Biological and biochemical properties of the Brazilian Potamotrygon stingrays : Potamotrygon cf. scobina and Potamotrygon gr. orbignyi

Stingrays of the family Potamotrygonidae are widespread throughout river systems of South America that drain into the Atlantic Ocean. Some species are endemic to the most extreme freshwater environment of the Brazil and cause frequent accidents to humans. The envenomation causes immediate, local, and intense pain, soft tissue edema, and a variable extent of bleeding. The present study was carried out in order to describe the principal biological and some biochemical properties of the Brazilian Potamotrygon fish venoms (Potamotrygon cf. scobina and P. gr. orbignyi). Both stingray venoms induced significant edematogenic and nociceptive responses in mice. Edematogenic and nociceptive responses were reduced when the venom was incubated at 37 or 56 degrees C. The results showed striking augments of leukocytes rolling and adherent cells to the endothelium of cremaster mice induced by both venoms. The data also presented that injection of both venoms induced necrosis, low level of proteolytic activity, without inducing haemorrhage. But when the venoms of both stingray species were injected together with their mucus secretion, the necrotizing activity was more vigorous. The present study provided in vivo evidence of toxic effects for P. cf. scobina and P. gr. orbignyi venoms. (c) 2006 Elsevier Ltd. All fights reserved.

Biological and Biochemical Characterization of Mutants of the RNase II family of enzymes

Dissertação para a obtenção de grau de doutor em Biologia pelo Instituto de Tecnologia Química e Biológica. Universidade Nova de Lisboa.

BIOLOGICAL AND BIOCHEMICAL CHARACTERIZATION OF MURINE-TUMOR SPECIFIC TRANSPLANTATION ANTIGENS (ISOTACHOPHORESIS, IMMUNOGENICITY)

Tumor-specific transplantation antigens (TSTA) are individually distinct neoantigens expressed on the cells of chemically-induced neoplasms. TSTA are operationally defined by immunization of syngeneic mice against challenge with viable tumor cells. Immunization with cell surface or extracted TSTA induces specific resistance to transplanted tumor cells. The biological and biochemical nature of TSTA was investigated in the 3-methylcholanthrene-induced fibrosarcomas of female C3H/HeJ mice, MCA-F and MCA-D. Tumor cell suspensions were extracted by treatment with 3M KCl or 2.5% butanol solutions and the TSTA was partially purified by preparative isoelectric focusing. The isoelectric pH of TSTA purified from 3M KCl extracts was 5.8-6.0, and from butanol extracts was 6.4-6.6. Whereas immunization with 10('5) and 10('6) irradiated tumor cells induces complete rejection of tumor cell challenge over a two-fold-log dose range, immunization with ug quantities within a one-fold-log dose range of extracted TSTA induces only partial resistance to tumor challenge. Reduced immunogenicity of extracted TSTA is hypothesized to result from immunization of mice with insufficiently purified TSTA preparations. The hypothesis predicts that immunization with highly purified TSTA, free from interfering substances, induces complete rejection of tumor challenge over a broad dose range. To test the hypothesis preparative isotachophoresis (pITP) was used to purify TSTA from electrofocused TSTA fractions. Significant purification was achieved, as immunization with 15 pg to 1.5 ug (5 logs) of pITP-purified TSTA extracted from the MCA-F, or with 1 pg to 10 ng (4 logs) of TSTA from the MCA-D tumor induced specific resistance to tumor challenge. Despite 50,000 fold purification of TSTA, immunization induced partial, not complete, rejection of transplanted tumor cells. This suggests a clear dissociation of the immunogenicity and purification of extracted TSTA, indicating that the induction of partial immunity to tumor challenge is an intrinsic property of extracted TSTA.^

Certain biological and biochemical aspects of the use of ionizing radiation in immunization against parasitic diseases, Yugoslavia.

Translated from Spomenica u cast novoizabranih clanova Sprske akademije nauka i umetnosti. Serbo-Croation, 1967, pp. 113-121.

