technique for Oxidative Surface Mapping to Investigate Solution-Phase Protein Dynamics with High Performance Mass Spectrometry and Advanced InformaticsMcClintock, Carlee Suzanne Patterson 01 May 2010 (has links)
Oxidative protein surface mapping has gained popularity over recent years within the mass spectrometry (MS) community for gleaning information about the solvent accessibility of folded protein structures. The hydroxyl radical targets a wide breadth of reactive amino acids with a stable mass tag that withstands subsequent MS analysis. A variety of techniques exist for generating hydroxyl radicals, with most requiring sources of radiation or caustic oxidizing reagents. The purpose of this research was to evaluate the novel use of electrochemistry for accomplishing a comparable probe of protein structure with a more accessible tool. Two different working electrode types were tested across a range of experimental parameters, including voltage, flow rate, and solution electrolyte composition, to affect the extent of oxidation on intact proteins. Results indicated that the boron-doped diamond electrode was most valuable for protein research due to its capacity to produce hydroxyl radicals and its relatively low adsorption profile. Oxidized proteins were collected from the electrochemical cell for intact protein and peptide level MS analysis. Peptide mass spectral data were searched by two different “hybrid” software packages that incorporate de novo elements into a database search to accommodate the challenge of searching for more than forty possible oxidative mass shifts. Preliminary data showed reasonable agreement between amino acid solvent accessibility and the resulting oxidation status of these residues in aqueous solution, while more buried residues were found to be oxidized in “non-native” solution. Later experiments utilized higher flow rates to reduce protein residence time inside the electrochemical flow chamber, along with a different cell activation approach to improve controllability of the intact protein oxidation yield. A multidimensional chromatographic strategy was employed to improve dynamic range for detecting oxidation of lower reactivity residues. Along with increased levels of oxidation around “reactive hotspot” sites, the enhanced sensitivity of these measurements uncovered a significant level of background oxidation in control proteins. While further work is needed to determine the full utility that BDD electrochemistry can lend protein structural studies, the experimental refinements reported here pave the way for improvements that could lead to a high-throughput structural pipeline complementary to predictive modeling efforts.
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Oxygen free radicals : mediators of vascular toneBharadwaj, Lalita Anne 01 January 1997 (has links)
<i>In vivo</i> and <i>in vitro</i> studies on numerous types of blood vessels obtained from a variety of vascular beds and species have demonstrated that oxygen free radicals (OFRs) can evoke both vasodilation and vasoconstriction. Specific OFRs have been shown to elicit different and often times opposite effects on vascular smooth muscle. Therefore, this thesis attempts to define the vascular actions and mechanism of oxygen free radicals (OFRs) [superoxide anion (O<sub>2</sub><sup>-</sup>), hydrogen peroxide (HO<sub>2</sub>) and hydroxyl radical (OH)] on isolated rabbit aorta. This thesis will examine the role of OH in Ach- and nitroglycerin (NTG)-induced relaxation of isolated rabbit aorta. Superoxide anions generated by xanthine (X) plus xanthine oxidase (XO) produced concentration-dependent contractions of isolated rabbit aorta. The contractile response to O<sub>2</sub><sup>-</sup> was completely abolished in preparations denuded of endothellum or pretreated with superoxide dismutase (SOD), a scavenger of O<sub>2</sub><sup>-</sup>. The contractile response was reduced by indomethacin (I), a cyclooxygenase inhibitor. These results suggest that O<sub>2</sub><sup>-</sup> mediated by vasoconstrictor arachidonic acid metabolites. Hydrogen peroxide generated by glucose and glucose oxidase produced contraction (low concentrations) and relaxation followed by contraction (high concentrations) in isolated rabbit aorta. The contractile response was abolished in the presence of catalase, a scavenger of H<sub>2</sub>O<sub>2</sub> however the relaxant effect was exaggerated. Indomethacin markedly reduced the H<sub>2</sub>O<sub>2</sub>-induced contraction. Relaxation was completely prevented by de-endothelialization or pretreatment with N<sup>G</sup>-monomethyl-L-arginine (LNMMA), an inhibitor of nitric oxide synthetase. These results suggest that H<sub>2</sub>O<sub>2</sub> in large concentrations produces a biphasic response, relaxation followed by contraction. Relaxation is endothelium dependent and is mediated by endothelium-derived relaxing factor (EDRF), nitric oxide (NO). The contractile response is endothelium independent and is mediated by vasoconstrictor arachidonic acid metabolites of smooth muscle. Hydroxyl radicals generated by dihydroxyfumarate (DHF), ferric chloride (FeCl<sub>3</sub>) and adenosine diphosphate (ADP) (DHF/FeCl<sub>3</sub>-ADP) produced concentration dependent relaxations of NE-precontracted rabbit aorta. Mannitol (Ml) completely inhibited OH-induced relaxation. Relaxation was markedly reduced in aortic rings mechanically denuded of endothelium. The relaxant effect was reduced by an inhibitor of NO synthesis (LNMMA), by an inhibitor of guanylate cyclase (methylene blue), by an inhibitor of cyclooxygenase (indomethacin) and by an inhibitor of an ATP-sensitive K<sup>+</sup> channel blocker (glyburide). These results indicate that OH produces relaxation that is endothelium-dependent and partially mediated by an endothelium-derived relaxing factor (NO), vasodilatory arachidonic acid metabolites and an ATP-sensitive K<sup>+</sup> channel. We hypothesized that Ach-induced vascular relaxation is mediated by OH derived from the interaction of NO and O<sub>2</sub><sup>-</sup>. To test this hypothesis we investigated the effect of Ach and NTG on NE-precontracted isolated rabbit aortic preparations in the absence or presence of scavengers of O<sub>2</sub><sup>-</sup> (SOD), and OH (dimethylthiourea (DMTU) or mannitol or Garlicin). The OFR scavengers (SOD, dimethylthiourea, mannitol, garlicin and histidine) alone or the combination of SOD and DMTU markedly reduced Ach- or NTG-induced relaxation. However, the combination of histidine, (a <sup>1</sup>O<sub>2</sub> scavenger) SOD and DMTU completely abolished Ach-induced relaxation.
