Construção de modelos de interação in silico e in vitro do inibidor do tipo Kunitz de Adenanthera pavonina L. para as enzimas cisteínicas e serínicas

Serines proteinases inhibitors (PIs) are widely distributed in nature and are able to inhibit both in vitro and in vivo enzymatic activites. Seed PIs in than leguminous are classified in seven families, Bowman-Birk and Kunitz type families that most studied representing an important role in the first line of defense toward insects pests. Some Kunitz type inhibitors possess activities serine and cysteine for proteinases named bifunctional inhibitor, as ApTKI the inhibitor isolate from seed of Adenanthera pavonina. The A. pavonina inhibitor presenting the uncommon property and was used for interaction studies between proteinases serine (trypsin) and cysteine (papain). In order to determinate the in vitro interaction of ApTKI against enzymes inhibitor purification was carried cut by using chromatographic techniques and inhibition assays. The 3D model of the bifunctional inhibitor ApTKI was constructed SWISS-MODEL program by homology modeling using soybean trypsin inhibitor (STI, pdb:1ba7), as template which presented 40% of identity to A. pavonina inhibitor. Model quality was evaluated by PROCHECK program. Moreover in silico analyzes of formed complex between the enzymes and ApTKI was evaluated by HEX 4.5 program. In vitro results confirmed the inhibitory assays, where the inhibitor presented the ability to simultaneously inhibit trypsin and papain. The residues encountered in the inhibitor model of folder structural three-dimensional that make contact to enzymes target coud explain the specificity pattern against serine and cysteine proteinases

Renal- and calcium-dependent vascular effects of Polybia paulista wasp venom

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Effects of Compounds from Passiflora edulis Sims f. flavicarpa Juice on Blood Coagulation and on Proteolytic Enzymes

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Progression of nephropathy after islet of langerhans transplantation in alloxan-induced diabetic rats

We studied the effects of islet of Langerhans transplantation (IT) on the kidney lesions of rats with alloxan-induced diabetes. Forty-five inbred male Lewis rats were randomly assigned to 3 experimental groups: group Gl included 15 non-diabetic control rats (NC), group GIT included 15 alloxan-induced diabetic rats (DC), and group III included 15 alloxan-induced diabetic rats that received pancreatic islet transplantation prepared by nonenzymatic method from normal donor Lewis rats and injected into the portal vein (IT). Each group was further divided into 3 subgroups of 5 rats which were sacrificed at 1, 3, and 6 months of follow-up, respectively. Clinical and laboratorial parameters were recorded in the mentioned periods in the 3 experimental groups. For histology, the kidneys of all rats of each subgroup were studied and 50 glomeruli and 50 tubules of each kidney were analyzed using light microscopy by two different investigators in a double blind study. The results showed progressive glomerular basement membrane thickening (GBMT), mesangial enlargement (ME), and Bowman's capsule thickening (BCT) in the 3 experimental groups throughout the follow-up. These alterations were significantly more severe in DC rats at 6 months when compared to NC rats (p < 0.01). However, the degree of GBMT, ME, and BCT observed in DC rats was not statistically different from IT rats at 1, 3, and 6 months. In addition, Armanni-Ebstein lesions of the tubules (AE) and tubular lumen protein (PRO) observed in DC rats were also observed in IT rats all over the study. These lesions were never present in NC rats. We conclude that IT did not prevent progression of kidney lesions in alloxan-induced diabetic rats within 6 months after transplantation.

Effect of pancreas transplantation on the prevention of nephropathy in alloxan-induced diabetic rats

We studied the effects of pancreas transplantation on kidney lesions of rats with alloxan-induced diabetes. Ninety inbred male Lewis rats were randomly assigned to 3 experimental groups: group NC included 30 non-diabetic control rats, group DC included 30 alloxan-induced diabetic control rats, and group PT included 30 alloxan-induced diabetic rats that received pancreas transplants from normal donor Lewis rats. Each group was further divided into 3 subgroups of 10 rats which were sacrificed at 1, 3, and 6 months of follow-up, respectively. Clinical and laboratory parameters during these periods were documented. The kidneys of 5 rats in each subgroup were studied and 50 glomeruli and tubules from each kidney were analyzed by light microscopy by two different investigators in a double-blind study. There was progressive glomerular basement membrane thickening (GBMT), mesangial enlargement (ME), and Bowman's capsule thickening (BCT) in kidneys of rats in the 3 experimental groups during follow-up. These alterations were significantly higher in DC rats (GBMT: 1.99 +/- 0.31; ME: 2.00 +/- 0.33; BCT: 1.88 +/- 0.27) when compared to NC(GBMT: 1.54 +/- 0.30; ME: 1.56 +/- 0.47; BCT: 1.36 +/- 0.35) and PT rats (GBMT: 1.49 +/- 0.29; ME: 1.57 +/- 0.36; BCT: 1.35 +/- 0.28) at 6 months (P<0.01). The extent of GBMT, ME, and BCT observed in DC rats at 1 and 3 months was not significantly different from NC and PT rats. The amount of kidney lesions in PT rats was similar to that of NC rats and lower than those of DC rats at 6 months (P<0.01). In addition, Armanni-Ebstein lesions of the tubules (AE) and tubular lumen protein (PRO) observed in DC rats were not present in NC or PT rats. We conclude that pancreas transplantation in alloxan-induced diabetic rats prevents the development of kidney lesions beginning at 6 months after transplantation.

