Toxicity of linear alkylbenene sulphonate (LAS) detergent, to Clarias gariepinus fingerlings

The acute toxicity of Linear Alkylbenzene Sulphonate (LAS) detergent to Clarias gariepinus fingerlings was investigated using static bioassays and continous aeration over a period of 96h. The 96h LC sub(50) was determined as 24.00mgL super(-1). During the exposure period, the test fish exhibited several behavioural changes before death such as restlessness, rapid swimming, loss of balance, respiratory distress and haemorrhaging of gill filaments amongst others. Opercula ventilation rate as well as visual examination of dead fish indicates lethal effects of the detergent on the fish. Water quality examination showed increase in pH from 6.55 to the alkaline, death point of 10.55. There was also a remarkabel rise of alkalinity from 20.00mgL super(-1) to 52.50mgL super(-1)

Model systems for the study of drug hypersensitivity

I. It was not possible to produce anti-tetracycline antibody in laboratory animals by any of the methods tried. Tetracycline protein conjugates were prepared and characterized. It was shown that previous reports of the detection of anti-tetracycline antibody by in vitro-methods were in error. Tetracycline precipitates non-specifically with serum proteins. The anaphylactic reaction reported was the result of misinterpretation, since the observations were inconsistent with the known mechanism of anaphylaxis and the supposed antibody would not sensitize guinea pig skin. The hemagglutination reaction was not reproducible and was extremely sensitive to minute amounts of microbial contamination. Both free tetracyclines and the conjugates were found to be poor antigens.

II. Anti-aspiryl antibodies were produced in rabbits using 3 protein carriers. The method of inhibition of precipitation was used to determine the specificity of the antibody produced. ε-Aminocaproate was found to be the most effective inhibitor of the haptens tested, indicating that the combining hapten of the protein is ε-aspiryl-lysyl. Free aspirin and salicylates were poor inhibitors and did not combine with the antibody to a significant extent. The ortho group was found to participate in the binding to antibody. The average binding constants were measured.

Normal rabbit serum was acetylated by aspirin under in vitro conditions, which are similar to physiological conditions. The extent of acetylation was determined by immunochemical tests. The acetylated serum proteins were shown to be potent antigens in rabbits. It was also shown that aspiryl proteins were partially acetylated. The relation of these results to human aspirin intolerance is discussed.

III. Aspirin did not induce contact sensitivity in guinea pigs when they were immunized by techniques that induce sensitivity with other reactive compounds. The acetylation mechanism is not relevant to this type of hypersensitivity, since sensitivity is not produced by potent acetylating agents like acetyl chloride and acetic anhydride. Aspiryl chloride, a totally artificial system, is a good sensitizer. Its specificity was examined.

IV. Protein conjugates were prepared with p-aminosalicylic acid and various carriers using azo, carbodiimide and mixed anhydride coupling. These antigens were injected into rabbits and guinea pigs and no anti-hapten IgG or IgM response was obtained. Delayed hypersensitivity was produced in guinea pigs by immunization with the conjugates, and its specificity was determined. Guinea pigs were not sensitized by either injections or topical application of p-amino-salicylic acid or p-aminosalicylate.

On the toxicity of some pesticides for fish [Translation from: Izv.gos.nauchno-issled.Inst.ozern.rybn.Khoz. 121, 95-96, 1977]

In the piscicide laboratory of GosNIORKh over a series of years was carried out the ichthyological evaluation of different agricultural pesticides, used both in our country and abroad. In all more than 300 different chemical substances were tested. Here, it was established that around 10% of them possessed high ichthyotoxic properties. Experiments were conducted under laboratory conditions on aquarium fish in groups, and also on representatives of different species of lake ichthyofauna. The basic criterion of evaluation of toxicity was the death of experimental fish during 120 hours. This short paper summarises the findings of this reasearch and offers a table presenting acute toxicity of pesticides for fish.

The ecological impact of different mechanisms of chronic sub-lethal toxicity on feeding and respiratory physiology

Sub-lethal toxicity tests, such as the scope-for-growth test, reveal simple relationships between measures of contaminant concentration and effect on respiratory and feeding physiology. Simple models are presented to investigate the potential impact of different mechanisms of chronic sub-lethal toxicity on these physiological processes. Since environmental quality is variable, even in unimpacted environments, toxicants may have differentially greater impacts in poor compared to higher quality environments. The models illustrate the implications of different degrees and mechanisms of toxicity in response to variability in the quality of the feeding environment, and variability in standard metabolic rate. The models suggest that the relationships between measured degrees of toxic stress, and the maintenance ration required to maintain zero scope-for-growth, may be highly nonlinear. In addition it may be possible to define critical levels of sub-lethal toxic effect above which no environment is of sufficient quality to permit prolonged survival.

