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Mecanismos de sintese e degradação do radical oxido nitrico pela mitocondria de Arabidopsis thaliana / Mechanisms of synthesis and degradation of radical nitric oxide by mitochonderia of Arabidopsis thalianaWulff, Alfredo 14 March 2008 (has links)
Orientador: Ione Salgado / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-11T11:16:49Z (GMT). No. of bitstreams: 1
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Previous issue date: 2008 / Resumo: O objetivo deste trabalho foi estudar os mecanismo s de síntese e de degradação do radical óxido nítrico (NO) pela mitocôndria vegetal, com ênfase à influência das NAD(P)H desidrogenases e da oxidase alternativa (AOX) nestes processos. Os experimentos foram realizados com mitocôndrias isoladas de células em cultura da planta modelo Arabidopsis thaliana. Inicialmente estabeleceu-se um protocolo para o isolamento e purificação de mitocôndrias, cuja integridade funcional foi determinada pelo controle respiratório, potencial elétrico de membrana e transporte eletroforético de cálcio. Os processos de síntese e de degradação de NO foram analisados monitorando-se o consumo de oxigênio e as concentrações de NO no meio de reação, com eletrodos específicos. Os resultados mostraram que o nitrito inibiu a respiração das mitocôndrias de A. thaliana em competição com o oxigênio do meio. Esta inibição foi revertida por cPTIO, um seqüestrador de NO. Quando a AOX foi inibida por n-propilgalato, o efeito inibitório do nitrito na respiração mitocondrial foi aumentado, enquanto que, na presença de myxothiazol, que previne a redução da citocromo c oxidase (COX), o nitrito não teve efeito na respiração. Os efeitos observados do nitrito no consumo de oxigênio sugeriram que NO foi sintetizado pelas mitocôndrias de A. thaliana através de uma atividade redutora de nitrito. Entretanto, a síntese de NO a partir de nitrito foi principalmente detectada em anaerobiose. Em altas concentrações de nitrito (1-5 mM), a produção de NO se mostrou praticamente insensível aos inibidores da cadeia respiratória, enquanto que, em concentrações menores de nitrito (100 -200 ?M) a COX, mas não a AOX, contribuiu para a síntese de NO. A exposição das mitocôndrias ao NO ou ao nitrito causou uma inibição transiente e menor no consumo de oxigênio quando NAD(P)H f oi utilizado como substrato respiratório, em relação ao succinato. Estas inibições foram potencializadas por superóxido dismutase, sugerindo um mecanismo de degradação de NO dependente de NAD(P)He de oxigênio pelas mitocôndrias de A. thaliana. O conjunto dos resultados mostra que o NO pode ser produzido pela mitocôndria de A. thaliana através de uma atividade redutora de nitrito e que as NAD(P)H desidrogenases e a AOX, além da COX, podem interferir nesta produção. Em aerobiose, a quantidade de NO produzido pode ser controlada pelas atividades de degradação das NAD(P)H desidrogenases e pela COX. A AOX contribui de maneira oposta, ajudando a manter a concentração de NO. Em anaerobiose a atividade redutora de nitrito é favorecida sendo o NO produzido nestas condições pela COX e também devido ao estado mais reduzido da cadeia respiratória. Ainda, em anaerobiose, os mecanismos de degradação de NO, dependentes de oxigênio, estão inibidos. Em conclusão, os resultados deste trabalho mostram a importância das proteínas alternativas da cadeia respiratória da mitocôndria para a homeostase do NO na planta / Abstract: The objective of this work was to study the mechanisms of