Anemia hemolítica causada por Indigofera suffruticosa (Leg. Papilionoideae) em bovinos

As partes aéreas de Indigofera suffruticosa Mill. (família Leg. Papilionoideae), planta incriminada pelos criadores de diversas áreas do Nordeste por doença caracterizada por hemoglobinúria em bovinos, foram administradas por via oral a seis bovinos, em doses diárias repetidas de 10 a 40 g/kg, Todos os animais experimentais apresentaram hemoglobinúria, porém passageira, apesar continuidade da administração da planta. Dois desses bovinos não apresentaram manifestações adicionais, um terceiro animal evidenciou manifestações leves, e os três outros, sintomas adicionais de intensidade moderada: apatia, mucosas visíveis de coloração esbranquiçada, pêlos arrepiados, anorexia, diminuição da freqüência e intensidade dos movimentos ruminais, taquicardia, pulso venoso positivo e dispnéia. Antes da crise hemolítica a urina apresentava coloração verde azulada. Nenhum animal experimental morreu, porém um foi sacrificado durante a fase hemoglobinúrica. À necropsia observaram-se anemia, bexiga contendo urina cor de vinho tinto, rins aumentados de volume com coloração marrom-escura, fígado, na superfície e ao corte, de coloração azulada com lobulação perceptível. As principais alterações histológicas foram verificadas no fígado, sob forma de necrose coagulativa e tumefação e/ou microvacuolização citoplasmática dos hepatócitos, e no rim representadas por acentuada nefrose, associada a grande quantidade de filtrado e/ou hemoglobina nos espaços de Bowman dentro de túbulos e do citoplasma das células epiteliais.

Copper(II) and Nickel (II) Complexation Capacity of Dissolved Organic Matter from Rivers of Agricultural and Urban Areas in the State of Sao Paulo

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Estudo morfofuncional dos rins de cães da raça Golden Retriever afetados pela distrofia muscular (GRMD)

A Distrofia Muscular de Duchenne (DMD) é uma miopatia severa de caráter recessivo ligada ao cromossomo X e o modelo animal de estudo mais relevante é o Golden Retriever Muscular Dystrophy (GRMD). Além das severas alterações que ocorrem na musculatura estriada, muitos estudos mostram que outras estruturas, inclusive viscerais, podem se mostrar alteradas nesta patologia. Desta forma, este trabalho objetivou análisar e comparar possíveis alterações estruturais e funcionais do rim em cães GRMD. Neste modelo de estudo, foi possível observar a presença das faces convexa e côncava, do hilo renal e dos pólos craniais e caudais dos rins. O órgão mostrou-se envolto por uma cápsula fibrosa. Em um corte sagital do órgão, notou-se a presença das regiões cortical e medular e da pelve renal. Na análise microscópica foi possível identificar a zona medular e cortical com suas estruturas: os corpúsculos renais formados pelo glomérulo e pela cápsula de Bowman, os túbulos contorcidos proximais e distais, os ductos coletores, vasos sanguíneos e os segmentos das Alças de Henle. As dosagens séricas de creatinina e uréia encontram-se dentro dos limites de normalidade. Desta forma, de acordo com os nossos resultados, podemos concluir que os animais afetados estudados, não apresentaram alterações estruturais ou funcionais dos rins, o que nos permitir sugerir que apesar da ingestão hídrica comprometida, a estrutura renal, mantem- se preservada nos animais GRMD.

