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BIOCHEMICAL AND PHYSIOLOGICAL BEHAVIOR OF ANTARCTIC ORGANISMS. The aim of the present paper was to carry on a report on the scientific contribution of the project “Biochemical and physiological behavior of Antarctic organisms” towards the Brazilian Antarctic Program, a project that established in 1983-1984 the biological research laboratories at the Brazilian Antarctic Station Commander Ferraz, King George Island, South Shetlands. It is given a brief report on the Project and its researchers, as well as on the elaboration of projects for the assembly of special instruments for the measurement of biological parameters from Antarctic organisms. It is also given an information on the researches carried out or being carried out whose results have being informed in National and International Meetings and Symposia, in formal publications as well as in Master Degrees and Doctors Thesis.

The aim of this work was to develop and evaluate an indirect enzyme-linked immunosorbent assay (ELISA) and made a serological screening for specific antibodies to BoHV-4. Bovine serum samples were collected from different Brazilian states and evaluated for the presence of antibodies for BoHV-4 and BoHV-1. The serological results obtained showed that the indirect ELISA assay could be applied for the detection of specific antibodies for BoHV-4. The ELISA test allowed concluding that BoHV-4 is present in bovines in every Brazilian state from which serum samples were collected. The ELISA assay here standardized proved to be useful for the epidemiological studies and showed a positivity range from 1.8 to 66% in Brazil.

O genocídio perpetrado pelo nazismo ocupa um lugar destacado na história do século XX como paradigma da barbárie. Nessa condição, tornou-se também um expediente discursivo muito utilizado para legitimar ou estigmatizar (“nazificar”) certos agentes e causas. O presente trabalho historiciza e explora representações concorrentes das atrocidades nazistas, analisando como se constitui em capital simbólico mobilizado para justificar ou condenar as ações do Estado israelense que, por sua vez, busca se afirmar como porta-voz de todas as vítimas do holocausto. O artigo apresenta os primeiros usos dessa memória e os principais críticos de sua manipulação discursiva. Na sequência o enfoque recai sobre uma dimensão pouco explorada da questão: as evocações do nazismo para condenar a ocupação e certas ações israelenses, dos anos 1940 até 2014. Ao final, esse uso específico é contraposto à bibliografia especializada sobre modernidade, holocausto e barbárie.

Loxosceles spiders are responsible for serious human envenomations worldwide. The collection of symptoms found in victims after accidents is called loxoscelism and is characterized by two clinical conditions: cutaneous loxoscelism and systemic loxocelism. The only specific treatment is serum therapy, in which an antiserum produced with Loxosceles venom is administered to the victims after spider accidents. Our aim was to improve our knowledge, regarding the immunological relationship among toxins from the most epidemiologic important species in Brazil (Loxosceles intermedia, Loxosceles gaucho and Loxosceles laeta). Immunoassays using spider venoms and L. intermedia recombinant toxins were performed and their cross-reactivity assessed. The biological conservation of the main Loxosceles toxins (Phospholipases-D, Astacin-like metalloproteases, Hyaluronidase, ICK-insecticide peptide and TCTP-histamine releasing factor) were investigated. An in silico analysis of the putative epitopes was performed and is discussed on the basis of the experimental results. Our data is an immunological investigation in light of biological conservation throughout the Loxosceles genus. The results bring out new insights on brown spider venom toxins for study, diagnosis and treatment of loxoscelism and putative biotechnological applications concerning immune conserved features in the toxins.