synthesis and degradation of NO by plant mitochondria, with emphasis on the influence of the NAD(P)H dehydrogenases and the alternative oxidase (AOX) in these processes. The experiments were carried out with mitochondria isolated from cells in culture of the model plant Arabidopsis thaliana. Initially, a protocol for the isolation and purification of mitochondria was established, and their functional integrity was determined by respiratory control, electric membrane potential and electrophoretic calcium transport. The processes of synthesis and degradation of NO were analyzed by monitoring oxygen consumption and NO concentration in the reaction medium with specific electrodes. The results showed that nitrite inhibited respiration of A. thaliana mitochondria, in competition with oxygen in the medium. This inhibition was reversed by cPTIO, an NO scavenger. When AOX was inhibited by n-propylgalate, the inhibitory effect of nitrite in mitochondrial respiration was increased, whereas in the presence of myxothiazol, which prevents the reduction of cytochrome c oxidase (COX), nitrite had no effect on respirati on. The observed effects of nitrite in the oxygen consumption suggested that NO was synthes ized by A. Thaliana mitochondria through a nitrite reductase activity. Meanwhile, the synthesis of NO from nitrite was mainly detected under anaerobiosis. At high concentrations of nitrite (1-5 mM), the production of NO was virtually insensitive to inhibitors of the respiratory chain, while at lower concentrations of nitrite (100-200 ?M) COX, but not AOX, contributed for the synthesis of NO. The exposure of mitochondria to NO or nitrite caused a transient and lower inhibition in oxygen consumption when NAD(P)H was used as respiratory substrate, compared to succinate. These inhibitions were enhanced by superoxide dismutase, suggesting an NAD(P)H- and oxygendependent mechanism for NO degradation by mitochondria of A. thaliana. The overall results show that NO can be produced by A. thaliana mitochondria through a nitrite reductase activity and that NAD(P)H dehydrogenases and AOX, in addition to COX, can interfere in this production. In aerobiosis the amount of NO produced can be controlled by the degradation activities of NAD(P)H dehydrogenases and COX. The AOX has an opposite effect, keeping the concentration of NO. In anaerobiosis the nitrite reductase activity is favore d and NO is produced in these circumstances by COX, and also due the more reduced state of the respiratory chain. Additionally, in anaerobiosis, the oxygen -dependent mechanisms of NO degradation are inhibited. In conclusion, the results of this study show the importance of the alternative proteins of the mitochondrial respiratory chain for NO homeostasis in the plant / Mestrado / Bioquimica / Mestre em Biologia Funcional e Molecular
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Determinação do teor de nitrato em formula infantil por sistema FIA e estimativa da sua exposição pelos lactentesElias, Elede Martins 25 February 2005 (has links)
Orientador : Felix Guillermo Reyes Reyes / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-04T02:35:55Z (GMT). No. of bitstreams: 1
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Previous issue date: 2005 / Mestrado / Ciência de Alimentos / Mestre em Ciência de Alimentos
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Purification, Characterisation And Regulation Of Nitrite Reductase From Candida UtilisSengupta, Sagar 03 1900 (has links) (PDF)
No description available.