La qualità erogata da un'azienda di trasporto collettivo in ambito urbano: calcolo del livello di servizio delle principali linee di ATC (BO)

Oggetto di questa tesi è lo studio della qualità del servizio di trasporto erogato che condiziona la qualità percepita dall’utente, poiché spesso proprio a causa di un errato processo di pianificazione e gestione della rete, molte aziende non sono in grado di consolidare un alto livello di efficienza che permetta loro di attrarre e servire la crescente domanda. Per questo motivo, si è deciso di indagare sugli aspetti che determinano la qualità erogata e sui fattori che la influenzano, anche attraverso la definizione di alcuni indicatori rappresentativi del servizio erogato. L’area di studio considerata è stata quella urbana di Bologna, e sono state prese in esame due linee di ATC, la 19 e la 27, caratterizzate entrambe da una domanda di trasporto molto elevata. L’interesse è ricaduto in modo particolare sugli aspetti legati alla regolarità del servizio, ovvero al rispetto della cadenza programmata delle corse e alla puntualità, ossia il rispetto dell’orario programmato delle stesse. Proprio da questi due aspetti, infatti, dipende in larga misura la percezione della qualità che gli utenti hanno del servizio di trasporto collettivo. Lo studio è stato condotto sulla base di dati raccolti attraverso due campagne di rilevamento, una effettuata nel mese di maggio dell’anno 2008 e l’altra nel mese di settembre dello stesso anno. La scelta del periodo, della zona e delle modalità di rilevamento è strettamente connessa all’obiettivo prefissato. Il servizio è influenzato dalle caratteristiche del sistema di trasporto: sia da quelle legate alla domanda che da quelle legate all’offerta. Nel caso della domanda di trasporto si considera l’influenza sul servizio del numero di passeggeri saliti e del tempo di sosta alle fermate. Nel caso dell’offerta di trasporto si osservano soprattutto gli aspetti legati alla rete di trasporto su cui si muovono gli autobus, analizzando quindi i tempi di movimento e le velocità dei mezzi, per vedere come le caratteristiche dell’infrastruttura possano condizionare il servizio. A tale proposito è opportuno dire che, mentre i dati della prima analisi ci sono utili per lo studio dell’influenza del tempo di sosta sull’intertempo, nella seconda analisi si vuole cercare di effettuare ulteriori osservazioni sull’influenza del tempo di movimento sulla cadenza, prendendo in esame altri elementi, come ad esempio tratti di linea differenti rispetto al caso precedente. Un’attenzione particolare, inoltre, verrà riservata alla verifica del rispetto della cadenza, dalla quale scaturisce la definizione del livello di servizio per ciò che riguarda la regolarità. Per quest’ultima verrà, inoltre, determinato anche il LOS relativo alla puntualità. Collegato al problema del rispetto della cadenza è il fenomeno dell’accodamento: questo si verifica quando i mezzi di una stessa linea arrivano contemporaneamente ad una fermata uno dietro l’altro. L’accodamento ha, infatti, origine dal mancato rispetto della cadenza programmata tra i mezzi ed è un’evidente manifestazione del mal funzionamento di un servizio di trasporto. Verrà infine condotta un’analisi dei fattori che possono influenzare le prestazioni del servizio di trasporto pubblico, così da collocare i dati ottenuti dalle operazioni di rilevamento in un quadro più preciso, capace di sottolineare alcuni elementi di criticità e possibili rapporti di causalità.

Response of Microcystis aeruginosa PCC 7806 to different CO2 concentrations

Future climatic change scenarios predict rising of the atmospheric CO2 levels which could favor the proliferation of some harmful bloom-forming cyanobacteria as Microcystis aeruginosa. In the present study, the response of M. aeruginosa strain PCC 7806 to two different partial pressure of CO2 was tested. Sandrini et al. (2013) recently found that several, but not all, M. aeruginosa strains lack the SbtA or BicA HCO3- uptake system genes; the contribution of different Ci transporters to photosynthesis and the difference between low and high affinity activated Ci uptake state were investigated. M. aeruginosa PCC 7806 was cultured in four chemostats containing modified BG11 medium with 10 mM NaNO3 and no presence of NaCl, NaHCO3, Na2CO3 and additional buffers. A wide variety of analysis on samples collected from continuous cultures – such as A750, medium composition, cellular composition, cell counting, mini-PAM, measurements with the O2 optode, Aminco, 77K fluorescence emission spectra – was carried out. Data analysis results showed that the increased CO2 concentration has a big effect on M. aeruginosa PCC 7806. Experiments were performed using the Oxy-4 O2 optode apparatus in order to measure the photosynthetic O2 evolution of samples taken from both batch and chemostat cultures. At low bicarbonate concentration, an evident inhibition of Na+-dependent HCO3- transporter BicA by LiCl at 25 mM was observed. The consequent addition of 25 mM NaCl was able to counteract the Li+ effect at pH 8.0 but not at pH 10.0. In the latter case, only the addition of a higher amount of HCO3- led to photosynthetic O2 evolution suggesting the important role of the BicA transporter. However, further studies are needed to better explain the results obtained as high pH levels might have an influence on the transport systems, altering the mechanism of pH regulation and the functioning of Na+/H+ antiporter systems.