Brown spider (Loxosceles genus) venom can induce dermonecrotic lesions at the bite site and systemic manifestations including fever, vomiting, convulsions, disseminated intravascular coagulation, hemolytic anemia and acute renal failure. The venom is composed of a mixture of proteins with several molecules biochemically and biologically well characterized. The mechanism by which the venom induces renal damage is unknown. By using mice exposed to Loxosceles intermedia recombinant dermonecrotic toxin (LiRecDT), we showed direct induction of renal injuries. Microscopic analysis of renal biopsies from dermonecrotic toxin-treated mice showed histological alterations including glomerular edema and tubular necrosis. Hyalinization of tubules with deposition of proteinaceous material in the tubule lumen, tubule epithelial cell vacuoles, tubular edema and epithelial cell lysis was also observed. Leukocytic infiltration was neither observed in the glomerulus nor the tubules. Renal vessels showed no sign of inflammatory response. Additionally, biochemical analyses showed such toxin-induced changes in renal function as urine alkalinization, hematuria and azotemia with elevation of blood urea nitrogen levels. Immunofluorescence with dermonecrotic toxin antibodies and confocal microscopy analysis showed deposition and direct binding of this toxin to renal intrinsic structures. By immunoblotting with a hyperimmune dermonecrotic toxin antiserum on renal lysates from toxin-treated mice, we detected a positive signal at the region of 33-35 kDa, which strengthens the idea that renal failure is directly induced by dermonecrotic toxin. Immunofluorescence reaction with dermonecrotic toxin antibodies revealed deposition and binding of this toxin directly in MDCK epithelial cells in culture. Similarly, dermonecrotic toxin treatment caused morphological alterations of MDCK cells including cytoplasmic vacuoles, blebs, evoked impaired spreading and detached cells from each other and from culture substratum. In addition, dermonecrotic toxin treatment of MDCK cells changed their viability evaluated by XTT and Neutral-Red Uptake methodologies. The present results point to brown spider dermonecrotic toxin cytotoxicity upon renal structures in vivo and renal cells in vitro and provide experimental evidence that this brown spider toxin is directly involved in nephrotoxicity evoked during Loxosceles spider venom accidents.

Cellulose synthesis in bacteria is a complex process involving the concerted action of several enzymes whose genes are often organized in operons. This process influences many fundamental physiological aspects such as bacteria and host interaction, biofilm formation, among others. Although it might sound contradictory, the participation of cellulose-degrading enzymes is critical to this process. The presence of endoglucanases from family 8 of glycosyl hydrolases (GH8) in bacterial cellulose synthase (Bcs) complex has been described in different bacteria, including the model organism Komagataeibacter xylinus; however, their role in this process is not completely understood. In this study, we describe the biochemical characterization and three-dimensional structure of a novel GH8 member from Raoultella ornithinolytica, named AfmE1, which was previously identified by our group from the metagenomic analysis of the giant snail Achatina fulica. Our results demonstrated that AfmE1 is an endo-β-1,4-glucanase, with maximum activity in acidic to neutral pH over a wide temperature range. This enzyme cleaves cello-oligosaccharides with a degree of polymerization ≥ 5 and presents six glucosyl-binding subsites. The structural comparison of AfmE1 with other GH8 endoglucanases showed significant structural dissimilarities in the catalytic cleft, particularly in the subsite +3, which correlate with different functional mechanisms, such as the recognition of substrate molecules having different arrangements and crystallinities. Together, these findings provide new insights into molecular and structural features of evolutionarily conserved endoglucanases from the bacterial cellulose biosynthetic machinery.

Genetic parameters for milk yield, fat and protein milk content and "mozzarella" index were estimated in the population of Romanian Buffalo from șercaia Research and Development Station. A total 609 milk yield and associated characters records, belonging to 87 females, which coming from 11 sire families, for 7 lactations were analyzed. The method used for genetic parameters estimates was REML. There was a large variability in all analyzed traits. Estimated heritability was small to medium for al examined traits. The milk yield is negatively correlated with fat and protein content, and weak positive with "mozzarella" index. Fat and protein milk content are high positive correlated with mozzarella production. In the long term, the female selection for milk yield will be (make) to the detriment of quality. Genetic improvement of the quantity of mozzarella will be make only on account of milk constituents that impart its quality. According to heritability values, in the process of female genetic evaluation appears advisable to use additional information sources in order to increase the accuracy of selection.

The present work aimed to analyze the ffectiveness of the principle of human dignity in confrontation with Article 13 of the Brazilian Civil Code, specifically regarding to the concretion, by the law sistem, of individual freedom and anthropological conception of man with moral autonomy, these two logical consequences of the mentioned constitutional pillar. To that end, we introduced the topic to indicate that strictly moral conceptions can not legitimize the analysis within a democratic state. We also evaluated the principle of human dignity as a constitutional option for a notion of an individual free and morally autonomous, and then we compared this with the premise Personality Rights. Finally, starting from the initial basis and taking into account the theory of the minimum ethical, it was analyzed the article 13 of the Brazilian Civil Code.

