Thesis (Ph. D.)--University of Wisconsin--Madison, 1966. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references.

Aristolochic acids (AA) have been considered as toxicants of plants which were found in plants of the family Aristolochiaceae. The most abundant acids in mentioned plants are aristolochic acid I (AAI) and aristolochic acid II (AAII). AA have been considered as causes kidney disease called Aristolochic acid nephropathy (AAN). AAN was initially discovered in patients of one Belgian clinic in Brussels specialized on treatment of patients leading to a decrease in their body weight. The first name of this disease was Chinese herb nephropathy (CHN). Later, it was discovered that one component of herbal preparation was changed by a mistake with the Aristolochiaceae plant. The second type of renal disease caused by AA was discovered in populations of countries along the Danube river, called as Balkan endemic nephropathy (BEN), which was probably caused by the contamination of grains with plants containing AA. These renal diseases (AAN and BEN) are often associated with development of upper urothelial cancer (UUC). AA (AAI + AAII) in organisms are subject to biotransformation leading to its reductive activation or oxidative detoxification. Both cytosolic enzymes [NAD(P)H:quinone oxidoreductase] and microsomal enzymes [cytochromes P450, NADPH:cytochrome P450 reductase] participate in their reduction. The...

Utilisation de la métabonomique en spectroscopie RMN pour l'identification de biomarqueurs d'exposition à l'acide Aristolochique. Développement de modèles expérimentaux chez des rats mâles. Analyse d'échantillons urinaires provenant de patients croates potentiellement touchés par la Néphropathie endémique des Balkans / Doctorat en Sciences biomédicales et pharmaceutiques (Médecine) / info:eu-repo/semantics/nonPublished

Aggregative feeding is widespread in Lepidopteran larvae suggesting that this behavior serves on adaptive function. Many studies of the potential benefits of aggregative feeding in Lepidopteran larvae have been conducted. However, no studies have directly examined the benefits of cryptic larvae being both chemically defended and gregarious. Group feeding occurs disproportionately more in chemically defended larvae than in larvae that have no chemical defense. Most of these larvae are cryptic when they are most highly aggregated and most vulnerable to predation. In this study, the benefits of group feeding in terms of decreased predation were explored in first instar larvae of pipevine swallowtail larvae, Battus philenor, a species that exhibits chemical sequestration. Contrary to our expectation, we found that groups of larvae fed a diet with high levels of the toxin aristolochic acid, which they sequester naturally and use as a defense against natural enemies, had significantly lower survivorship due to predation in both the field and in the laboratory experiments compared to groups of larvae fed a diet with low aristolochic acid content. We also found that aristolochic acid does not deter the generalist predator Hippodamia convergens, the ladybird beetle, suggesting that this compound is not a universal predator deterrent as previously assumed. Thus, instead of finding a benefit to group feeding and chemical defense in cryptic larvae, we have found a negative impact of group feeding in this population of B. philenor. Based on this evidence, we speculate that other benefits of group feeding might be outweighing the negative consequences of increased predation during the first instar. Future research on chemical defense, aposematism, and aggregative feeding should take into consideration that chemical defenses might not be universally effective against all natural enemies.

Aristolochic acids (AA) are human carcinogens which have also very strong nephrotoxic properties. A mixture of AA is present in Aristolochiacae plant species. These plants were and still are used in traditional medicine in some countries, particularly in Asia. Aristolochic acids participate in development of two types of nephropathies. The first disease is designated as Aristolochic Acid Nephropathy (AAN), the second one is Balkan Endemic Nephropathy (BEN). Both nephropathies are associated with urothelial malignancies, which are caused by AA. One of the common features of ANN and BEN is that not all individuals exposed to AA suffer from nephropathy and tumour development. One cause for these different responses may be individual differences in the activities and expression levels of the enzymes catalyzing the biotransformation of AAI, the major toxic component of AA contained in Aristolochia species. Detailed knowledge of enzymes which participate in metabolism of AAI may contribute to elucidation of inter-individual susceptibility to AAN, BEN and later urothelial malignancies. Aristolochic acid I is either oxidative detoxicated or reductive activated by biotransformation enzymes. Reductive bioactiovation of AAI leads to formation of covalent AA-DNA adducts in organism which result in producing of...

