Dos peixes peçonhentos encontrados no Brasil os bagres destacam-se pelo número de acidentes provocados. O objetivo deste trabalho foi caracterizar os componentes tóxicos presentes na peçonha e no muco. Obtivemos uma média protéica de 3,1 mg/mL (peçonha) e 1,4 mg/mL (muco). O perfil eletroforético da peçonha e do muco possui poucas bandas protéicas. O fracionamento isolou 11 frações para peçonha e 13 frações para o muco. Com relação às atividades biológicas avaliadas, as frações peptídicas induziram danos teciduais ao passo que as frações protéicas induziram processo inflamatório. Isolamos duas frações com atividade antimicrobiana para cada secreção. Isolamos uma toxina de 65,1 kDa que apresenta homologia com Wap65 e análise por microscopia intravital revelou que esta proteína causa aumento dos leucócitos rolantes assim como a presença de leucócitos aderidos ao endotélio. Nossos resultados indicam uma diferença entre os componentes protéicos e peptídicos do muco e da peçonha. Finalmente, conseguimos isolar e caracterizar a proteína Wap65 da peçonha deste peixe. / Of the venomous fish found in Brazil, catfish noteworthy for the number of accidents they cause. The objective of this study was to characterize the toxic compounds present in the venom and mucus. We obtained an average protein intake of 3,1 mg / mL (venom) and 1,4 mg / mL (mucus). The electrophoretic profile of venom and mucus has a few protein bands. Fractionation isolated 11 fractions to the venom and 13 fractions to the mucus. With respect to biological activities evaluated, the peptide fractions induced tissue damage while the protein fractions induced inflammation. We isolated two fractions with antimicrobial activity for each sample. We isolated a toxin of 65,1 kDa which shows homology with Wap65 and intravital microscopy analysis revealed that this protein causes an increase in leukocyte rolling as well as the presence of leukocytes adhered to the endothelium. Our results indicate a difference between the peptide and protein components of mucus and venom. Finally, we isolate and characterize the protein Wap65 from the venom of this fish.