The Presence of Words in Visual Art from Surrealism to Conceptualism

Without a doubt, one of the biggest changes that affected XXth century art is the introduction of words into paintings and, in more recent years, in installations. For centuries, if words were part of a visual composition, they functioned as reference; strictly speaking, they were used as a guideline for a better perception of the subject represented. With the developments of the XXth century, words became a very important part of the visual composition, and sometimes embodied the composition itself. About this topic, American art critic and collector Russell Bowman wrote an interesting article called Words and images: A persistent paradox, in which he examines the American and the European art of the XXth century in almost its entirety, dividing it up in six “categories of intention”. The aforementioned categories are not based on the art history timeline, but on the role that language played for specific artists or movements. Taking inspiration from Bowman's article, this paper is structured in three chapters, respectively: words in juxtaposition and free association, words as means of exploration of language structures, and words as means for political and personal messages. The purpose of this paper is therefore to reflect on the role of language in contemporary art and on the way it has changed from artist to artist.

Anticancer drug vinblastine sulphate induces transient morphological changes on the olfactory mucosa of the rabbit

Vinblastine sulphate (VBS) is an anticancer drug that acts by disrupting microtubule dynamics of highly mitotic tissue cells. The consequences of VBS on the olfactory mucosa (OM), a tissue with high mitotic numbers, are not clearly understood. We used qualitative and quantitative methods to determine the structural changes that may be produced on the rabbit OM by VBS. Following a single dose (0.31 mg/kg) of this drug, the structure of the mucosa was greatly altered on the first 3-5 days. The alteration was characterized by disarrangement of the normal layering of nuclei of the epithelia, degeneration of axonal bundles, occurrence of blood vessels within the bundles, localized death of cells of Bowman's glands and glandular degeneration. Surprisingly on or after day 7 and progressively to day 15 post-exposure, the OM was observed to regenerate and acquire normal morphology, and the vessels disappeared from the bundles. Relative to control values, bundle diameters, olfactory cell densities and cilia numbers decreased to as low as 53.1, 75.2 and 71.4%, respectively, on day 5. Volume density for the bundles, which was 28.6% in controls, decreased to a lowest value of 16.8% on day 5. In contrast, the volume density for the blood vessels was significantly lower in controls (19.9%) than in treated animals at day 2 (25.8%), day 3 (34.3%) and day 5 (31.5%). These findings suggest that the changes induced on the rabbit OM by VBS are transient and that regenerative recovery leads to the restoration of the normal structure of the mucosa.

A BAYESIAN HIERARCHICAL FRAMEWORK FOR SPATIAL MODELING OF fMRI DATA

Functional neuroimaging techniques enable investigations into the neural basis of human cognition, emotions, and behaviors. In practice, applications of functional magnetic resonance imaging (fMRI) have provided novel insights into the neuropathophysiology of major psychiatric,neurological, and substance abuse disorders, as well as into the neural responses to their treatments. Modern activation studies often compare localized task-induced changes in brain activity between experimental groups. One may also extend voxel-level analyses by simultaneously considering the ensemble of voxels constituting an anatomically defined region of interest (ROI) or by considering means or quantiles of the ROI. In this work we present a Bayesian extension of voxel-level analyses that offers several notable benefits. First, it combines whole-brain voxel-by-voxel modeling and ROI analyses within a unified framework. Secondly, an unstructured variance/covariance for regional mean parameters allows for the study of inter-regional functional connectivity, provided enough subjects are available to allow for accurate estimation. Finally, an exchangeable correlation structure within regions allows for the consideration of intra-regional functional connectivity. We perform estimation for our model using Markov Chain Monte Carlo (MCMC) techniques implemented via Gibbs sampling which, despite the high throughput nature of the data, can be executed quickly (less than 30 minutes). We apply our Bayesian hierarchical model to two novel fMRI data sets: one considering inhibitory control in cocaine-dependent men and the second considering verbal memory in subjects at high risk for Alzheimer’s disease. The unifying hierarchical model presented in this manuscript is shown to enhance the interpretation content of these data sets.