Estudo electroquímico de compostos naturais e semicondutores com possível utilização em células foto voltaicas

As células foto voltaicas orgânicas ou células de Gräetzel (depois do seu descobridor) são aparelhos para a colecta de energia solar que utilizam um semicondutor inorgânico e uma molécula orgânica. Dita molécula orgânica é capaz de excitar-se na presença de radiação electromagnética e ceder esta energia através da doação de electrões a este semicondutor. Embora estas estruturas e o seu processo de fabrico sejam relativamente pouco onerosas, o aproveitamento da energia solar é ainda muito baixo. Para além desta deficiência, os corantes sintéticos sofrem de “bleaching” ou então são reduzidos ou oxidados facilmente quando não conseguem transferir a energia que foi absorvida ou quando é difícil voltar ao estado original por dificuldades no completamento de circulação de electrões. Neste trabalho pretende-se então estudar o comportamento de moléculas e misturas complexas de moléculas com capacidade para serem excitadas pela luz solar. Como a dita xcitação promove a transferência de um electrão, este processo será seguido pela técnica de Voltametria cíclica. Como substâncias absorventes de luz utilizaremos compostos naturais (principalmente flavonóides) puros, ou então na forma de complexos naturais extraídos de algumas plantas. Estas misturas de corantes serão extractos aquosos (infusões) de casca de laranja e limão assim como extractos de folhas de cerejeira, com o objectivo de proporcionar lternativas aos flavonóides utilizados neste estudo. A caracterização voltamétrica desta célula é feita em diferentes formas de iluminação. Sobre a célula assim formada faz-se incidir rimeiro luz de lâmpadas fluorescentes, depois luz ultra violeta e por fim sem qualquer tipo de luz incidente. Na base do fabrico da variante mais clássica destas células está o semicondutor óxido de itânio (TiO2), por ser uma substância muito comum e barata e com propriedades semicondutoras notáveis. Uma forma comum de melhorar a eficiência deste material é introduzir dopantes com o intuito de melhorar a eficiência do processo de transferência electrónica. Um segundo objectivo deste trabalho é o estudo de sistemas semicondutor/molécula foto activa. Semicondutores como ZnO, TiO2 e TiO2 dopado serão então estudados. O gels de TiO2 ou o TiO2 dopado serão depositados sobre lâminas de vidro comum, nas quais foi anteriormente depositado uma película de alumínio que serve de condutor (eléctrodo egativo). Uma outra variante será a utilização de óxido de zinco, um semicondutor de baixo custo que por sua vez vai ser depositado em lâminas de alumínio comercial. A nossa célula foto electroquímica será então formada por moléculas de corante, uma lâmina e um semicondutor (que funcionará como eléctrodo de trabalho), com ou sem electrólito/catalizador (solução de iodo/iodeto), e eléctrodos de referência de Ag/AgCl, e outro auxiliar de grafite. Um outro objectivo é fazer um pequeno estudo sobre influencia do catalisador I2/etilenodiamina no comportamento electroquímico da célula, de forma a poder utilizar o solvente (etilenodiamina) com menor volatilidade do que a água, que é empregada no par I2/I3.m A importância deste facto prende-se com a limitada vida destas células quando o electrólito/solvente é evaporado pelas altas temperaturas da radiação incidente.

Leishmania infantum AS A CAUSATIVE AGENT OF CUTANEOUS LEISHMANIASIS IN THE STATE OF MATO GROSSO DO SUL, BRAZIL

Cutaneous leishmaniasis is caused by different species of theLeishmania genus. Leishmania(Leishmania) infantum, causing cutaneous leishmaniasis, has been described in patients living in areas where visceral leishmaniasis is endemic. In this study, it was possible to characterize this species in seven slides from cutaneous tissue imprints from patients with cutaneous leishmaniasis in the State of Mato Grosso do Sul, Brazil.