Lâmpadas fluorescentes (LF) contêm mercúrio em sua composição, um metal pesado que pode causar danos à saúde e ao meio ambiente. As LF são amplamente consumidas no Brasil, porém o descarte correto de LF usadas não é amplamente divulgado e os índices de reciclagem são baixos. O objetivo deste estudo foi demonstrar a relevância do descarte adequado, avaliar as tecnologias de reciclagem e a destinação dos materiais, por meio de estudo comparativo entre Brasil e Alemanha. Através de visitas a recicladoras e análise das tecnologias adotadas nos dois países, foi verificado que no Brasil o vidro é destinado à indústria de cerâmica, enquanto que na Alemanha é destinado à fabricação de tubos para LF. Além disso, foi observado que o mercúrio apresenta diversas possibilidades de uso. Um ponto a ser explorado nos dois países, e de grande potencial para reciclagem, é a recuperação de terras raras do pó fosfórico presente nas LF. Tanto Brasil como Alemanha possuem oportunidades de melhoria na logística reversa de LF, por meio da qual se torna possível elevar os índices de reciclagem e reduzir os impactos ambientais causados pelo descarte inadequado de LF usadas. Fluorescent lamps (FL) contain mercury in their composition, a heavy metal that can be a health hazard and damage the environment. The FL are widely consumed in Brazil, although the right disposal of FL is not well known and the recycling rate is low. This study objective is to demonstrate the importance of proper disposal and to evaluate recycling technologies and material destination through a comparative study between Brazil and Germany. In visits to recycling companies in both countries and analyzing the used technologies, it was verified that in Brazil the glass is sent to ceramics industry, while in Germany it is mainly sent for the manufacture of FL tubes. Also, the mercury has several applications. A point to be explored in both countries, with great recycling potential, is the recovery of the rare earth from phosphoric powder from FL. Both Brazil and Germany have improvement opportunities in the reverse logistics of FL, through which it becomes possible to increase recycling rates and reduce the environmental impact caused by the inappropriate disposal of used FL.

As the number of prospective students decreases and competition intensifies, student satisfaction should be a core element of the universities` marketing strategy. Students` satisfaction is influenced by the degree to which higher education institutions meet or exceed their expectations. Higher education managers in general and business higher education managers in particular need to determine the students` level of satisfaction in order to assess their performance. This, however, is difficult to achieve because satisfaction is an abstract concept and no clear consensus exists over its definition and measurement. Although, on account of education`s complexity and peculiarities, the investigation of satisfaction with educational services has been laborious, the literature review reveals a growing number of papers dealing with this issue. These papers consider the factors affecting satisfaction mostly as individual coefficients and ignore the latent relationships between constructs. However, since the determinants of partial student satisfaction simultaneously manifest in the overall satisfaction, it may be considered that an individual approach to them, would partially cover the issue being studied. For this reason, the purpose of this paper is to develop a model for measuring student satisfaction with business education services, a model which should be approached holistically and whose latent structure should be taken into account. The proposed measurement tool, based on the study of the literature, was tested on the students of a business educational institution and the results confirm the goodness-of-fit, reliability and validity of the model. The developed tool encompasses a number of factors that allow the assessment of student satisfaction with a wide range of services provided by business education institutions and relate to: educational process (syllabus, training of teaching staff, examination policy), administrative staff, admission process (information, staff), management of the faculty, image of the faculty, library, general information, cafeteria, campus climate, tutors, collaboration between faculty and business environment, international cooperation, leisure activities supported by the faculty.

The isolated intestinal smooth muscle of Antarctic fishes Notothenia neglecta and the icefish Chaenocephalus aceratus displayed a sustained tonic contraction depending upon the temperature of the nutritive bath. The contraction was maximal at 2℃, relaxing proportionally with rise of the temperature, from 2℃ to 18℃. The contractile responses to acetylcholine gradually increased in proportion to the rise of temperature. By chelating calcium ions, EDTA caused a reversible relaxation of the contracted muscle at 2.0℃. This effect was eliminated by washing out the drug from the muscle nutritive bath. Similar results were found with the administration of verapamil, a calcium channel blocker. However, after this drug, tonic contraction was not restored by washout. Such contraction was partially inhibited by the muscarinic blocking drug atropine. Results show that the tonic contraction is an active response, dependent on the entry of calcium into the cell and mediated by cholinergic receptors. The temperature-related tonic contraction is not a general event in poikilotherm animals, since it did not occur in the Bufo marinus intestinal smooth muscle, in similar experiments carried out as a control, in a tropical environment. The responsiveness of the smooth muscle to acetylcholine appeared to increase with temperature, suggesting lack of adaptation of the organ to low Antarctic temperature.