ABSTRACT Cytochrome b5 (CYB5) is heme protein capable of reduction of cytochromes P450 (CYP) or some other enzymes. However, his regulative capability was also observed by his apo form, i.e. in absence of heme prosthetic group in the active center. CYB5 can accept electron from cytochrome b5 reductase (CYB5R) or from cytochrome P450 reductase (CYPOR). CYPOR by itself is reduced by NADPH and is also able to forward electron to CYP independently of CYB5. CYB5R on the other hand is reduced by NADH. Efficiency of CYB5 to accept and forward an electron was studied in vitro with five different substrates - testosterone, Sudan I, aristolochic acid I (AAI), ellipticine and vandetanib. These substrates were chosen considering their characteristic reactions, which are catalyzed by their respective isoforms of CYP. The experiments with these substrates were carried out in the medium with recombinant CYPs prepared in insect cells or E. coli or in the medium with hepatic microsomes isolated from different organisms. Rats, from which the majority of these microsomes was isolated, were premedicated by different CYP inducers. The experiments were carried out in medium with NADH or NADPH in order to assess the capability of CYB5 to reduce CYP independently of CYPOR. The capability of CYB5 and CYB5R to act as a...

La néphropathie aux plantes chinoises (CHN) est une maladie rénale grave qui a été décrite pour la première fois en 1993 chez des patientes ayant suivi un régime amaigrissant à base d’extraits de plantes chinoises (Aristolochia fangchi) contenant des acides aristolochiques (AA). Cette néphropathie se caractérise par une atrophie tubulaire et une fibrose interstitielle aboutissant à l’urémie terminale et se complique fréquemment de cancers des voies urinaires. Au moment d’initier ce travail, il subsistait toujours un large débat quant au rôle étiologique réel des acides aristolochiques dans la genèse de cette maladie. En effet, les gélules à visée amaigrissante contenaient d’autres substances potentiellement néphrotoxiques. Mais surtout, il n’existait aucune preuve expérimentale que les AA pouvaient induire une fibrose rénale interstitielle. Dans la première partie de ce travail, nous démontrons que l’injection par voie sous-cutanée d’AA à la dose de 10 mg/Kg/jour à des rats Wistar mâles en déplétion sodée entraîne l’apparition au 35ème jour d’une atrophie tubulaire, d’une fibrose interstitielle et d’une insuffisance rénale, reproduisant ainsi les anomalies caractéristiques de la CHN. Nous avons ensuite montré que la dexfenfluramine, substance anorexigène à action de type sérotoninergique prise concomitamment par les patientes atteintes de CHN, ne potentialise pas la toxicité rénale des AA. Enfin, la stimulation du système rénine angiotensine (SRA) par la déplétion sodée ou l’inhibition de celui-ci par un traitement pharmacologique ne modifie pas la fibrose interstitielle ni l’insuffisance rénale induite par les AA. En conclusion, nous avons réussi à développer un modèle in vivo de fibrose rénale interstitielle induite par les AA. Dès lors nous avons apporté la preuve expérimentale de l’implication des AA dans le développement de la CHN. Ce modèle a permis de démontrer que les autres éléments potentiellement néphrotoxiques contenues dans la cure d’amaigrissement (dexfenfluramine, diurétique, laxatif) n’influençaient pas l’évolution de la fibrose interstitielle, ce qui confirme que la prise isolée d’AA suffit à expliquer le développement de la CHN. Cette confirmation à d’importantes implications en santé publique dans la mesure où des plantes contenant des acides aristolochiques font toujours partie des phytothérapies traditionnelles. De plus, il est apparu que, dans ce modèle, les mécanismes de la fibrose rénale interstitielle pouvaient être largement indépendants du SRA. Enfin, de par sa durée limitée et sa grande reproductibilité, ce modèle constitue un outil expérimental d’avenir pour l’étude des mécanismes physiopathologiques de la fibrose rénale interstitielle en général.

Aristolochic acids (AA) are carcinogenic and nephrotoxic alkaloids from Aristolochia species. Aristolochic acid I (AAI), the major component of AA, causes the development of Aristolochic acid nephropathy (AAN) and Balkan endemic nephropathy (BEN). These two diseases cause total renal failure and urothelial malignancies. The fact that these diseases have not been developed in all persons, who have been exposed to their action, might be causd by different activities and protein levels of the enzymes metabolizing AAI. Thus, the identification of enzymes involved in the metabolism, and detailed knowledge of their expression and catalytic specifities is a major importance. Aristolochic acid I (AAI) can be metabolized by several types of reactions. Like most nitroaromatics, the main activation pathway of AAI is reduction of its nitro group to form a cyclic acylnitrenium ion, which can bind to the purine bases, thereby forming AAI-DNA adducts. The detoxication pathway of AAI is its oxidative demethylation by cytochromes P450 forming detoxication metabolite 8-hydroxyaristolochic acid Ia (AAIa). In the present thesis, using rat and human enzymes and as well as several mice models, the metabolism of AAI in vitro and in vivo was investigated. The first model has deleted gene for NADPH:cytochrome P450...