A review of Australian classical biological control of weeds programs and research activities over the past 12 years

Considerable progress has been made towards the successful classical biological control of many of Australia’s exotic weeds over the past decade. Some 43 new arthropod or pathogen agents were released in 19 projects. Effective biological control was achieved in several projects with the outstanding successes being the control of rubber vine, Cryptostegia grandiflora, and bridal creeper, Asparagus asparagoides. Significant developments also occurred in target prioritization, procedures for target and agent approval, funding, infrastructure and cooperation between agencies. Scientific developments included greater emphasis on climate matching, plant and agent phylogeny, molecular diagnostics, agent prioritization and agent evaluation.

Mutations in the nucleotide binding and hydrolysis domains of Helicobacter pylori MutS2 lead to altered biochemical activities and inactivation of its in vivo function

Background: Helicobacter pylori MutS2 (HpMutS2), an inhibitor of recombination during transformation is a non-specific nuclease with two catalytic sites, both of which are essential for its anti-recombinase activity. Although HpMutS2 belongs to a highly conserved family of ABC transporter ATPases, the role of its ATP binding and hydrolysis activities remains elusive. Results: To explore the putative role of ATP binding and hydrolysis activities of HpMutS2 we specifically generated point mutations in the nucleotide-binding Walker-A (HpMutS2-G338R) and hydrolysis Walker-B (HpMutS2-E413A) domains of the protein. Compared to wild-type protein, HpMutS2-G338R exhibited similar to 2.5-fold lower affinity for both ATP and ADP while ATP hydrolysis was reduced by similar to 3-fold. Nucleotide binding efficiencies of HpMutS2-E413A were not significantly altered; however the ATP hydrolysis was reduced by similar to 10-fold. Although mutations in the Walker-A and Walker-B motifs of HpMutS2 only partially reduced its ability to bind and hydrolyze ATP, we demonstrate that these mutants not only exhibited alterations in the conformation, DNA binding and nuclease activities of the protein but failed to complement the hyper-recombinant phenotype displayed by mutS2-disrupted strain of H. pylori. In addition, we show that the nucleotide cofactor modulates the conformation, DNA binding and nuclease activities of HpMutS2. Conclusions: These data describe a strong crosstalk between the ATPase, DNA binding, and nuclease activities of HpMutS2. Furthermore these data show that both, ATP binding and hydrolysis activities of HpMutS2 are essential for the in vivo anti-recombinase function of the protein.

EFFECTS OF IRON ON PHYSIOLOGICAL AND BIOCHEMICAL CHARACTERISTICS OF Microcystis wesenbergii (Kom.) Kom. (Cyanobacterium)

Iron is an essential trace element for biological requirements of phytoplankton. Effects of iron on physiological and biochemical characteristics of Microcystis wesenbergii were conducted in this study. Results showed that 0.01 mu M [Fe3+] seriously inhibited growth and chlorophyll synthesis of M. wesenbergii, and induced temporary increase of ATPase activities, however, NR. ACP and ALP activities were restrained by iron limitation. Interestingly, iron addition on day 8 resulted in the gradual restoration of structures and functions of above enzymes and resisted a variety of stresses from iron limitation. M. wesenbergii in 10 mu M [Fe3+] treatment group grew normally. enzymes maintained normal levels, and residual phosphate contents in cultures first sharply decreased, then smoothly as M. wesenbergii has a characteristic of luxury consumption of phosphorus. Above parameters in 100 mu M [Fe3+] treatment group were almost same with those in 10 mu M [Fe3+] treatment group except for NR, ACP and ALP activities. In 100 mu M [Fe3+] treatment group, activities of ACP and ALP had temporary increase because phosphate and ferric iron could form insoluble compound - ferric phosphate (Fe3PO4) through adsorption effect. resulting in lack of bioavailable phosphate in culture media. The experiment suggested that too low or too high iron can affect obviously physiological and biochemical characteristics of M. wesenbergii.