アデリーペンギン属の, ゼンツーペンギンPygoscelis papua, アゴヒゲペンギンPygoscelis antarctica, アデリーペンギンPygoscelis adeliaeの肝臓・腎臓中のアルギナーゼ量と動力学についての研究を行った。ゼンツーポンギンでは24時間絶食個体に比べて, 給餌個体の血中尿素量は高かった。アゴヒゲペンギンの場合, 給餌個体の血中尿素量は, 絶食個体の1.5倍であった。そして, 給餌したゼンツーペンギンの血中尿素濃度は, 絶食個体の3.5倍であった。また, アゴヒゲペンギン給餌個体の尿酸値は絶食個体の2倍, ゼンツーペンギンは4.8倍であった。肝臓中のアルギナーゼ活性は, アゴヒゲペンギン活性は, アゴヒゲペンギン成体でタンパク質1mg当たり561mU, ゼンツーペンギン成体では208mUであった。肝臓ホモジナイズから得たアルギナーゼのL-アルギニンに対するミカエリス定数KmはpH9.5で16.0±2.0mMであった。一般に鳥類のアルギナーゼは, 100-200mMの高いKm値を示す。高タンパク餌料, 高血中尿素量は, 肝臓アルギナーゼの量とアルギナーゼの基質に対する高い親和性の結果である。, A study was carried out on the levels and the kinetics of liver and kidney arginase from Pygoscelis penguins, the gentoo Pygoscelis papua, the chinstrap, Pygoscelis antarctica, and the Adelie, Pygoscelis adeliae. Higher values of blood urea were found in the gentoo penguins in the native state when compared with specimens maintained in the fasting state for 24 hours. In the chinstrap penguin Pygoscelis antarctica the average value for blood urea was 1.5 times higher in the native state than in the fasting condition. In the native gentoo penguin P. papua the relative increase in the blood urea concentration is as high as 3.5 times in regard to the levels found in the fasting state. In regard to the blood levels of uric acid, the difference between the native state and the fasting state is 2.0 times for P. antarctica and 4.8 times for P. papua. Specific activities of arginase assayed in penguin liver were 561 mU/mg protein and 208mU/mg protein for adult P. antarctica and P. papua respectively. Kinetic studies with arginase from penguin liver homogenates showed Km values for L-arginine of 16.0±2.0mM at pH9.5. Arginase from birds possesses in general high Km values (between 100-200mM). It seems then that the high protein diet and the high levels of blood urea of penguins are a consequence of the levels of hepatic arginase and the high affinity of this enzyme toward its substrate.

本文は鳥類赤血球中のグルコース代謝における, 有機燐酸代謝産物である, イノシトール5リン酸(IP5), 2,3ジホスホグリセリン酸(2,3-DPG)のもつ意味を知ることを目的とするものである。使用したヘキソキナーゼは, セファローズ-N-アミノヘキサノイルグルコサミンを用いたアフィニティクロマトグラフィを用いて精製した。IP5のヘキソキナーゼ抑制作用は, Mg^に対して競争的であるが, グルコース, MgATP^に対しては競争的ではない。一方, ヘキソキナーゼの活性はATP^によって抑制される。赤血球中のMg^値(2.76μmol/ml)は, 赤血球標準懸濁液中のATP, IP5の総量に比べて低い。, The aim of the present study is to show the possible implication of organic phosphate metabolites=namely inositol pentaphosphate (IP5) and 2,3-bisphosphoglycerate (2,3-DPG)=in the glucose metabolism displayed by birds erythrocytes. For this purpose, a preparation of hexokinase (HK) has been purified from chicken erythrocytes (RBC) by affinity chromatography in Sepharose-N-aminohexanoyl glucosamine and used for kinetic experiments. It has been shown that IP5 is a competitive inhibitor of HK in regard to Mg^, but not in regard to glucose and MgATP^. On the other hand, this preparation of HK is also competitively inhibited by ATP^. It has been found that the total intracellular content of Mg^ (2.76μmol/ml) is lower than the total value for the content of ATP and IP5 found in a standard packed red blood cell suspension.