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Histochemical and scanning electron microscopic study of endotoxin-induced changes in vascular endothelial cells and villus goblet cells of rat intestineLiu, Shang-Pin 22 December 2009 (has links)
Intravenous application of a high dose of endotoxin, such as lipopolysaccharide (LPS), results in endotoxemia and sepsis in experimental animals. LPS induces production of cytokines and free radicals, plasma leakage and systemic inflammation. But the relationship between LPS-induced plasma leakage and endothelial gap formation is still unknown. Under normal physiological and pathological conditions, the mucus of intestine plays an important role in host defense mechanism as a barrier to prevent invasion of bacteria and endotoxin. The integrity of the intestinal epithelium is an important determinant of clinical outcome in septic patients. It is reported that, after LPS application, ileal mucosa is injured consequently. Necrosis of epithelial cells is also prominent feature in the villus epithelium. However, the response of mucin-secreting goblet cells is often ignored. The present study was designed to prove (1) whether LPS application increased plasma leakage by endothelial gap formation in rat intestinal tract, (2) whether LPS application increased goblet cell secretion by compound exocytotic activity in mucosal villi of small intestine; and (3) whether hydroxyl radicals were involved in LPS-induced compound exocytosis in goblet cells and plasma leakage.
First, the microcirculation of large intestine in rats was shown by using silver nitrate staining method, and India ink was used to label the leaky microvessels to express the magnitude of plasma leakage. Endothelial gaps formed between endothelial cells in the venules after LPS-induced inflammation were investigated by light and scanning electron microscopy. In saline control, the number of endothelial gaps per 1000 £gm2 endothelium of postcapillary and collecting venules was 0.2 ¡Ó 0.1 ~ 0.4 ¡Ó 0.1 / 1000 £gm2 (n = 5). At 5 minutes after LPS application, the endothelial gap density drastically increased to 12.1 ¡Ó 1.6 ~ 27.5 ¡Ó 2.2 / 1000 £gm2 (n = 5 or 6), about 43-69 times (P < 0.01) as much as control. At the same time, the magnitude of plasma leakage, expressed by area density of India ink-labeled blood vessels, in the cecum and colon of LPS-treated rats increased to 7.8-8.2 times (P < 0.01) as much as control. Unusually high degree of plasma leakage and high number of endothelial gaps persisted for at least 30 minutes after treatment. Then, a significant reduction to the baseline level occurred at 60 minutes after LPS application (P > 0.05). The results evidently indicated that LPS-induced intestinal plasma leakage and the endothelial gap formation of venules were closely related.
In the following experiment, in order to obtain an actual number of goblet cells in the mucosal epithelium, an innovative and effective experimental method was developed and adopted to prepare small intestine specimens in this study. Tissue pieces with two rows of mucosal villi were taken under a dissecting microscope. Then, scanning electron microscope was used to observe goblet cells and histochemistry staining was applied to further identify mucosubstance. The degree of goblet cell secretion in the villus epithelium of the duodenum and ileum was expressed by the number of cavitated goblet cells undergoing compound exocytosis. Digital morphometric software SimplePCI was employed to measure the epithelial surface area of sampled villi and to count the number of goblet cells. In addition, hydroxyl radical scavenger ¡V dimethylthiourea (DMTU) was also applied to explore the role of hydroxyl radicals involving in LPS-induced goblet cells secretion and plasma leakage. From scanning electron microscopy study, the numbers of cavitated goblet cells per mm2 of ileal villus epithelium in rats at 5 and 30 minutes after LPS injection were 693 ¡Ó 196 (n = 6) and 547 ¡Ó 213 (n = 6), respectively, which were 5.1 and 8.4 times (P < 0.05) compared with the number of saline control. The percentage of villus cavitated goblet cell numbers, in both duodenum and ileum 5 minutes after LPS and in the ileum 30 minutes after LPS, increased significantly (P < 0.05). When DMTU was given prior to LPS, the number of cavitated goblet cells and the amount of plasma leakage was inhibited and remained at the level as control (P > 0.05). It is concluded that the mechanism of the LPS-induced increase in compound exocytotic activity of goblet cells and increase in plasma leakage during acute phases of inflammatory response in rat small intestine was associated with hydroxyl radicals.
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