Synthesis and biological activities of polyphenolic hybrids - evaluation of anti-cancer, anti-inflammatory and antioxidant activities

Os compostos polifenólicos constituem uma classe de metabolitos secundários de plantas, mas existe também uma enorme quantidade de derivados sintéticos ou semi-sintéticos contendo múltiplas unidades fenólicas. Estes compostos apresentam importantes características biológicas, que dependem das suas estruturas básicas. Certos derivados desta família de compostos, tais como flavonoides, cromonas e cumarinas contribuem para os benefícios da dieta humana, e partilham o núcleo de benzopiran-(2 e 4)-ona ou benzofuran-3-ona. A presente dissertação inclui uma introdução geral e três capítulos que descrevem as novas rotas sintéticas estabelecidas para a preparação de novos híbridos de diversos compostos polifenólicos, assim como a sua elucidação estrutural e termina com a presentação dos resultados da avaliação biológica desses mesmos compostos. No segundo capítulo discute-se a preparação de híbridos de pirimidina- e imidazolidina-polifenóis, especialmente a síntese diastereoseletiva de novos híbridos benzofuran-3-ona-hidantoína e derivados de uracilo. A rota sintética envolve a ação de carbodiimidas sobre os ácidos cromona-(2- e 3)-carboxílicos num só passo ou em dois passos sequenciais, catalisada por uma base orgânica ou inorgânica. O terceiro capítulo descreve reações do tipo adições conjugadas 1,4 - hetero-ciclisações em cascata de compostos 1,3-dicarbonílicos em ácido cromona-3-carboxílico catalisadas por uma base orgânica, que originaram novas cromonas, cromanonas e flavonas polissubstituídas. As bispiranonas [bispiran-2 e 4)-onas] foram elaboradas numa reacção de acoplamento da 4-hidroxicumarina ou da lactona do ácido triacético com o ácido cromona-3-carboxílico ou precursores formil-funcionalizados (ω-formil-2’-hydroxy acetofenonas e cromona-3-carbaldeídos) utilizando organocatálise básica. Finalmente, alargou-se o estudo das adições conjugadas 1,4 para uma variedade de 4-hidroxipiran-2-onas e cetonas α,β-insaturadas para originar novos análogos de warfarina. Obteve-se uma variedade de estruturas complexas por hibridação das unidades de 4-hidroxicumarina ou da lactona do ácido triacético com os novos derivados de cromonas polissubstituídas. Todos as reações foram executadas em condições suaves e ambientalmente favoráveis, utilizando a 4-pirrolidinopiridina como organocatalisador básico. As estruturas dos novos híbridos polifenólicos foram caracterizados por técnicas espectroscópicas de alta resolução, incluindo espectroscopia de ressonância magnética nuclear (1D e 2D) e por difractometria de raios-X, que nos permitiram resolver o complexidade estrutural dos compostos sintetizados. O quarto capítulo apresenta os resultados da avaliação biológica obtidos com os híbridos polifenólicos sintetizados neste trabalho, mostrando a possibilidade de seu envolvimento na terapia do cancro. A maioria dos compostos foram avaliados quanto ao seu efeito sobre a citotoxicidade e proliferação de células leucémicas e ao seu envolvimento na regulação de via pró-inflamatória NF-kB, na qual, os híbridos de biscumarinas exibiram actividades elevadas (IC50 = 6-19 μM para inibição de NF-kB depois de 8 horas de incubação e IC50 = 15-39 μM para efeitos citotóxicos em células cancerosas, após 24 horas de incubação). Uma inibição moderada das enzimas HDAC e Cdc25 foi induzida pelos derivados de benzofuran-3-ona-hidantoína. Catorze dos novos derivados polifenólicos polissubstituídos, tendo como estrutura básica a benzopiran-4-ona, foram avaliados pela sua actividade quimiopreventiva do cancro mediada pela indução de sinalização citoprotectora Nrf2 (fator 2 relacionado com o fator nuclear da proteína E2) e capacidade para inibir a proliferação das células de cancro da mama. Os derivados da classe das cromanonas foram identificados como os indutores mais potentes da actividade Nrf2. As concentrações necessárias para aumentar a actividade de luciferase em 10 vezes (C10) foram de 2,8-21,3 μM. Todos os novos híbridos polifenólicos que apresentam atividade citotóxica e anti-proliferativa não afectam o crescimento de células saudáveis periféricas do sangue (PBMC) (IC50 > 50 μM), indicando a sua seletividade para as células cancerosas e sugerindo que alguns deles são estruturalmente interessantes para posteriores análises. A avaliação da atividade antioxidante utilizando os testes do radical livre DPPH e o poder redutor do ião férrico FRAP foram realizados em algumas estruturas híbridas polifenólicas.