The cardiac responses of the Antarctic fishes Notothema neglecta and Chaenocephalus aceratus to acute changes of temperature were studied. The fishes were surgically prepared for electrocardiogram (ECG) recording and placed in a three hour experiment in an experimental chamber containing aerated marine water. The temperature was controlled from +2℃ to -1.8℃. Presence of nitrites in the fish bath was monitored. Atropine and epinephrine were administered into the abdominal cavity through a surgically inserted cannula. Blood samples were taken at initial +2.0℃ and final -1.8℃ for cholmesterase assay. Results showed that: 1. A temperature decrease from +2.0°C to～-1.8℃ significantly depressed cardiac rate, from 26.9±27 to 5.2±1.4 beats min^ in N neglecta and from 19.0 to 3.0 beats min^ in C aceratus. In one specimen of N. neglecta, a reversible cardiac arrest of 3.0mm was observed. ECG waves did not change during the experiments, except the QRS to QRS or the Q-T intervals at lower temperatures. Respiratory rate was not significantly influenced by temperature, 2. Return of temperature to +1.5/2.0℃ immediately ceased bradycardia, 3. Administration of atropme reversed bradycardia within 5 min; 4. Serum cholmesterase activity decreased at the lowest temperature; 5. Epinephrine neither influenced the cardiac rate at any temperature nor altered cardiac waves. Results demonstrate that severe bradycardia at subzero temperatures is not only an intrinsic cardiac response to cold, but an active one, due to the higher cholinergic tone, that was not observed in the resting state at +2.0℃. No adrenergic influence on heart chronotropism has been found.

Research has been carried out on the effect of temperature on the physiological properties of mitochondria isolated from liver, cardiac muscle and encephalon of the Antarctic fish Notothenia neglecta. Mitochondria were obtained in a medium containing 0.25 M mannitol, 0.075 M sucrose, 10^ M EDTA, and 10^ M TRIS, in 10^ M phosphate buffer, pH 7.4. All assays were carried out by means of a polarograph assembled with an oxygen electrode possessing a temperature sensor. Respiration rates, oxidative phosphorylation and respiratory control values were assayed in regard to the effect of temperature. All preparations of liver, cardiac muscle and encephalon mitochondria displayed a very efficient respiration mechanism at 1℃. No general pattern for the influence of temperature increase has been found in those preparations. On the other hand, encephalon mitochondria displayed an efficient system for ATP generation by oxidative phosphorylation, but sensitive to temperature increase.

RESUMOO presente estudo teve por finalidade a análise da efetivação do princípio da dignidade da pessoa humana em confronto com o artigo 13 do Código Civil Brasileiro, no que diz respeito especificamente à concretização, pelo ordenamento jurídico pátrio, da liberdade individual e concepção antropológica do homem enquanto dotado de autonomia moral, estes decorrências lógicas do pilar constitucional anteriormente mencionado. Para tanto, foi introduzido o tema de forma a indicar que concepções estritamente morais não podem legitimar a análise dentro de um Estado democrático. Avaliou-se, então, o princípio da dignidade da pessoa humana enquanto opção constitucional por uma noção de indivíduo livre e autônomo moralmente, para depois confrontar essa premissa com os Direitos da Personalidade. Finalmente, partindo da base inicial e tendo em conta a teoria do mínimo ético, foi analisado o artigo 13 do Código Civil Brasileiro.Palavras-chave: Dignidade da Pessoa Humana, liberdade individual, autonomia moral, direitos da personalidade, mínimo ético, disponibilidade sobre o próprio corpo.ABSTRACTThe present work aimed to analyze the effectiveness of the principle of human dignity in confrontation with Article 13 of the Brazilian Civil Code, specifically regarding to the concretion, by the law sistem, of individual freedom and anthropological conception of man with moral autonomy, these two logical consequences of the mentioned constitutional pillar. To that end, we introduced the topic to indicate that strictly moral conceptions can not legitimize the analysis within a democratic state. We also evaluated the principle of human dignity as a constitutional option for a notion of an individual free and morally autonomous, and then we compared this with the premise Personality Rights. Finally, starting from the initial basis and taking into account the theory of the minimum ethical, it was analyzed the article 13 of the Brazilian Civil Code.Keywords: Human Dignity, individual freedom, moral autonomy, personality rights, ethical minimum, availability on it’s own body.