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Validação de biomarcadores salivares por meio de parâmetros clínicos e microbiológicos em Periodontia / Validation of salivary biomarkers by clinical and microbiological parameters in PeriodonticsMarcos Vinícius Moreira de Castro 09 April 2014 (has links)
A saliva possui diversas substâncias e propriedades, sendo considerada de importância ao organismo. Possibilita hidratação e lubrificação, fundamentais para integridade da mucosa bucal, atividade remineralizante essencial aos tecidos dentários, apresenta substâncias antimicrobianas dentre outras. Este estudo buscou validar biomarcadores salivares como atividade argininolítica, quantificação de nitritos e proteína total no diagnóstico e após terapia periodontal. Metodologia: No presente estudo do tipo intervencional foram avaliados participantes com gengivite e periodontite crônica na pré (momento inicial) e pós-terapia (noventa dias). Foram efetuados exames clínicos periodontais (profundidade de sondagem, nível clínico de inserção, índice de placa e índice gengival); análise de biomarcadores salivares para valida-los no diagnóstico e após terapia periodontal: atividade argininolítica, quantificação de nitritos e proteína total. Em adição, exames microbiológicos no sulco/bolsa periodontal das espécies do complexo vermelho (Tannerella forsythia, Porphyromonas gingivalis) e Streptococcus oralis por meio de reação em cadeia da polimerase em tempo real. A amostra constou de 57 participantes sistemicamente saudáveis. Sendo 26 diagnosticados como gengivite e tratados por meio de procedimentos básicos aplicando raspagem ultrassônica em sessão única associada com enxaguatório bucal Listerine. Participantes que apresentaram periodontite crônica generalizada foram 31, tratados por procedimentos básicos, incluindo raspagem manual. Destes, 15 realizaram raspagem por quadrante e 16 raspagem em sessão única (full-mouth disinfection), ambos associados com clorexidina a 0,12% como enxaguatório bucal. Resultados: Noventa dias após o tratamento mecânico e químico, todos os grupos apresentaram melhoras clínicas e microbianas. A avaliação da proteína total não apresentou diferença estatisticamente significativa (p<0,05). Não ocorreu redução estatisticamente significativa (p<0,05) de nitrito, enquanto a arginase reduziu estatisticamente (p<0,05) nos grupos gengivite e periodontite full-mouth disinfection. Conclusões: O presente estudo encontrou melhora nos parâmetros clínicos e microbiológicos pós-tratamento mecânico-químico de gengivite e periodontite crônica. Na análise imunológica salivar, a validação de biomarcadores acompanhando esta melhora, ocorreu apenas para atividade argininolítica. / Saliva has several substances and properties, being considered of importance to the organism. Provides hydration and lubrication, fundamentals to the integrity of the oral mucosa, remineralizing activity essential to dental tissues, has among other antimicrobial substances. This study aimed to validate salivary biomarkers as argininolytic activity, quantification of nitrite and total protein in the diagnosis and after periodontal therapy. Methods: In this interventional study participants with gingivitis and chronic periodontitis in pre (initial stage) and after therapy (ninety days) were evaluated. Clinical periodontal examination (probing depth, clinical attachment level, plaque index and gingival index) were performed; analysis of salivary biomarkers to validate them at diagnosis and after periodontal therapy: argininolytic activity, quantification of nitrite and total protein. In addition, microbiological tests in the sulcus/periodontal pocket of the red complex species (Tannerella forsythia, Porphyromonas gingivalis) and Streptococcus oralis by real-time polymerase chain reaction. The sample consisted of 57 systemically healthy study participants. Of these 26 diagnosed as gingivitis and treated through basic procedures applying ultrasonic scaling in a single session associated with Listerine mouthwash. Participants who had chronic periodontitis were 31, treated with basic procedures, including manual scaling. Of these, 15 scaling per quadrant and 16 scaling in only session (full-mouth disinfection), both associated with 0.12% chlorhexidine as mouthwash. Results: Ninety days after the mechanical and chemical treatment, all groups showed clinical and microbial improvement. The assessment of total protein showed no statistically significant difference (p<0.05). There was no statistically significant reduction (p<0.05) of nitrite, while arginase statistically reduced (p<0.05) in gingivitis and periodontitis full-mouth disinfection groups. Conclusion: The present study found improvement in clinical and microbiological parameters in mechanical and chemical post-treatment of gingivitis and chronic periodontitis. In the immunological salivary analysis, the validation of biomarkers watching this improvement occurred only for argininolytic activity.