Chemical and Biochemical Studies on Natural Antioxidants from sesamum Species

In the present study on natural antioxidants, the focus has been kept mainly on oil seeds, especially sesame and its by-products. Sesame, which has been under cultivation in India for centuries is called the 'Queen of oil seed crops' because of the high yield of oil obtained and the nutritional qualities of the seed, oil, and meal. Though India is the largest producer of sesame in the world, research on the various health benefits of sesame has been carried out by Japanese Sesame has an important place in the foods and tradit..ional medicine of India from time immemorial. Foreseeing the potential of sesame and its byproducts as an important antioxidant source and its availability in bulk, the present study was focussed on Sesamum species. There are not many reports on the wild species of Sesamum in India, especially of the Kerala region. Hence, in the present study we also included antioxidants of Sesamurnrnalabaricumdistributed throughout the coastal region.The important characteristics of sesame are attributed to the presence of the umquc compounds lignans. Lignans arc a group of natural products of phenyl propanoid ongm, whieh are widely distributed in nature. They display important physiological functions in plants, in human nutrition and medicine, given their extensive health promotive and curative properties. Much interest has been focussed on their effectiveness as antineoplastic agents and research in this area has revealed several modes of action by which they can regulate the growth of mammalian cells. Sesame is an important source of furofuran lignans, of which sesamin and the rare oxygenated derivative sesamoIin are the most abundant. Others include sesamol and glucosides of lignans. Sesarnin and episesamin are reported to have hypocholesterolemic effect, suppressive effect on chemically induced cancer, alleviation of allergy symptoms etc. Sesamol, sesamolin and the lignan glycosides are reported to inhibit lipid peroxidation. Present investigation on sesame and its byproducts have been carried out to explore the possibility of developing a natural antioxidant extract from available resources to be used as a substitute to synthetic ones in vegetable oils and foods. Preliminary analysis showed that sesame cake, a byproduct could still be utilized as a major source of lignans. Sesame cake, which is now used only as a cattlefeed, can be better utilized in the form of a valuable antioxidant source. The present study explains the development of a feasible process for the extraction of antioxidant compounds from sesame cake. The antioxidant extract so prepared from sesame cake has been tested for vegetable oil protection and is found to be effective at low concentration. In addition, studies also include the antioxidant, radical scavenging, anticancer, mosquitocidal and pesticidal activities of extract and individual compounds.

Microbial and biochemical soil quality indicators and their potential for differentiating areas under contrasting agricultural management regimes

The aim of this study was to examine interrelationships between functional biochemical and microbial indicators of soil quality, and their suitability to differentiate areas under contrasting agricultural management regimes. The study included five 0.8 ha areas on a sandy-loam soil which had received contrasting fertility and cropping regimes over a 5 year period. These were organically managed vegetable, vegetable -cereal and arable rotations, an organically managed grass clover ley, and a conventional cereal rotation. The organic areas had been converted from conventional cereal production 5 years prior to the start of the study. All of the biochemical analyses, including light fraction organic matter (LFOM) C and N, labile organic N (LON), dissolved organic N and water-soluble carbohydrates showed significant differences between the areas, although the nature of the relationships between the areas varied between the different parameters, and were not related to differences in total soil organic matter content. The clearest differences were seen in LFOM C and N and LON, which were higher in the organic arable area relative to the other areas. In the case of the biological parameters, there were differences between the areas for biomass-N, ATP, chitin content, and the ratios of ATP: biomass and basal respiration: biomass. For these parameters, the precise relationships between the areas varied. However, relative to the conventionally managed area, areas under organic management generally had lower biomass-N and higher ATP contents. Arbuscular mycorrhizal fungus colonization potential was extremely low in the conventional area relative to the organic areas. Further, metabolic diversity and microbial community level physiological profiles, determined by analysis of microbial community metabolism using Biolog GN plates and the activities of eight key nutrient cycling enzymes, grouped the organic areas together, but separated them from the conventional area. We conclude that microbial parameters are more effective and consistent indicators of management induced changes to soil quality than biochemical parameters, and that a variety of biochemical and microbial analyses should be used when considering the impact of management on soil quality. (C) 2004 Elsevier Ltd. All rights reserved.