Objective To systematically identify and analyse all published literature relating to the provision of undergraduate education for preparedness in ear, nose and throat (ENT) surgery, as perceived by medical students and clinicians in the UK. Design Systematic literature review. Data sources 5 major databases were searched: MEDLINE, EMBASE, ERIC, Cochrane and Web of Science. The literature search was conducted from February to April 2015. Eligibility criteria for selecting studies Primary research or studies that report on the provision of undergraduate education for preparedness in ENT, from the perspective of medical students and clinicians in the UK. The timescale of searches was limited from 1999 onwards (ie, the past 15 years). Data extraction The literature search was conducted by 2 independent reviewers. Search terms used involved the combination and variation of 5 key concepts, namely: medical student, clinician, ENT, undergraduate medical education and UK. A data extraction form was designed for and used in this study, based on guidelines provided by the UK National Health Service (NHS) Centre for Reviews and Dissemination. Textual narrative synthesis was used for data analysis. Results A total of 7 studies were included in the final review. 4 main themes were identified: confidence in managing patients, teaching delivery, student assessment and duration of rotations. A consistent finding in this review was that the majority of final year medical students and junior doctors did not feel adequately prepared to practise ENT. Important factors influencing preparedness in ENT included the duration of clinical rotations, the opportunity for hands-on learning and formal assessment. Conclusions The findings of this review suggest the need for further development of the ENT undergraduate curricula across the UK. However, there is insufficient evidence from which to draw strong conclusions; this in itself is beneficial as it highlights a gap in the existing literature and supports the need for primary research.

In this paper, a new assay based on oxygen uptake assessed by polarography was evaluated with the aim of establishing the viability of freeze-dried BCG vaccine. An oxygen electrode possessing a temperature sensor was designed for this purpose. The polarographic method used had several advantages, particularly its rapidity and use of small amounts of biological material. These advantages are ideal for quality control of BCG vaccine.

O presente trabalho discute o posicionamento da Organização para a Unidade Africana em relação a Israel e à Questão Palestina, fundamentando em suas resoluções e naquelas da Organização das Nações Unidas. Busca-se traçar a aproximação de pan-arabismo e pan-africanismo e a evolução dos debates sobre o Oriente Médio no seio da OUA e compreender o que a teria levado a se solidarizar com a Questão Palestina e a considera-la, em 1975, literalmente uma causa africana. O recorte analisado é balizado pela ocupação dos territórios árabes por Israel, que marcou uma virada nas relações da África com esse país, e pela aprovação da resolução 3379 da Assembleia Geral da ONU, em 1975, apoiada amplamente pelos membros africanos, que considerou o sionismo uma forma de racismo.

Foi procedido um estudo do metabolismo glicídico do eritrócito de ovinos (Ideal, Suffolk e Corriedale) , de bovinos (Schleswig-Holstein vermelho-e-branco e preto-e-branco) , de eqüinos (P.S.I. e Percheron) e de caninos (Dálmata, Beagle, Dog Alemão, Basset Hound, Doberman e Pastor Alemão). Foram estudados o compartilhamento da glicose sangüínea e a sua distribuição entre o plasma e o eritrócito; o fluxo glicolítico, medido em suspensões eritrócitárias incubadas a 37°C com glicose; e os níveis de enzimas do metabolismo glicolítico (Hexoquinase, H.K.; Fosfoglicose isomerase, PGI; Fosfofructoquinase, PFK; Frutose-1, 6-bisfosfato aldolase, FruP2-ALD; Gliceraldeído-3-fosfato desidrogenase, GAPDH; Fosfogliceratoquinase, PGK; Fosfopiruvato-hidratase (Enolase) ENO; Piruvatoquinase, PK; Desidrogenase do lactato, LDH; Glicose-6-fosfato desidrogenase, G-6-PDH; Fosfogliconato desidrogenase, 6-PGDH). Em base à determinação dos níveis eritrocitários de enzimas, foram estabelecidos os perfis de enzimas da glicólise para os eritrócitos de cada espécie, com um estudo comparativo das suas concentrações e também em relação aos níveis de HK, neste último caso, com a finalidade de analisar os possíveis pontos de cruzamento sobre o eixo das abcissas. Ainda, com base nos dados de níveis de enzimas eritrocitárias, foram calculados os valores das relações entre pares de enzimas que mantêm, entre si, relação do tipo produto da reação-substrato. A atividade metabólica do eritrócito foi medida quanto à sua capacidade de produzir lactato, a partir da incubação de hemolisados com glicose ou com 2,3-Bisfosfoglice-rato. Foram estabelecidos, também, os valores de hemoglobina glicosilada dos diferentes animais estudados, denominando-se a fração obtida por eluição com fosfato-cianeto, de hemoglobina de eluição rápida (HbER) e a eluida por fosfato-cianeto com força iônica maior, de hemoglobina de eluição lenta (HbEL). As frações obtidas, a partir da cromatografia em resina catiônica BioRex 70, foram submetidas à eletroforese do tipo "DISC" em gel de poliacrilamida e as mobilidades eletroforéticas das diferentes frações obtidas, determinadas. Foi ainda estabelecida correlação entre os valores de HbER e os níveis glicêmicos encontrados, medidos em sangue total