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Studies of gastric aspirate nitrite, pH, bacterial flora and mutagenicity in manColdrey, Norman A 31 March 2017 (has links)
Gastric aspirate specimens were collected from patients w~th clinically diagnosed gastric carcinoma and from non-carcinoma patients. The nitrite concentration and pH values of the aspirates were measured, the microorganisms present in selected specimens were isolated and identified, and the mutagenicity ratios of the aspirates were determined. The median nitrite concentration of the gastric aspirates from the carcinoma patients was significantly higher than that obtained for the non-carcinoma patients. A positive correlation was found between the nitrite concentration and the pH values of all the specimens tested, and a marked increase in nitrite levels at pH values above 6,0 was evident in specimens from the coloured ethnic "normal" subgroup. Gastric aspirate nitrite concentrations did not correlate with salivary values. The presence of microorganisms in gastric aspirates was shown to be pH dependent. Gastric aspirates with a pH < 2,0 were sterile, below pH 4,0 only acidophilic bacteria survived, whereas above pH 4,0, numerous species, predominantly members of the oral microflora, were isolated. The mean mutagenicity ratio of the gastric aspirates from the carcinoma patients was found to be significantly higher than that found for the control group. There was a positive correlation between the mutagenicity ratios of all the gastric specimens and pH with a maximum at a pH value of approximately 6,0.
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Recherche d'alternatives aux nitrates et nitrites dans les produits carnésKouassi, Anne Patricia 24 April 2018 (has links)
L'objectif principal de cette maîtrise était de développer des formulations d'épices capables de remplacer les nitrites et nitrates dans les produits carnés tout en gardant la même durée de conservation que ces derniers ainsi que leurs propriétés organoleptiques, antibactériennes et antioxydantes. Un criblage qualitatif puis quantitatif des épices a été réalisé et trois épices ont été sélectionnées, les clous de girofle, le cumin et la cannelle. Puis, des poudres de fruits ont été testées pour approcher la couleur rosée que donnent les échantillons contenant les nitrites. L'analyse sensorielle effectuée par la suite a fait ressortir le caractère sucré de la poudre de fruit et des propriétés très intéressantes de certaines formulations par rapport à l'ajout des nitrites. Enfin, une analyse technico-économique a été réalisée et a montré une très faible augmentation du coût de production des produits carnés en utilisant nos formulations d'épices et de poudre de fruit. / The aim of this study was to develop formulations that can replace nitrates and nitrites in meat-based products while keeping the same shelf life as nitrites and also, their organoleptic, antimicrobial and antioxydant properties. A qualitative and quantitative screening of spices have been made and three spices were selected, cloves, cumin and cinnamon. Different fruit powders were tested to get closer to the pinkish color which nitrites give to meats. Then, an organoleptic tests were performed on a panel of tasters and the results have helped identify the sweetness of the variety of fruit powder and very interesting sensory properties of several formulations in relation to the addition of nitrites. Finally, a techno-economic analysis was performed and showed a small increase in cost of production of meat products using spices and fruit powder, due to a slight increase in the cost of raw materials.
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Šachtinių šulinių geriamojo vandens, naudojamo nėsčių moterų ir kūdikių, kokybė ir jogerinimo galimybės Pakruojo rajone / Dug wells for drinking water used by pregnant women and infants, the quality of the water and opportunities for improvement in pakruojis districtŽuvininkienė, Vilma 21 June 2010 (has links)
Darbo tikslas – įvertinti šachtinių šulinių geriamojo vandens kokybę ir jo gerinimo galimybes Pakruojo rajone.
Uždaviniai:
1. Įvertinti šachtinių šulinių geriamojo vandens kokybės rodiklius.
2. Nustatyti nėščių moterų ir moterų, kurios augina kūdikius iki 6 mėn. amžiaus, požiūrį į naudojamo vandens kokybę ir tyrimo rodiklius.
3. Įvertinti šachtinių šulinių geriamojo vandens kokybės gerinimo galimybes.
Tyrimo metodika. Anketinė nėščių moterų ir kūdikius iki 6 mėn. amžiaus auginančių moterų (n=167, atsako dažnis 100 proc.) apklausa. Šachtinių šulinių geriamojo vandens kokybės rodiklių tyrimo protokolų analizė (n = 270) ir visuomenės sveikatos centro ir biuro specialistų, rajono ekologo (n=3, atsako dažnis 100 proc.) interviu. Apklausos duomenys analizuoti „SPSS 15.0 for Windows“ programa. Hipotezės tikrintos, taikant χ² testą, nustatčius reikšmingumo lygį p<0,05.