Submitted by Nadir Basilio (nadirsb@uninove.br) on 2016-05-17T20:42:43Z No. of bitstreams: 1 Gabriela Gomes Cardoso Rodrigues.pdf: 623885 bytes, checksum: 3da3d1230fd77beffc9c12f00be889bb (MD5) / Made available in DSpace on 2016-05-17T20:42:43Z (GMT). No. of bitstreams: 1 Gabriela Gomes Cardoso Rodrigues.pdf: 623885 bytes, checksum: 3da3d1230fd77beffc9c12f00be889bb (MD5) Previous issue date: 2015-02-04 / Chronic kidney disease (CKD) is characterized by progressive and irreversible loss of renal function and often progresses with a muscular weakness, whose set of signs and symptoms is generally referred to as uremic myopathy. Possible risk factors for the uremic myopathy are the uremic toxins. Among uremic toxins, indoxyl sulfate (IS) is a derivative of tryptophan metabolism by intestinal bacteria. Because skeletal muscle tissue undergo constant remodeling due differentiation of myoblasts in myotubes, it is possible that uremic toxins have a deleterious effect to influence this process, exacerbating the uremic myopathy. Low level laser therapy (LLLT) is regarded as a growth promoter feature widely used in the treatment of chronic diseases and has shown positive effects on the modulation of skeletal muscle repair process and also in the process of inflammation. However, in the context of CKD, the LLLT has not yet been explored. The aim of this study was to evaluate the effects of the IS on cell viability and on oxidative stress on cellular differentiation in cultured C2C12 myoblasts. In addition, to verify the action of the LLLT as a protective alternative to the cells. The C2C12 myoblasts were cultured in DMEM culture medium containing 10% fetal bovine serum and were induced to differentiation process by adding 2% horse serum. Three different IS concentrations were used to mimic the plasma concentrations of normal individual, CKD patients with moderate uremia and CKD patients with advanced uremia (0.6 mg/l and 53 mg/l and 236 mg/l, respectively), at different times of incubation (24 h, 48 h and 72 h). Subsequently, the cells were subjected to treatment with LLLT GaAlAs 780 nm (output power 10 mW, 20 seconds application time and energy density of 0.5 J / cm2). In terms of analysis, we used MTT method to assess the viability of the cells, flow cytometry to assess the viability/cell death and oxidative stress, nitrite dosing to evaluate nitric oxide production and real-time PCR to analyze IL-6, myogenin and MyoD expression (inflammation and cell differentiation markers). The results demonstrate that the IS at the maximum concentration was toxic to C2C12 cells, because it significantly decreased cell viability by MTT and by flow cytometry and by increasing the percentage of necrosis. This effect was present throughout the three incubation periods. With respect to oxidative stress, was not any conclusion, probably by the time the samples, but do not rule out the possibility of IS induce this type of stress. Although the IS has induced death to C2C12 cells, the remaining had no change in cell differentiation markers. Treatment with BPL the IS sensitized cells, reducing cell viability. We conclude that the IS acts directly on C2C12 myoblasts with toxic effect and may be the one factor responsible for uremic myopathy. Treatment with LLLT was not effective in protecting the cells. / A doença renal crônica (DRC) é caracterizada pela perda progressiva e irreversível da função renal e que frequentemente cursa com um quadro de fraqueza muscular, cujo conjunto de sinais e sintomas é globalmente designado como miopatia urêmica. Possíveis fatores predisponentes para a miopatia urêmica são as toxinas urêmicas. Dentre as toxinas urêmicas, o indoxil sulfato (IS) é uma derivada do metabolismo do triptofano presente em bactérias intestinais. Devido ao fato do tecido muscular esquelético sofrer constante remodelação graças à diferenciação de mioblastos em miotubos, é possível que toxinas urêmicas tenham um efeito deletério por influenciar este processo, agravando a miopatia urêmica. A terapia a laser de baixa potência (LBP) é considerada como um recurso bioestimulante amplamente utilizado no tratamento de doenças crônicas e tem demonstrado efeitos positivos sobre a modulação do processo de reparo muscular esquelético e também no processo da inflamação. Entretanto, no contexto de DRC, o LBP não foi ainda explorado. O objetivo do presente estudo foi avaliar dos efeitos do IS sobre a viabilidade celular, sobre o estresse oxidativo e sobre a diferenciação celular em cultura de mioblastos C2C12. Além disso, verificar a ação do LBP como forma de proteção às células. Os mioblastos C2C12 foram cultivados em meio de cultura de DMEM, contendo 10% de soro fetal bovino e foram induzidos ao processo de diferenciação por meio da adição de 2% soro de cavalo. Três diferentes concentrações de IS foram usadas para mimetizar as concentrações plasmáticas de indivíduo normal, paciente DRC com uremia moderada e paciente DRC com uremia avançada (0,6 mg/l; 53 mg/l e 236 mg/l, respectivamente), em diferentes períodos de incubação (24 h, 48 h e 72 h). Posteriormente, as células foram submetidas ao tratamento com laser de baixa potência AsGaAl 780 nm (potência de saída de 10 mW, tempo de aplicação de 20 segundos e densidade de energia de 0,5 J/cm2). Como análise, foi utilizado o método MTT para acessar a viabilidade das células, citometria de fluxo para avaliar a viabilidade/mortalidade das células, bem como o estresse oxidativo, dosagem de nitrito para avaliar a produção de óxido nítrico e PCR em tempo real para analisar a expressão de IL-6, miogenina e MyoD (marcadores de inflamação e diferenciação celular). Os resultados demonstram que o IS na concentração máxima foi tóxico para as células C2C12, pois diminuiu significativamente a viabilidade das células, tanto por MTT como por citometria de fluxo, aumentando a porcentagem de necrose. Este efeito foi presente nos três períodos de incubação. Com relação ao estresse oxidativo, não foi possível nenhuma conclusão, provavelmente pelo tempo das amostras , porém não descartamos a possibilidade do IS induzir este tipo de estresse. Embora o IS tenha induzido morte às células C2C12, as remanescentes não tiveram alteração dos marcadores de diferenciação celular. O tratamento com LBP sensibilizou as células ao IS, diminuindo a viabilidade das células. Concluímos que o IS age diretamente sobre mioblastos C2C12 com efeito tóxico, podendo ser um dos responsáveis pela miopatia urêmica. O tratamento com LBP não foi eficiente na indução de proteção às células.