Histopathology of human American cutaneous leishmaniasis before and after treatment

Chemical therapy for the treatment of leishmaniasis is still inadequate, and a number of drugs and therapeutic programs are being tested. Besides treatment, the ultimate goal is an effective cure, and histopathological analyses of the lesion cicatrices constitute an important measure of treatment success, or otherwise, in this respect. In this paper, we describe histopathological patterns in cases of American cutaneous leishmaniasis in 32 patients from the municipality of Caratinga, Minas Gerais, Brazil, before and after treatment with the following therapeutic methodos: l) leishvacin + glucantime; 2) leishvacin + BCG associated with glucantime; 3) glucantime; 4) leishvacin + BCG. Lesion fragments were collected from all patients by biopsy prior to, and approximately 30 days after, each treatment which resulted in a clinical diagnosis of cure. Following the analysis of slides, the preparations were described from a histopathological point of view and grouped taking into account the prevalence or significance of the characteristic elements. This process resulted in the following classification: 1. exsudative reaction (ER); 2. exsudative giant cell reaction (EGCR); 3. exsudative productive reaction (EPR); 4. exsudative productive giant cell reaction (EPGCR); 5. exsudative productive necrotic reaction (EPNR); 6. necrotic exsudative reaction (NER); 7. productive exsudative reaction (PER), 8. productive giant cell reaction (PGCR); 9. productive exsudative giant cell reaction (PEGCR); 10. productive exsudative giant cell granulomatous reaction (PEGCGR); 11. productive reaction (PR) and 12. productive cicatricial (cure) reaction (PCR). After this analysis, it was noted that clinical cure did not always coincide with histopathological cure.

Production of anti-Cryptosporidium polyclonal antibodies and standardization of direct immunofluorescence for detecting oocysts in water

INTRODUCTION: The production of anti-Cryptosporidium polyclonal antibodies and its use in direct immunofluorescence assays to determine the presence of Cryptosporidium in water are described in the present work. METHODS: Two rabbits were immunized with soluble and particulate antigens from purified Cryptosporidium oocysts. The sera produced were prepared for immunoglobulin G extraction, which were then purified and conjugated with fluorescein isothiocyanate (FITC). Slides containing known amounts of oocysts were prepared to determine the sensitivity of the technique. To test the specificity, slides containing Giardia duodenalis cysts were prepared. RESULTS: The conjugate was successfully used in water samples experimentally contaminated with Cryptosporidium oocysts, and it was possible to detect up to five oocysts/spot, corresponding to contamination of 250 oocysts/mL. CONCLUSIONS: The three immunizations performed in the rabbits were enough to produce antibodies against Cryptosporidium, the standard direct immunofluorescence assay permitted the detection of five oocysts in 20% of the samples, and no cross-reaction with Giardia duodenalis cysts occurred.

Increased detection of schistosomiasis with Kato-Katz and SWAP-IgG-ELISA in a Northeastern Brazil low-intensity transmission area

INTRODUCTION: The laboratory diagnosis of schistosomiasis is based mainly on the detection of parasite eggs in stool samples through the Kato-Katz (KK) technique, reading one slide by test. However, a widely known limitation of parasitological methods is reduced sensitivity, particularly in low endemic areas. METHODS: To increase sensitivity, we conducted further slide readings from the same stool sample using the parasitological method associated with a serological test. We used the KK method (three slides) and the IgG anti-Schistosoma mansoni-enzyme-linked immunosorbent assay (ELISA) technique to diagnose schistosomiasis in low endemic areas in the Brazilian State of Ceará. Fecal samples and sera from 250 individuals were analyzed. RESULTS: Sixteen percent and 47.2% of samples were positive in parasitological tests and serological tests, respectively. Parasitological methods showed that 32 (80%) individuals tested positive on the first slide, 6 (15%) on the second slide, and 2 (5%) on the third. The performance of the ELISA test in the diagnosis, using the KK method as diagnostic reference, showed a negative predictive value of 100%, with specificity and positive predictive values of 62.8% and 33.9%, respectively. CONCLUSIONS: In this study, the increase from one to three slides analyzed per sample using the KK technique was shown to be a useful procedure for increasing the diagnostic sensitivity of this technique.