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5.4pt;mso-para-margin-top:0in;mso-para-margin-right:0in;mso-para-margin-bottom:10.0pt;mso-para-margin-left:0in;line-height:115%;mso-pagination:widow-orphan;font-size:11.0pt;font-family:"Calibri","sans-serif";mso-ascii-font-family:Calibri;mso-ascii-theme-font:minor-latin;mso-fareast-font-family:"Times New Roman";mso-fareast-theme-font:minor-fareast;mso-hansi-font-family:Calibri;mso-hansi-theme-font:minor-latin;}</style><![endif]--></p><p class="MsoNormal" style="text-align:justify">Uvod: Aristolohična kiselina I je nefrotoksična i kancerogena supstanca koja je odgovorna za nefropatiju koja nastaje usled kori&scaron;ćenja herbalnih preparata i čajeva za mr&scaron;avljenje. S obzirom da se ova supstanca može naći u korovskim biljkama, smatra se jednim od glavnih ekotoksikolo&scaron;kih uzroka za nastanak balkanske endemske nefropatije čiji definitivan uzrok jo&scaron; uvek nije otkriven. Toksičnost ove supstance je dokazana na brojnim animalnim modelima, međutim mehanizmi koji dovode do o&scaron;tećenja bubrežnog parenhima jo&scaron; u potpunosti nisu razja&scaron;njeni.<span style="mso-spacerun:yes">&nbsp; </span>Cilj: Doktorska disertacija je koncipirana sa ciljem da se utvrdi uticaj toksičnog jedinjenja aristolohične kiseline I na histopatolo&scaron;ke i imunohistohemijske karakteristike tubulointersticijuma i glomerula bubrega kod mi&scaron;eva, kao i na biohemijske parametre krvi i urina koji ukazuju na o&scaron;tećenje bubrega. Materijal i metode: U ekperimentu je kori&scaron;ćeno 64 mi&scaron;a soja NMRI koji su podeljeni u tri grupe: eksperimentalna grupa (n=32) koja je dobijala aristolohičnu kiselinu I rastvorenu u polietilen glikolu (2,5% PEG 400) u dozi od 10 mg/kg telesne mase, negativna kontrolna grupa koja je dobijala 2,5% PEG 400 (n=16) i kontrolna grupa koja je dobijala fiziolo&scaron;ki rastovor (n=16). Sve životinje su tretirane intraperitonealno svakodnevno tokom sedam dana. Tokom eksperimenta 8., 17., 29. i 59. dana sakupljan je dvadesetčetvoročasovni urin 8 životinja iz eksperimentalne grupe, 4 životinje iz negativne kontrolne i 4 životinje iz kontrolne grupe. Životinje su žrtvovane 9., 18., 30. i 60. dana, uzeta im je krv, dok su bubrezi posebno odvojeni radi histopatolo&scaron;ke analize. Na bubrežnom tkivu sprovedene su histohemijske, imunohistohemijske i morfometrijske analize, dok su na uzorcima seruma i urina sprovedene biohemijske analize. Dobijeni rezultati su testirani adekvatnim statističkim metodama i prikazani su tabelarno i grafički. Rezultati: Nefrotoksin aristolohična kiselina I nakon 7 dana aplikacije izaziva značajno o&scaron;tećenje bubrežnog parenhima. Pri aplikaciji 2,5% PEG 400 i fiziolo&scaron;kog rastvora ne dolazi do vidljivog o&scaron;tećenja bubrežnog parenhima. Histopatolo&scaron;ku sliku u ranoj fazi eksperimenta (9. i 18. dan) karakteri&scaron;e akutna tubulska nekroza proksimalnih tubula. U kasnijoj fazi (30. i 60. dana) uočava se histopatolo&scaron;ka slika hroničnog intersticijalnog nefritisa sa obilnim mononuklearnim ćelijskim infiltratima limfocitnog porekla kao i postojanje blage intersticijalne fibroze. Kod eksperimentalnih životinja je morfometrijskim metodama utvrđen veći stepen bubrežnog o&scaron;tećenja tubulointersticijuma i smanjen broj podocita u glomerulu u odnosu na kontrolne grupe. Biohemijske analize kod većine eksperimentalnih životinja su pokazale veće koncentracije serumske uree nego kod kontrolnih grupa. Takođe je dokazana albuminurija u kasnijoj fazi eksperimenta koja je veća kod životinja izloženih aristolohičnoj kiselini I nego kod životinja iz kontrolnih grupa. Zaključak: Kori&scaron;ćenjem morfometrijskih metoda u okviru histopatolo&scaron;kih i imunohistohemijskih ispitivanja, uz adekvatne biohemijske analize, može se zaključiti da je aristolohična kiselina I izuzetno nefrotoksično jedinjenje koje izaziva izrazite<span style="mso-spacerun:yes">&nbsp; </span>promene tubulointersticijuma i glomerula. Podaci ovog istraživanja predstavljaju polaznu osnovu za dalja istraživanja dijagnostike u ranoj fazi nefropatija izazvanih aristolohičnim kiselinama.<span style="mso-spacerun:yes">&nbsp; </span></p> / <p>Introduction: Aristolochic acid I is a nephrotoxic and carcinogenic substance responsible for nephropathy caused by the use of herbal preparations and teas for slimminng regimen. Since this substance can be found in plants, it is considered one of the major ecotoxicological causes for the emergence of balkan endemic nephropathy whose definitive cause has not yet been revealed. The toxicity of this substance has been proven on numerous animal models, but pathophysiological mechanisms of kidney injury still remain unclear. Aim: The doctoral dissertation was designed to determine the influence of aristolochic acid on the histopathological and immunohistochemical characteristics of tubulointerstitium and glomerulus in mice, as well as the biochemical parameters of blood and urine that indicate kidney injury. Material and methods: For this study, 64 mouse of NMRI strain is used. They are divided into three groups: an experimental group (n=32) that received aristolochic acid I dissolved in polyethylene glycol (2.5% PEG 400) at a dose of 10 mg/kg of body weight, a negative control group that received 2.5% PEG 400 (n=16) and a control group that received only saline (n=16). All animals were treated intraperitoneally daily for seven days. During the experiment on the 8th, 17th, 29th and 59th day, twenty-four-hour urine was collected from 8 animals from the experimental group, 4 animals from the negative control and 4 animals from the control group. Animals were sacrificed on the 9th, 18th, 30th and 60th days, their blood was taken, while the kidneys were taken for histopathological analysis. Histochemical, immunohistochemical and morphometric analyzes were performed on renal tissue, while biochemical analyzes were performed on serum and urine samples. Obtained results were tested with adequate statistical methods and presented in a tables and graphs. Results: After 7 days of application nefrotoxin aristolochic acid I causes significant kidney injury. After application of 2.5% PEG 400 and saline, there was no visible damage to kidney parenchyma. Histopathological changes at the early stage of the experiment (9th and 18th day) were characterized by acute tubular necrosis of proximal tubules. At a later stage (30th and 60th day), chronic interstitial nephritis was observed in kidneys, with abundant mononuclear cell infiltrates in interstitium and presence of mild interstitial fibrosis. In experimental animals, a higher tubulointerstitial score of kidney injury and a decrease in the number of the podocytes in glomerulus were determined by morphometric methods, compared to the control groups. Biochemical analyzes in most experimental animals showed higher blood urea nitrogen concentrations than in control groups. High concentration of albumin in urine can be found in later stages of the experiment, and those concentrations were higher in animals exposed to aristolochic acid I than in animals from control groups.&nbsp; Conclusion: Using morphometric, histopathological and immunohistochemical methods, with adequate biochemical analysis, aristolochic acid I is proven to be an extremely nephrotoxic compound that causes drastic changes in tubulointerstitium and glomeruli of kidney parenhyma. Data from this study can be used for further research into early diagnosis of aristolochic acid nephropathy.</p>