246 samples of bulk and packaged spices from retail stores in the western, southeastern, southern, midwestern, and northeastern areas of the U.S. were examined for the presence of Clostridium -perfringens. Isolates were checked for the presence of the lecithinase gene (cpa) and enterotoxin genes (cpe) by PCR. Enterotoxin formation during sporulation was investigated using the Oxoid Toxin Detection Kit. Forty-three confirmed isolates (from 17% of total samples) were cpa-positive. Of those, 27 were cpe-positive. Together, levels of C. perfringens spores ranged from 3.6-2400/gm. The amount of enterotoxin in cell extracts ranged from 2-16 ng/ml. Some of the SEM images of isolated spore (# 78) and one plasmid-borne ent control (FD-153) showed an organized surface structure termed “candy-wrapper”. This extracellular structure remained after treatment with 0.1 % SDS for 1 hr, suggesting it was not composed of membrane debris from the mother cell. The D values of spores ranged from 1.19- 3.31 min. The addition of lysozyme in the plating medium elevated the recovery rate of heat-treated spores. The growth rate of a cocktail of spores from spices (# 31, # 32, # 45) between 4 to 5 hr after inoculation was determined with a doubling time of 6.82 min in hamburger. A cocktail of spores of plasmid-borne ent control showed an optimum growth rate between 5 to 8 hr after inoculation with doubling time of 15.98 min. However; spice isolate cocktail, plasmid-borne ent control cocktail (FD-5603 and FD-153), and a chromosome-borne ent control (NTCT 8239) were unable to germinate and outgrowth at 20oC. Inoculation in laboratory medium FTG indicated the same result as hamburger at 20oC. The ability of C. perfringens spores in spices to potentially survive cooking procedures can be followed by germination and growth of vegetative cells during improper cooling to levels associated with foodborne illness caused by this organism. Our results suggest that retail spices are potential vehicles of transmission of enterotoxin-positive C. perfringens.

Bacillus subtilis is a gram positive soil bacterium that is surrounded by many other microorganisms its environment. That is why it is necessary for the bacterium to be able to fight with these microorganisms for the nutrients and living space. B. subtilis contains the modules in its genetic make-up that improve its ability to compete. These modules are called the toxin-antitoxin systems. This Diploma Thesis is trying to identify yet undescribed extracellular toxins produced by the wild type BSB1 strain of B. subtilis. The related microorganism Bacillus megaterium was used as a competing bacterium. The contact-dependent or independent manner of killing the competing bacterium was demonstrated using this model. By deletion analysis and comparisons of the genomes of the various strains of B. subtilis, the SPβ prophage was first identified as a region containing an unknown toxin(s). Analysis of the extracellular proteome of B. subtilis subsequently revealed an unknown toxin (or toxin complex, respectively) of the molecular weight exceeding 100 kDa. Even more fascinating was the finding that such a large protein molecule is resistant to the pancreatic protease, trypsin. Subsequent non-enzymatic cyanogen bromide cleavage of the extracellular proteins and their analysis by mass spectrometry revealed...

Pore forming toxins (PFTs) are a large group of proteins found mainly in bacteria with some exceptions found in animals. They bind and form pores in their target membranes and form pores, which leads to cell death. Among these are cholesterol-dependent cytolysins (CDC), which require the presence of cholesterol to bind target membranes. Mitilysin (Mly), a protein of interest in this project, belongs to the CDC group of pore forming toxins. It is produced by the bacterium Streptococcus mitis, a pathogen closely related to Streptococcus pneumoniae, found in human oral cavity, which causes several diseases such as Viridans Group Streptococcal (VGS) toxic shock syndrome and endocarditis. Mly is a homologue of the toxin Pneumolysin, which is produced by S. pneumoniae. However, the mechanism of pore formation is not well known. The purpose of this project is to understand the mechanism of CDC pore formation, focusing on the key amino acid residues that are responsible for transitioning from Mly pre-pore to pore state. The findings will aid in the design of inhibitors of pore formation as potential anti-bacterial drug candidates. The major goal of the project was to determine the 3-dimensional (3D) structure of assembled Mly pore. Mly is expressed in E.coli and purified by Ni-NTA affinity chromatography. Pore formation is confirmed by a hemolysis assay and negative stain-transmission electron microscopy. Mly pores are vitrified, analyzed and imaged in a cryo-electron microscope. 2D images were processed to generate a 3D density map. However, our Mly pore 3D map was incomplete due to lack of 2D projection angles resulting from preferred orientation of pore particles during sample preparation. To overcome this problem, we aim to use DNA origami, which requires His-tagged Mly. We were able to determine that His-tagged Mly retains its pore formation ability.