Rezultatai. Tyrimo metu buvo nustatyta, kad respondentės, naudojančios užterštų šachtinių šulinių vandenį, dažniausiai gyvena kaimo vietovėse, turi mažas metines pajamas, yra bedarbės. 95,8 % respondenčių gavo informaciją apie šachtinių šulinių vandens kokybę ir patarimus, kaip apsaugoti save ir kūdikius. Moterys, turinčios žemesnį išsilavinimą, informavus apie vandens tyrimo rezultatus bei riziką sveikatai, vartoja užterštą geriamąjį vandenį nitratais ir nesiima jokių veiksmų jo kokybei gerinti. Netinkamai tvarkoma šulinio aplinka, kuri gali sąlygoti užteršimą: apie šulinį auginami augalai, kurie yra tręšiami... [toliau žr. visą tekstą] / The Aim of the study: to asses the quality of the possibilities of improving it in Pakruojis district.
Objectives :
1. Estimate the quality of the dug wells for drinking water.
2. Evaluate the standpoint of pregnant women and the women growing up babies sunder the age of six (6) months to the dug wells water quality indicators.
3. Estimate the opportunities of improvements for the quality of drinking water.
Research methods. Questions for pregnant women and the women raising infants under six months Cn =167, response rate was 100 percent survey. The analysis of the dug wells for drinking water quality indicators (n=270) made by the public health center, the office professionals the region ecologist (n=3, response rate 100 percent) interview. Survey data, was analysed at “SPSS15.0 for Windows” program. The Hypothesis is verified by using the test χ2. According to it the significance level is p<0,05.
Results. The study found out that the respondents using contaminated water from dug wells, are unemployed. 95.8 % of them have received the right information about the quality of the water in their dug wells. They also have get the advices how to protect themselves and their children. The women with lower education were informed about the test results of their water, about health risks using the water, they still consumpt contaminated by nitrates drinking water and don’t take any actions to improve its quality. Improperly handled wells, their environment which may lead to... [to full text]
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The effects on the performance of broilers consuming calcium, potassium, and sodium nitrates and nitrites from the drinking waterReeder, Julie A. 10 May 1996 (has links)
Four experiments were carried out with broiler chicks from day-old to three or
four weeks of age to assess the effects of nitrates and nitrites from calcium,
potassium, and sodium salts in the drinking water of broilers on growth, body
weights, feed utilization, blood chemistries, liver tissues, and lipid oxidation of breast
and thigh tissues. Body weights were lower (P<.05) among chicks consuming
calcium nitrate than those of chicks consuming sodium nitrate or the control. Chicks
consuming calcium nitrate (Ca(NO���)���) at 1000 ppm had lower (P<.05) body weights
than those drinking Ca(NO���)��� at 0, 50, or 200 ppm. Broilers receiving 1000 ppm of
calcium nitrite (Ca(NO���)���) had depressed body weights when compared to chicks
consuming 0, 50, or 200 ppm of Ca(NO���)���. Consumption of sodium nitrate (NaNO���)
at 2033 ppm reduced (P<.05) broiler weights in comparison to broilers ingesting 0,
111, or 427 ppm of NaNO���. Feed utilization was less efficient (P<.05) by chicks
ingesting Ca(NO���)��� and calcium, potassium, or sodium salts of nitrite when compared
with the control. / Graduation date: 1996
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Estudo experimental das alterações morfológicas do intestino grosso em ratos submetidos à operação de derivação jejunoileal / Experimental study of the morphological changes of the large intestine in rats submitted to jejunoileal bypass operationBraoios, Maria Cristina Costa Resck 02 December 2010 (has links)
Orientador: Nelson Adami Andreollo / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-15T23:47:26Z (GMT). No. of bitstreams: 1