Occurrence of Cryptosporidium spp., Giardia spp. and other pathogenic intestinal parasites in the Beberibe River in the State of Pernambuco, Brazil

INTRODUCTION: Transmission of pathogenic protozoa and helminths by water is a serious public health problem. In this study, we analyzed the presence of these organisms in the Beberibe River in Pernambuco, Brazil. METHODS: Parasite analysis was performed using the Hoffman, Pons, & Janer method followed by centrifugation and preparation of slides by staining with acetic acid and Lugol's solution. Protozoan oocysts were isolated by the modified Ziehl Neelsen method. RESULTS: Cryptosporidium spp., Giardia spp. and other parasites were found in the Beberibe River. CONCLUSIONS: Sanitation companies must assess pathogenic intestinal parasites in water basins providing public water and subsequently develop improved treatment systems for removal of such parasites.

Polymerase chain reaction-based method for the identification of Leishmania (Viannia) braziliensis and Leishmania (Viannia) guyanensis in mucosal tissues conserved in paraffin

ABSTRACTINTRODUCTION: In the Americas, mucosal leishmaniasis is primarily associated with infection by Leishmania (Viannia) braziliensis. However, Leishmania (Viannia) guyanensis is another important cause of this disease in the Brazilian Amazon. In this study, we aimed at detecting Leishmaniadeoxyribonucleic acid (DNA) within paraffin-embedded fragments of mucosal tissues, and characterizing the infecting parasite species.METHODS: We evaluated samples collected from 114 patients treated at a reference center in the Brazilian Amazon by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analyses.RESULTS: Direct examination of biopsy imprints detected parasites in 10 of the 114 samples, while evaluation of hematoxylin and eosin-stained slides detected amastigotes in an additional 17 samples. Meanwhile, 31/114 samples (27.2%) were positive for Leishmania spp. kinetoplast deoxyribonucleic acid (kDNA) by PCR analysis. Of these, 17 (54.8%) yielded amplification of the mini-exon PCR target, thereby allowing for PCR-RFLP-based identification. Six of the samples were identified as L. (V.) braziliensis, while the remaining 11 were identified as L. (V.) guyanensis.CONCLUSIONS: The results of this study demonstrate the feasibility of applying molecular techniques for the diagnosis of human parasites within paraffin-embedded tissues. Moreover, our findings confirm that L. (V.) guyanensisis a relevant causative agent of mucosal leishmaniasis in the Brazilian Amazon.

Life cycle of Trypanosoma cruzi (y strain) in mice

Since 1958, we have studied experimental Chagas' disease (CD) by subcutaneous inoculation of 1,000 blood forms of Trypanosoma cruzi (Y strain) in Balb/C. mice. Evolution of parasitemia remained constant, beginning on the 5th and 6th day of the disease, increasing progressively, achieving a maximum on about the 30th day. After another month, only a few forms were present, and they disappeared from the circulation after the third month, as determined from direct examination of slides and the use of a Neubauer Counting Chamber. These events coincided with the appearance of amastigote nests in the tissues (especially the cardiac ones), starting the first week, and following the Gauss parasitemia curve, but they were not in parallel until the chronic stage. In 1997, we began to note the following changes: Parasites appeared in the circulation during the first week and disappeared starting on the 7th day, and there was a coincident absence of the amastigote nests in the tissues. A careful study verified that young forms in the evolutionary cycle of T. cruzi (epi + amastigotes) began to appear alongside the trypomastigotes in the circulation on the 5th and 7th post-inoculation day. At the same time, rounded, oval, and spindle shapes were seen circulating through the capillaries and sinusoids of the tissues, principally of the hematopoietic organs. Stasis occurs because the diameter of the circulating parasites is greater than the vessels, and this makes them more visible. Examination of the sternal bone marrow revealed young cells with elongated forms and others truncated in the shape of a "C" occupying the internal surface of the blood cells that had empty central portions (erythrocytes?). We hypothesize that there could be a loss of virulence or mutation of the Y strain of Trypanosoma cruzi.