A Delphi Process to Optimize Quality and Performance of Drug Evaluation in Neonates

Background: Neonatal trials remain difficult to conduct for several reasons: in particular the need for study sites to have an existing infrastructure in place, with trained investigators and validated quality procedures to ensure good clinical, laboratory practices and a respect for high ethical standards. The objective of this work was to identify the major criteria considered necessary for selecting neonatal intensive care units that are able to perform drug evaluations competently. Methodology and Main Findings: This Delphi process was conducted with an international multidisciplinary panel of 25 experts from 13 countries, selected to be part of two committees (a scientific committee and an expert committee), in order to validate criteria required to perform drug evaluation in neonates. Eighty six items were initially selected and classified under 7 headings: "NICUs description - Level of care'' (21), "Ability to perform drug trials: NICU organization and processes (15), "Research Experience'' (12), "Scientific competencies and area of expertise'' (8), "Quality Management'' (16), "Training and educational capacity'' (8) and "Public involvement'' (6). Sixty-one items were retained and headings were rearranged after the first round, 34 were selected after the second round. A third round was required to validate 13 additional items. The final set includes 47 items divided under 5 headings. Conclusion: A set of 47 relevant criteria will help to NICUs that want to implement, conduct or participate in drug trials within a neonatal network identify important issues to be aware of. Summary Points: 1) Neonatal trials remain difficult to conduct for several reasons: in particular the need for study sites to have an existing infrastructure in place, with trained investigators and validated quality procedures to ensure good clinical, laboratory practices and a respect for high ethical standards. 2) The present Delphi study was conducted with an international multidisciplinary panel of 25 experts from 13 countries and aims to identify the major criteria considered necessary for selecting neonatal intensive care units (NICUs) that are able to perform drug evaluations competently. 3) Of the 86 items initially selected and classified under 7 headings - "NICUs description - Level of care'' (21), "Ability to perform drug trials: NICU organization and processes (15), "Research Experience'' (12), "Scientific competencies and area of expertise'' (8), "Quality Management'' (16), "Training and educational capacity'' (8) and "Public involvement'' (6) - 47 items were selected following a three rounds Delphi process. 4) The present consensus will help NICUs to implement, conduct or participate in drug trials within a neonatal network.

Biofunctionalization of alginate microcapsules: advances in cell-based drug delivery systems

172 p.

Modular Multimodal Iron Oxide-Based Nanocarriers for Image-Guided dsRNA Immunostimulation and Platinum Anticancer Drug Design

237 p.

Evaluation of marine phytoplankton toxicity by application of marine invertebrate bioassays

The dinoflagellate Alexandrium minutum and the haptophyte Prymnesium parvum are well known for their toxin production and negative effects in marine coastal environments. A. minutum produces toxins which cause paralytic shellfish poisoning in humans and can affect copepods, shellfish and other marine organisms. Toxins of P. parvum are associated with massive fish mortalities resulting in negative impacts on the marine ecosystem and large economic losses in commercial aquaculture. The aim of this work is to improve our knowledge about the reliability of the use of marine invertebrate bioassays to detect microalgae toxicity, by performing: (i) a 24- to 48-h test with the brine shrimp Artemia franciscana; (ii) a 48-hour embryo-larval toxicity test with the sea urchin Paracentrotus lividus; and (iii) a 72-h test with the amphipod Corophium multisetosum. The results indicate that A. franciscana and P. lividus larvae are sensitive to the toxicity of A. minutum and P. parvum. LC50 comparison analysis between the tested organisms reveals that A. franciscana is the most sensitive organism for A. minutum. These findings suggest that the use of different organizational biological level bioassays appears to be a suitable tool for A. minutum and P. parvum toxicity assessment.

Exploring new labelling strategies for boronated compounds: towards fast development and efficient assessment of BNCT drug candidates

208 p.