Les cellules cancéreuses sont caractérisées par des altérations de l'ADN causées par des facteurs exogènes, comme l'exposition à des agents environnementaux tels que le tabac ou les UV, ou par des mécanismes endogènes tels que les erreurs de polymérase lors de la réplication de l'ADN. L'analyse des causes et des conséquences de ces altérations permet de mieux comprendre les facteurs et mécanismes à l'origine du développement d'un cancer. Les technologies de séquençages à haut débit offrent l'opportunité d'étudier la nature précise de ces altérations à l'échelle du génome et permettent de révéler des signatures mutationnelles distinctes et spécifiques de cancérigènes, fournissant ainsi des hypothèses sur l'étiologie des cancers.L'objectif de ma thèse a consisté à développer des méthodes et des outils bioinformatiques accessibles et conviviaux permettant de faciliter l'analyse et l'interprétation des signatures mutationnelles à partir de données de séquençage à haut débit. L'application de ces outils et méthodes à des séries originales de tumeurs humaines et de systèmes expérimentaux de mutagénèse et carcinogénèse a permis de mieux caractériser la signature mutationnelle de l'acide aristolochique (AA) ainsi que d'autres cancérigènes d'intérêt / Cellular genomes accumulate alterations following exposures to exogenous factors, like environmental agents such as tobacco smoking or UV, or to endogenous mechanisms such as DNA replication errors. Analysing the causes and consequences of these changes allows a better understanding of the mechanisms underlying cancer development and progression. Next-generation sequencing (NGS) technologies provide the opportunity tostudy the nature of the resulting alterations on a genome-wide scale and started to reveal distinct mutational signatures specific to past carcinogenic exposures providing clues on cancer aetiology.The aim of my thesis was to develop user-friendly bioinformatic tools and methods for facilitating the analysis and interpretation of carcinogen-specific mutational signatures from NGS data. Applying these tools and methods to human tumours and experimental models of mutagenesis led to a better characterisation the mutational signature of aristolochic acid (AA), as well as other carcinogens of interest