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Previous issue date: 2010 / Resumo: As gastroplastias e as derivações intestinais revolucionaram o tratamento da obesidade mórbida, pela sua viabilidade e resposta sustentada. Porém, estudos experimentais, sugerem após as derivações jejunoileais um aumento do risco de câncer de cólon devido ao aumento do tamanho e da proliferação das criptas intestinais. Evidências sugerem os possíveis efeitos antioxidantes na prevenção da carcinogênese que demonstram sua capacidade de neutralizar ou eliminar os radicais livres. O ácido ascórbico participa do processo de oxirredução das células, como também é importante na biossíntese de catecolaminas. Os compostos de nitrito e nitrato são sais empregados como conservantes de alimentos, que neste contexto do aumento de população mundial e escassez de alimentos, será cada vez mais empregado. Estes sais têm ação danosa ao organismo em situações específicas, como demonstram vários experimentos. O objetivo dessa pesquisa foi analisar as alterações histopatológicas caracterizadas pelo índice de mitoses produzidas no intestino grosso de ratos Wistar submetidos à derivação jejunoileal após administração continuada de vitamina C e nitrito de sódio, em diferentes grupos. Oitenta ratos machos Wistar pesando entre 450-550g foram divididos em doze grupos, com vinte animais controles não operados (5 animais ingeriram somente água, 5 animais ingeriram somente vitamina C, 5 animais ingeriram somente nitrito de sódio e 5 animais ingeriram vitamina C+nitrito de sódio). Sessenta animais foram anestesiados e submetidos à laparotomia mediana com exposição das alças intestinais. Em vinte animais apenas manipulação intestinal foi realizada, este servindo como sham. Nos restantes (grupo denominado operado), o intestino foi seccionado a 3cm da junção duodeno-jejunal, e a derivação jejunoileal foi realizada através da anastomose látero-lateral no íleo terminal a 2cm da válvula íleo-cecal. Os animais foram acondicionados em gaiolas com cinco animais cada de acordo com a droga que receberam. Após 180 dias de observação, os animais eram pesados semanalmente, e para análise foram divididos em segmentos (S1, S2, S3, S4 e S5) correspondentes ao ceco, colon ascendente, transverso, descendente e reto. No grupo controle, observando os segmentos em comparação ao tratamento (p< .0001) e comparando a média entre todos os segmentos (p=0,0323) indica que o nitrito+vitamina C obtiveram o menor índice de mitoses em relação aos outros tratamentos. No grupo operado os segmentos S1 e S2 houve diferença estatística com o tratamento vitamina C, o qual apresentou um maior índice de mitose e melhor preservação do epitélio. No grupo sham houve diferença apenas entre tratamento (p< .0001), não havendo diferença entre segmentos (p=0,0849) e nem interação entre segmento e tratamento (p=0.7135). A principal diferença estatística ficou apenas no grupo nitrito+vitamina C entre a média dos segmentos. Concluem que, comparando todos os segmentos colônicos avaliados dos vários grupos estudados, foi observado que os animais que receberam nitrito de sódio associado à vitamina C apresentaram menor índice de mitoses. Além disso, foi verificado que a vitamina C não apresentou um efeito inibidor, estatisticamente significativo, na preservação da mucosa e no índice de mitos / Abstract: Gastroplasty and intestinal bypass revolutionized the treatment of morbid obesity, the viability and sustained response. However, experimental studies suggest following the jejunoileal bypass increased risk of colon cancer due to the increased size and proliferation of intestinal crypts. Evidence suggests the possible effects of antioxidants in the prevention of carcinogenesis that demonstrate their ability to neutralize or eliminate free radicals. The ascorbic acid redox process of the cells, it is also important in the biosynthesis of catecholamines. The compounds of nitrite and nitrate salts are used as food preservatives, which in this context of increasing world population and food shortages, will be increasingly employed. These salts have a harmful action to the body in specific situations, as demonstrated by several experiments. The aim of this study was to evaluate the histopathological changes characterized by the index of mitosis produced in the intestine of rats subjected to jejunum-ileum bypass after continuous administration of vitamin C and sodium nitrite in