Pharmacogenetics of MicroRNAs and MicroRNAs Biogenesis Machinery in Pediatric Acute Lymphoblastic Leukemia

Despite the clinical success of acute lymphoblastic leukemia (ALL) therapy, toxicity is frequent. Therefore, it would be useful to identify predictors of adverse effects. In the last years, several studies have investigated the relationship between genetic variation and treatment-related toxicity. However, most of these studies are focused in coding regions. Nowadays, it is known that regions that do not codify proteins, such as microRNAs (miRNAs), may have an important regulatory function. MiRNAs can regulate the expression of genes affecting drug response. In fact, the expression of some of those miRNAs has been associated with drug response. Genetic variations affecting miRNAs can modify their function, which may lead to drug sensitivity. The aim of this study was to detect new toxicity markers in pediatric B-ALL, studying miRNA-related polymorphisms, which can affect miRNA levels and function. We analyzed 118 SNPs in pre-miRNAs and miRNA processing genes in association with toxicity in 152 pediatric B-ALL patients all treated with the same protocol (LAL/SHOP). Among the results found, we detected for the first time an association between rs639174 in DROSHA and vomits that remained statistically significant after FDR correction. DROSHA had been associated with alterations in miRNAs expression, which could affect genes involved in drug transport. This suggests that miRNA-related SNPs could be a useful tool for toxicity prediction in pediatric B-ALL.

Avaliação da toxicidade da emodina e sua associação com radiação ultravioleta A em cepas de Escherichia coli e células da linhagem A549

A emodina é uma antraquinona estruturalmente semelhante à aloe-emodina e ambas tem sido apontadas como capazes de causar lesões oxidativas pela produção de ERO. Sua presença em produtos dermocosméticos e de higiene pessoal, associada às informações de que a fotoativação de antraquinonas levaria ao aumento de lesões oxidativas causadas por ERO, torna relevante o estudo da associação da emodina com a radiação UVA. O objetivo desse trabalho foi avaliar a citotoxicidade induzida pela associação da emodina com doses subletais de radiação UVA, em células de Escherichia coli (selvagem e cepas deficientes em enzimas do BER), através de ensaios de sobrevivência bacteriana (taxa de dose de UVA igual a 20J/m/s, totalizando 108kJ/m ao final de 90min de experimento), e em células da linhagem A549 pela exclusão do corante azul de tripan e sobrevivência clonogênica(taxa de dose de UVA igual a 20J/m/s, totalizando 36kJ/m ao final de 30min de experimento). Além disso, a genotoxicidade desses agentes foi estudada por eletroforese em gel de agarose de DNA plasmidial (taxa de dose de UVA igual a 16J/m/s, totalizando 57,6kJ/m ao final de 60min de experimento). De acordo com os resultados: i) Concentrações iguais ou abaixo de 5,55mM de emodina não alteraram a sobrevivência em nenhuma das cepas estudas; ii) As proteínas Xth e Fpg parecem ter um papel importante no reparo das lesões causadas pela emodina, em altas concentrações, sugerindo a participação do reparo por excisão de bases (BER) nesse processo; iii) A associação da emodina com a radiação UVA se mostrou citotóxica em todas as cepas de E. coli; iv) O gene nfo foi o mais importante na resistência bacteriana às lesões induzidas pela associação dos dois agentes, reforçando o envolvimento do BER e indicando uma possível participação do reparo por incisão de nucleotídeos (NIR); v) A emodina parece ter interagido com o DNA plasmidial, alterando seu padrão de migração no gel de agarose; vi) Em células da linhagem A549, a emodina causa efeitos tóxicos imediatos que parecem ser reparados ao longo do tempo. Porém, quando a droga permaneceu por 24 horas em contato com as células, houve uma diminuição na sobrevivência celular que parece ser dosedependente; vii) As concentrações de 10μM e 25μM de emodina, quando associadas ao UVA, se mostraram responsáveis pela redução de mais de 50% na sobrevivência nas células A549, chegando a 100% de morte quando a concentração de emodina foi de 50μM; viii) A radiação UVA potencializou os efeitos citotóxicos da emodina, nos 2 modelos experimentais do presente estudo, indicando que a interação da emodina com a radiação UVA seja genotóxica e portanto prejudicial à saúde.