Patients with kidney failure (KF) have a high incidence of cardiovascular (CV) disease, partly driven by insufficient clearance of uremic toxins. Recent investigations have questioned the accepted effects of adverse lipid profile and CV risk in uremic patients. Therefore, we related a panel of uremic toxins previously associated with CV morbidity/mortality to a full lipid profile in a large, tri-national, cross-sectional cohort. Total, high-density lipoprotein (HDL), non-HDL, lowdensity lipoprotein (LDL), and remnant cholesterol, as well as triglyceride, levels were associated with five uremic toxins in a cohort of 611 adult KF patients with adjustment for clinically relevant covariates and other patient-level variables. Univariate analyses revealed negative correlations of total, non-HDL, and LDL cholesterol with all investigated uremic toxins. Multivariate linear regression analyses confirmed independent, negative associations of phenylacetylglutamine with total, non-HDL, and LDL cholesterol, while indole-3 acetic acid associated with non-HDL and LDL cholesterol. Furthermore, trimethylamine-N-Oxide was independently and negatively associated with non-HDL cholesterol. Sensitivity analyses largely confirmed findings in the entire cohort. In conclusion, significant inverse associations between lipid profile and distinct uremic toxins in KF highlight the complexity of the uremic milieu, suggesting that not all uremic toxin interactions with conventional CV risk markers may be pathogenic.

No description available.

Nordic Nature is unique with long coastline, deep forest, mountains, rivers, and valleys, fromthe Arctic tundra of North to warm parts in south. Climate change is one of the major problemsthe Nordic countries are facing. The threat climate change is creating on the environment hascreated a consequence in Health, lifestyles, diverse society, and a whole ecosystem. Theseenvironmental toxins are creating significant risk on human health as well as wellbeing causingvarious acute, Chronic as well as autoimmune diseases. In this recent time Nordic Countrieshave been experiencing forest fires, floods, droughts, precipitation, hotter, colder, and windydays. The climate in recent years has been showing some noticeable variations. The Nordiccountries are considered as the leader in climate change moderations, but the above issues arecreating changes and slowly hampering health and lifestyles of people living there. Thesehuman induced climate changes are already creating many health problems such as allergies,asthma, many other respiratory problems, giardiasis, encephalitis, cardiovascular diseases,endocrine diseases, fertility issues and autoimmune diseases. WHO (World HealthOrganization) is also concern about the rise in the temperature in Artic region as result ofclimate change and sharing measures to prevent from these hot flushes. This thesis topic usuallyshows the connection between climate change and environmental toxins. Moreover, itunderlines how these toxins create impacts on human health and measures taken by Nordiccountries to overcome it. Thus, in this thesis we are using integrative literature review to studyhow climate change and environmental toxins are creating human health and lifestyle in Nordicregion. We took a few cases of scientific journals and articles which show how climate changeand environmental creating hazardous diseases, creating environmental pollution, and howNordic countries are taking initiatives to control them. During the research ethicalconsideration was also followed strictly. We wanted to highlight this topic to make peopleaware as climate change and environmental toxins do not look like serious issues for now, butit is like “Slow Poisons.”