CHEMICAL AND BIOCHEMICAL PROPERTIES OF ARAUCARIA ANGUSTIFOLIA (BERT.) KTZE. FOREST SOILS IN THE STATE OF SAO PAULO

Araucaria angustifolia, commonly named Araucaria, is a Brazilian native species that is intensively exploited due to its timber quality. Therefore, Araucaria is on the list of species threatened by extinction. Despite the importance of soil for forest production, little is known about the soil properties of the highly fragmented Araucaria forests. This study was designed to investigate the use of chemical and biological properties as indicators of conservation and anthropogenic disturbance of Araucaria forests in different sampling periods. The research was carried out in two State parks of Sao Paulo: Parque Estadual Turistico do Alto do Ribeira and Parque Estadual de Campos de Jordao. The biochemical properties carbon and nitrogen in microbial biomass (MB-C and MB-N), basal respiration (BR), the metabolic quotient (qCO(2)) and the following enzyme activities: beta-glucosidase, urease, and fluorescein diacetate hydrolysis (FDA) were evaluated. The sampling period (dry or rainy season) influenced the results of mainly MB-C, MB-N, BR, and qCO(2). The chemical and biochemical properties, except K content, were sensitive indicators of differences in the conservation and anthropogenic disturbance stages of Araucaria forests. Although these forests differ in biochemical and chemical properties, they are efficient in energy use and conservation, which is shown by their low qCO(2), suggesting an advanced stage of succession.

Chemical and biochemical properties of Araucaria angustifolia (Bert.) Ktze. forest soils in the state of São Paulo

Araucaria angustifolia, commonly named Araucaria, is a Brazilian native species that is intensively exploited due to its timber quality. Therefore, Araucaria is on the list of species threatened by extinction. Despite the importance of soil for forest production, little is known about the soil properties of the highly fragmented Araucaria forests. This study was designed to investigate the use of chemical and biological properties as indicators of conservation and anthropogenic disturbance of Araucaria forests in different sampling periods. The research was carried out in two State parks of São Paulo: Parque Estadual Turístico do Alto do Ribeira and Parque Estadual de Campos de Jordão. The biochemical properties carbon and nitrogen in microbial biomass (MB-C and MB-N), basal respiration (BR), the metabolic quotient (qCO2) and the following enzyme activities: β-glucosidase, urease, and fluorescein diacetate hydrolysis (FDA) were evaluated. The sampling period (dry or rainy season) influenced the results of mainly MB-C, MB-N, BR, and qCO2. The chemical and biochemical properties, except K content, were sensitive indicators of differences in the conservation and anthropogenic disturbance stages of Araucaria forests. Although these forests differ in biochemical and chemical properties, they are efficient in energy use and conservation, which is shown by their low qCO2, suggesting an advanced stage of succession.

Physiological and biochemical characteristics of polar cod (Boreogadus saida) from Kongsfjorden

Seasonality of biomarker baseline levels were studied in polar cod (Boreogadus saida), caught in Kongsfjorden, Svalbard, in April, July, September and December, 2006-2007. Physiological parameters (condition factor, gonado- and hepato-somatic indexes, energy reserves, potential metabolic activity and antifreeze activity) in polar cod were used to interpret the seasonality of potential biomarkers. The highest levels of ethoxyresorufin-O-deethylase (EROD) activity occurred concomitantly with the highest potential metabolic activity in July due to e.g. intense feeding. During pre-spawning, EROD showed significant inhibition and gender differences. Hence, its potential use in environmental monitoring should imply gender differentiation at least during this period. Glutathione S-transferase and catalase activities were stable from April to September, but changed in December suggesting a link to low biological activity. Knowledge of the biomarker baseline levels and their seasonal trends in polar cod is essential for a trustworthy interpretation of forthcoming toxicity data and environmental monitoring in the Arctic.