Mechanisms of Toxicity of Ag Nanoparticles in Comparison to Bulk and Ionic Ag on Mussel Hemocytes and Gill Cells

Silver nanoparticles (Ag NPs) are increasingly used in many products and are expected to end up in the aquatic environment. Mussels have been proposed as marine model species to evaluate NP toxicity in vitro. The objective of this work was to assess the mechanisms of toxicity of Ag NPs on mussel hemocytes and gill cells, in comparison to ionic and bulk Ag. Firstly, cytotoxicity of commercial and maltose stabilized Ag NPs was screened in parallel with the ionic and bulk forms at a wide range of concentrations in isolated mussel cells using cell viability assays. Toxicity of maltose alone was also tested. LC50 values were calculated and the most toxic Ag NPs tested were selected for a second step where sublethal concentrations of each Ag form were tested using a wide array of mechanistic tests in both cell types. Maltose-stabilized Ag NPs showed size-dependent cytotoxicity, smaller (20 nm) NPs being more toxic than larger (40 and 100 nm) NPs. Maltose alone provoked minor effects on cell viability. Ionic Ag was the most cytotoxic Ag form tested whereas bulk Ag showed similar cytotoxicity to the commercial Ag NPs. Main mechanisms of action of Ag NPs involved oxidative stress and genotoxicity in the two cell types, activation of lysosomal AcP activity, disruption of actin cytoskeleton and stimulation of phagocytosis in hemocytes and increase of MXR transport activity and inhibition of Na-K-ATPase in gill cells. Similar effects were observed after exposure to ionic and bulk Ag in the two cell types, although generally effects were more marked for the ionic form. In conclusion, results suggest that most observed responses were due at least in part to dissolved Ag.

Ação do Glucantime sobre macrófagos de camundongos

Os antimoniais pentavalentes, tais como o Glucantime, são geralmente usados como fármacos de primeira escolha para o tratamento das leishmanioses, no entanto seu mecanismo de ação não é completamente esclarecido. Atua contra formas amastigotas intracelulares de Leishmania sp, comprometendo o potencial redox levando danos ao DNA do parasito. Alguns trabalhos sugerem que o Glucantime aumenta a capacidade fagocítica e a produção de TNF-alfa por fagócitos. O objetivo deste estudo foi avaliar a capacidade do Glucantime modular a atividade do macrófago, a principal célula hospedeira da Leishmania. Inicialmente, a toxicidade do Glucantime foi testada sobre macrófagos peritoneais de camundongos BALB/c, tratando as monocamadas in vitro por 48 horas. A viabilidade celular foi avaliada pelo método do MTT. A capacidade do Glucantime (0,1, 1 e 10 mg/ml) modular os macrófagos foi avaliada tratando as monocamadas de macrófagos peritoneais por 24 horas antes da infecção com Leishmania braziliensis. Após 48 horas de incubação com meio de cultura foi avaliado o índice de infecção por contagem. Antes e após a infecção foram analisados a produção de óxido nítrico (NO) pelo método de Griess, espécies reativas de oxigênio (EROS) por fluorimetria usando a sonda H2DCFDA e a produção de citocinas por ELISA. Para avaliar se o Glucantime seria capaz de modular macrófagos in vivo, camundongos suíços foram tratados por 5 dias consecutivos com 8 mg de Glucantime pela via intraperitoneal. Macrófagos peritôneais foram avaliados quanto a sua capacidade de controlar a infecção in vitro com L. braziliensis. Os resultados mostraram que nas concentrações até 10 mg/ml, o Glucantime não alterou a viabilidade dos macrófagos in vitro. O pré-tratamento dos macrófagos com Glucantime nas concentrações de 0.1mg/mL, 1mg/mL e 10mg/mL, foi capaz de reduzir o índice de infecção em 49%, 74% e 85%, respectivamente. Em macrófagos não infectados a produção de NO foi aumentada na concentração de 10mg/ml de Glucantime. O tratamento com 1 e 10 mg/ml de Glucantime foi capaz de aumentar significativamente a produção de EROs (p<0,05 e p<0.01, respectivamente) e a produção IL-12 (p<0,05), mas a IL-10 não foi alterada. Não houve alterações significativas desses parâmetros em relação ao controle após a infecção com L. braziliensis. Os macrófagos oriundos dos animais tratados com Glucantime foram capazes de reduzir o índice de infecção por L. braziliensis (p<0,05). Esses resultados sugerem que o Glucantime é capaz de ativar os macrófagos e esse efeito pode contribuir para o mecanismo de ação desse fármaco.