Histological comparison of the alar nasal cartilages in unilateral cleft lip

Patients with unilateral cleft lip display characteristic nasal changes that are independent of the degree of deformity. Defenders of the intrinsic theory consider these deformities to be due to embryogenic alterations of the alar nasal cartilages. Those that propose the extrinsic theory defend the thesis that the deformity is due to disorganization of the perioral muscles deformed by the cleft. The purpose of this study is to contribute histological evidence to help clarify the issue. PATIENTS AND METHODS: Specimens of the lateral portion of both the healthy and the cleft side of the alar cartilages were obtained from 18 patients. These uniformly cut specimens were stained by hematoxylin and eosin. Samples from 2 patients were excluded due to imperfections. The same pathologist examined all the slides. He was unaware of the origins of the specimens; he counted the number of chondrocytes and quantified the cartilage matrixes. RESULTS: All data was analyzed statistically, and no significant statistical differences were apparent, either in the number of chondrocytes or the cartilage matrix between the healthy side and the cleft side. DISCUSSION: These results apparently support the group that defend the extrinsic theory; nevertheless, the doubt about the composition of the cartilage matrix remains, not only concerning the glycosaminoglycans that compose them, but also regarding elastin and collagen and its linkages that can cause different degrees of collagen consistency.

Structural and successional variability of periphytic algal community in a Amazonian lake during the dry and rainy season (Rio Branco, Acre)

The colonization process and successional patterns of a periphytic algal community were evaluated in a Amazonian Viveiro Lake (Rio Branco, Acre, Brazil). Sampling was performed over a period of 35 days; at four-day intervals for 20 days, and then at five-day intervals. Water sampling for physical, chemical and biological analyses was done during the dry and rainy season. Glass slides were used as artificial substrates for periphyton colonization. The structural community was evaluated through population density, algae class, diversity indices and descriptive species. Species richness, diversity and evenness increased as succession progressed. While density of Bacillariophyceae, Euglenophyceae and Zygnemaphyceae increased with succession, Cyanobacteria remained dominant. Synechocystis aquatilis, Synechocystis diplococcus and Navicula pseudolanceolata were the main descriptive species in both the dry and rainy season. Cymbela tumida, Frustulia rhomboides, Trachelomonas lacustris and Closterium acicularis was correlated with an increase in hydrologic level during the rainy season. Conversely, the density of Chlamydomonas sp., Chroomonas nordstedtii, Trachelomonas volvocinopsis, Trachelomonas volvocina and Synechococcus linearis was correlated with an increase in water transparency during the dry season. In general, the periphytic algal community showed high diversity and species richness independent of season. Season also had little influence on representation of algae class and main descriptive species. However, successional patterns varied by season, and changes in hydrologic levels acted directly on the succession path of periphytic algae. More research on periphyton dynamics is needed to improve our understanding of tropical lake ecosystems, especially in Amazonian.

Comparing different methods to measure biofilm thickness: the techniques are: optical coherence tomography, confocal laser scanning microscopy and low load compression test

Dissertação de mestrado integrado em Engenharia Biomédica (área de especialização em Engenharia Clínica)

Fractal Dimension in Quantifying Experimental-Pulmonary-Hypertension-Induced Cardiac Dysfunction in Rats