Sammanfattning Dammutrivningar blir alltmer vanligt på grund av att många är föråldrade och kostar mer att driva och underhålla än vinsten de ger i energi. Effekten av utrivningar på halten miljögifter i fiskar är relativt okänd, men det finns observationer som indikerar att det dammutrivningar kan minska kvicksilverhalten i fiskar. Syftet med min studie var att undersöka skillnaderna i kvicksilverhalten hos gädda (Esox lucius), mellan sjöar som har fått dammen i sjöutloppet utriven respektive dammen intakt. Jag undersökte 66 provtagningar från 18 oligotrofa svenska skogssjöar. I sjöar där dammutrivning skett innehöll en medelstor gädda 500 ng/g kvicksilver, medan motsvarande värde i sjöar med dammen intakt var 740 ng/g kvicksilver. Det fanns ingen signifikant effekt av vare sig antal år efter utrivning eller provtagningsår. Kvicksilverhalten var nära förknippad med gäddans längd och varierade mellan sjöar, och i fyra av sjöarna, översteg mätningarna 1000n ng/g (gränsvärdet för konsumtion inom EU) medan gäddor från de andra sjöarna hade värden som mestadels under detta gränsvärde. / Abstract Dam removals are becoming increasingly common because many are outdated and cost more to operate and maintain than the value of that they provide in energy. The effect of dam removal on the level of environmental toxins in fish is relatively unknown, but there are observations that indicate that dam removal can reduce mercury concentrations in fish. The aim of my study was to investigate differences in the mercury concentration of pike (Esox lucius) between lakes that have had their outlet dams removed and those that have their dams intact. I examined 66 samples from 18 oligotrophic Swedish forest lakes.n lakes where the dam had been removed, a pike of average length contained 500 ng/g mercury, whereas the corresponding value for lakes with the dam intact was 740 ng/g of mercury. There was no significant effect of the number of years after removal or sampling year. The mercury concentration was closely associated to pike length and varied between the lakes, and in four of the lakes, all measurements exceeded 1000 ng/g (the limit value for consumption in EU), whereas most pike from the other lakes had values below this limit value.

Thesis (M. Tech. Environmental health) -- Central University of technology, Free State, 2011 / In nature microorganisms do not exist alone, but in association with one another. These kinds of associations can also be found in food industries, where cells of the same or different species can attach to pipes (biofilm formation) and a variety of surfaces in food processing environments and in food product such as yoghurt which can contain both yeast and bacteria originating from the starter culture as well as fruit. To control food spoilage organisms and food-borne pathogens preventative measures such as good manufacturing processes, the use of sanitizers and preservatives as well as hazard analysis critical control points (HACCP) are crucial in food industries. Sanitation of the working surface, floors, pipes, containers and equipment is a stepwise application of a detergent, acid or alkali rinse, a disinfectant treatment followed by final rinsing. If rinsing of the sanitizer is not done properly it may end up in the product in sub-lethal doses. In this study the influence of Liquid Hypochlorite (LH) and Liquid Iodophore (LI) sanitizers on organism growth and toxicity was evaluated. The organisms investigated included Escherichia coli 0113, Escherichia coli 026 and Zygosaccharomyces bailii Y-1535 in yeast malt broth, which was supplemented with LH and LI at sub-lethal concentrations 0.05% LH, 0.2% LH and 0.075% LI. Subsequently, bacterial and yeast growth responses as pure cultures and in combination (E. coli + Z. bailii) were measured as colony forming units and optical density values. Incorporation of the sanitizers in the growth media resulted in different levels of growth inhibition. Z. bailii proved more robust and the growth rate was not influence significantly by the addition of sanitizers or communal growth with either E. coli strains. The growth rate of both E. coli strains decreased where grown in combination with Z. bailii as well as in the presence of sanitizers, with the most influence exerted by LH. Changes in endotoxicity following the growth of the test samples (stressed cells) and the control (unstressed) were measured by the limulus amoebocyte lysate (LAL) and porcine IL-6 ELISA methods. Where E. coli strains were cultured together with Z. bailii the toxicity of tire mixture showed a decrease over time when measured with the limulus amoebocyte assay method. Interestingly the communal growth of the E. coli strains and Z bailii produced different toxicity profiles when the IL-6 porcine method was used, hi both cases, where E. coli strains were cultured together with Z. bailii the toxicity of the mixture showed an increase over tune when measured by this assay. Other than a similar toxicity profile for E. coli 0113 grown in pure culture, the comparison between results obtained using the LAL or porcine IL-6 methods yielded no correlation in determined toxicity. It was established that LH and LI sanitizers as well as communal growth had an influence in the toxicity of LPS/EPS and the method used to determine such toxicity should be carefully considered.