different groups. Eighty male Wistar rats weighing 450-550g were divided into twelve groups, with twenty non-operated control animals (5 animals ingested only water, only 5 animals ate vitamin C, only 5 animals ingested sodium nitrite and 5 animals ate vitamin C + nitrite sodium). Sixty animals were anesthetized and underwent laparotomy with exposure of the bowel. In twenty animals only intestinal manipulation was performed, this serving as a sham. In the other (called the operated group), the intestine was cut to 3 cm from the duodenum-jejunal junction, and jejunum-ileum bypass was performed through the side-to-side anastomosis in the ileum 2 cm from the ileocecal valve. The animals were placed in cages with five animals each according to the drug they received. After 180 days of observation, the animals were weighed weekly, and for analysis were divided into segments (S1, S2, S3, S4 and S5) corresponding to the cecum, ascending colon, transverse, descending and rectum. In the control group, noting the segments in comparison to treatment (p <.0001) and comparing the average of all segments (p = 0.0323) indicates that nitrite + vitamin C obtained the lowest rate of mitosis in relation to other treatments. In the group operated the segments S1 and S2 was no statistical difference in treatment vitamin C, which showed a higher rate of mitosis and better preservation of the epithelium. In the sham group only showed a difference between treatment (p <.0001), with no difference between segments (p = 0.0849) nor interaction between segment and treatment (p = 0.7135). The main difference was statistically only for the nitrite + vitamin C between the average of the segments. They conclude that, comparing all colonic segments evaluated the various groups, it was observed that the animals that received sodium nitrite plus vitamin C had a lower rate of mitosis. Furthermore, it was found that vitamin C did not show an inhibitory effect was statistically significant to the preservation of the mucosa and the index of mitos / Doutorado / Pesquisa Experimental / Doutor em Cirurgia
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Capillary Ion Electrophoresis of Endogenous Anions and Anionic Adulterants in Human UrineFerslew, K. E., Hagardorn, A. N., Robert, T. A. 01 January 2001 (has links)
Normal human urine contains many anions and cations. Ionic concentrations in urine have classically been determined by spectrophotometry of color reactions, flame emission spectrophotometry, atomic absorption spectrophotometry, high performance liquid chromatography, or potentiometry with ion-specific electrodes. Capillary ion electrophoresis (CIE) is a form of capillary electrophoresis which uses the differential electrophoretic mobility of ions to perform a separation of an ionic mixture. Various salts can be added to urine specimens to abnormally elevate ionic concentrations and interfere with either immunoassay urine drug screening procedures or gas chromatographic/mass spectrometric confirmation techniques. Application of CIE for the direct detection of endogenous anions and anionic adulterants in human urine specimens was the purpose of this investigation. CIE was performed using a Waters Quanta 4000 Capillary Electrophoresis System with either direct or indirect ultraviolet absorption detection at 254 nm. CIE of 30 random normal urine specimens and 21 urine specimens suspected of adulteration was performed. Duplicate aliquots were assayed by CIE and by colorimetric technique for nitrite. Sixteen specimens had elevated concentrations of nitrite and/or nitrate. The correlation coefficient between nitrite CIE and colorimetric results was 0.9895. Three specimens had detectable concentrations of chromate and were suspected of being adulterated with "Urine Luck," an adulterant found to contain chromate. Two specimens suspected of being adulterated with bleach were found to only contain chloride, sulfate, and phosphate. CIE is applicable to forensic analysis of urine anion concentrations. CIE can easily quantitate numerous endogenous anions and offers a method to detect and/or confirm anion adulteration of urine specimens.
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