Abstract Background: Right-sided heart failure has high morbidity and mortality, and may be caused by pulmonary arterial hypertension. Fractal dimension is a differentiated and innovative method used in histological evaluations that allows the characterization of irregular and complex structures and the quantification of structural tissue changes. Objective: To assess the use of fractal dimension in cardiomyocytes of rats with monocrotaline-induced pulmonary arterial hypertension, in addition to providing histological and functional analysis. Methods: Male Wistar rats were divided into 2 groups: control (C; n = 8) and monocrotaline-induced pulmonary arterial hypertension (M; n = 8). Five weeks after pulmonary arterial hypertension induction with monocrotaline, echocardiography was performed and the animals were euthanized. The heart was dissected, the ventricles weighed to assess anatomical parameters, and histological slides were prepared and stained with hematoxylin/eosin for fractal dimension analysis, performed using box-counting method. Data normality was tested (Shapiro-Wilk test), and the groups were compared with non-paired Student t test or Mann Whitney test (p < 0.05). Results: Higher fractal dimension values were observed in group M as compared to group C (1.39 ± 0.05 vs. 1.37 ± 0.04; p < 0.05). Echocardiography showed lower pulmonary artery flow velocity, pulmonary acceleration time and ejection time values in group M, suggesting function worsening in those animals. Conclusion: The changes observed confirm pulmonary-arterial-hypertension-induced cardiac dysfunction, and point to fractal dimension as an effective method to evaluate cardiac morphological changes induced by ventricular dysfunction.

Microdeterminação do arsênico

Studying the application of those methods of microanalysis which avoid costly instalations and atempting to combine high precision with low cost, the author recomends a new method consisting of the following : a) exposure of a surface of 530.66 mm2 of Zn to the action of the acid. b) instalation of 3 series of HgBr2 paper in test tubes with an internal diameter of respectively 3,5 and 9 mm. c - mouting between two slides, covering the margins (with parafin etc.) with parafin in order to conserve the results of the determination without change due to the action of light or moisture. d) the results can be compared at a level of 0.00001 mgr. A203 or 0.000007575 mgr. As.

Sobre o preparo dos parasitas da malária em distensões espessas, especialmente em amostras supersecas

I) the A. presents a method developed for the preparation of thick blood films, specially old desiccated smears. The observations are based on the experience of more than 53000 blood samples collected in the laboratory of the “Serviço de Malaria do Nordeste” as well as in the research department of the “Serviço de Malaria da Baixada Fluminenese”. II) As in introductory matter, he emphasizes the value of the obstacles presented by overdrying of the thick blood films occurring systematically in great malaria control organizations in which the laboratory receives materials from more or less remote localities, particularly in the Brazilian northeast, in regions invaded by Anopheles gambiae. III) An analysis of the causes of failure of the methods of Chorine and Knowles recorded in the literature for such purposes is given, as well as its adaptability for the simultaneous preparation of large numbers of samples. IV) The method is based the protective action of a previous fixation by a dilute solution of formalin, which, without preventing further dehemoglobinization, prevents morphological alterations in the parasites by the action of Knowles solution which is retained in this metod without modification. V) For washing out the acids of the dehemoglobinizating solution as well as for diluting the Giemsa stain, the A. proposes a very simple technique, extremely convenient for such purpose, which consists in adding acetic acid to the distilled water in the proportion of 1 drop for each 10cc of water, and then increasing the hydrogeni-on concentration to pH 7.2 with a 2% sol. of sodium carbonate. As indicator a 0.02% solution purple-bromcresol prepared in accordante to Medalia, is used. In this reaction there is the formation of the acetic acid ↔ sodium acetate, buffer system very suitable for giving a convenient pH and for preventing the precipitation of the dye, which can be used for two or three batches of 700 or 800 slides each, without changing the staining solution. VI) – The method can be summarized as follow: For a small number of samples, Coplin’s or any other staining jar can be used. Large number of slides must be placed in groups of 10 or 15 units each, the slides being separated by a piece of cardboard, according to Barber & Komp. A) Fix in dilute formalin (2%), during 5 minutes. b) Without washing, put in Knowles solution (see the formula in the text), no more than 20 minutes. c)Two successive washings in distilled water, buffered as explained above (which can be used several times). d) Dry and stain with Giemsa solution, prepared by using 1 drop of the stain for each c. c. of buffered distilled water. Time: 1 hour. E) Was in distilled water and dry.

Aspectos parasitários observados no local inoculado com esporozoitos de Plasmodium Gallinaceum

The following is a summary of the studies made on the development of Plasmodium gallinaceum sporozoites inoculated into normal chicks. Initially large numbers of laboratory reared Aëdes aegypti were fed on pullets heavily infected with gametocytes. Following the infectious meal the mosquitoes were kept on a diet of sugar and water syrup until the appearance of the sporozoites in the salivary glands. Normal chicks kept in hematophagous arthropod proof cages were then inoculated either by bite of the infected mosquitoes or by subcutaneous inoculations of salivary gland suspensions. By the first method ten mosquitoes fed to engorgement on each normal chick and were then sacrificed immediately afterwards to determine the sporozoite count. By the second method five pairs of salivary glands were dissected out at room temperature, triturated in physiological saline and inoculated subcutaneously. The epidermis and dermis at the site of inoculation were excised from six hours after inoculation to forty eight hours after appearance of the parasites in the blood stream and stretched out on filter paper with the epithelial surface downward. The dermis was then curretted. Slides were made of the scrapings consisting of connective tissue and epithelial cells of the basal layers which were fixed by metyl alcohol and stained with Giemsa for examination under the oil immersion lens. Skin fragments removed from normal chicks and from regions other than the site of inoculation in the infected chicks were used as controls. In these, only the normal histological aspect was ever encountered. In the biopsy made at the earliest period following inoculation clearly defined elongated forms with eight or more chromatin granules arranged in rosary formation were found. The author believes these to be products of the sporozoite evolution. Search for transition stages between these forms and sporozoites is planned in biopsies to be taken immediately following inoculation and at given intervals up to the six hour period. 1.) 6 and 12 hour periods. The bodies referred to above found in the first period in great abundance, apparently in proportion to the large numbers of sporozoites inoculated, were perceptibly reduced in numbers in the second period. 2.) 18 hour period. Only one biopsy was examined. This presented a binuclear body shown in Fig. 1, having a more or less hyaline protoplasm staining an intense blue and a narrow vacuole delimiting the cell boundaries. The two chromatin grains were quite large presenting a clearly defined nuclear texture. 3.) 24 hour period. A similar body to that above (Fig. 2) was seen in the only preparation examined. 4.) 60 hour period. The exoerythrocytic schizonts were found more frequently from this period onward. Several such were found no longer to contain the previously described vacuoles (Fig. 3). 5.) 84 hour period. Cells bearing eight or more schizonts were frequently encountered here. That these are apparently not bodies in process of division may be seen in Fig. 4. From this time onward small violet granules similar to volutine grains appeared constantly in the schizont nucleus and protoplasm. These are definitely not hemozoin. The above observations fell within the incubation period as repeated examinations of the peripheral and visceral blood were negative. Exoery-throcytic parasites also were never encountered in the viscera at this time. Exoerythrocytic schizonts searched for at site of inoculation 1, 24 and 48 hours after the incubation period were present in large number at all three times with apparent tendency to diminish as the number within the blood stream increased. Many of them presented the violet granules mentioned above. The appearance of the chromatin and the intensity of staining of the protoplasm varied from body to body which doubtless corresponds to the evolutionary stage of each. This diversity of aspect may frequently be seen in the parasites of the same host cell (Fig. 5.). These findings lend substance to the theory that the exoerythrocytic forms are the link between the sporozoites and the pigmented parasites of the red blood corpuscles. The explanation of their continued presence in the organism after infection of the blood stream takes place and their presence in cases infected by the inoculation blood does not come within the scope of this work. Large scale observations shortly to be undertaken will be reported in more detail particularly observations on the first evolutionary phases of the sporozoite within the organism of the vertebrate host.