The shortage of petroleum reserves and the increase in CO(2) emissions have raised global concerns and highlighted the importance of adopting sustainable energy sources. Second-generation ethanol made from lignocellulosic materials is considered to be one of the most promising fuels for vehicles. The giant snail Achatina fulica is an agricultural pest whose biotechnological potential has been largely untested. Here, the composition of the microbial population within the crop of this invasive land snail, as well as key genes involved in various biochemical pathways, have been explored for the first time. In a high-throughput approach, 318 Mbp of 454-Titanium shotgun metagenomic sequencing data were obtained. The predominant bacterial phylum found was Proteobacteria, followed by Bacteroidetes and Firmicutes. Viruses, Fungi, and Archaea were present to lesser extents. The functional analysis reveals a variety of microbial genes that could assist the host in the degradation of recalcitrant lignocellulose, detoxification of xenobiotics, and synthesis of essential amino acids and vitamins, contributing to the adaptability and wide-ranging diet of this snail. More than 2,700 genes encoding glycoside hydrolase (GH) domains and carbohydrate-binding modules were detected. When we compared GH profiles, we found an abundance of sequences coding for oligosaccharide-degrading enzymes (36%), very similar to those from wallabies and giant pandas, as well as many novel cellulase and hemicellulase coding sequences, which points to this model as a remarkable potential source of enzymes for the biofuel industry. Furthermore, this work is a major step toward the understanding of the unique genetic profile of the land snail holobiont.

The first questions that all scientists address when approaching a new domain are related to how well studied is the field, who and what has studied. The answers help them establish their personal areas of interest and contribution. Thus in order to help the authors concerned with the luxury domain we decided to conduct a secondary marketing research and the main purpose was to identify the studies and the trends in research in the area of marketing of luxury goods and their degree of approach, before 2005. The present study is only a part of a complex research that approaches the entire evolution in luxury research field, but due to space limitation papers were separated in two: the evolution before 2005 and the evolution after 2005. Unique by its theme, this paper is intended to be ambitious by analysing all the luxury materials to which had access the authors The results confirmed the previous statement of some authors that luxury was little researched comparing to other domains.

Orientador: Prof. Dr. Klaus Fischer Co-Orientadora: Profa. Dra. Monica Kolicheski, Prof. Leandro Wiemes Dissertação (mestrado) - Universidade Federal do Paraná, Setor de Tecnologia, Programa de Mestrado profissional em Meio Ambiente Urbano e Industrial. Defesa: Curitiba:20/06/2012 Bibliografia: fls. 127-143 Resumo: O risco ambiental de contaminação de uma única lâmpada fluorescente (LF) pode ser dito como quase nulo. Entretanto, levando em consideração que no Brasil existe uma geração anual de resíduos de LF estimada em 206 milhões de unidades e que o uso vem aumentando significativamente, principalmente, devido à política de banimento das lâmpadas incandescentes, tem-se como consequência o surgimento de uma problemática ambiental relacionada à destinação pós-consumo destes produtos. Devem-se considerar os baixos índices de reciclagem atuais. Além disso, LF contém mercúrio, um metal pesado altamente tóxico. Apesar da quantidade contida em uma lâmpada apresentar-se em miligramas, caso ocorra o rompimento de milhares de unidades, ocorre contaminação do ar, do solo e da água, ocasionando impactos ambientais irreversíveis. O objetivo deste trabalho foi demonstrar a importância da destinação adequada para LF usadas, apresentando processos de reciclagem e a estruturação da logística reversa, através de estudo comparativo entre Brasil e Alemanha. A metodologia consistiu em levantamento de dados de empresas e realização de análises para determinação do mercúrio em LF novas e usadas e em filtro de carvão ativado utilizado para reter vapor de mercúrio durante a reciclagem. Este filtro, após saturação, foi classificado como resíduo classe II. O Brasil não possui uma lei estabelecendo limites de mercúrio em LF novas. Comparando com o valor estabelecido pela diretiva da União Européia, a maioria dos resultados estaria fora dos padrões. No Brasil, a obrigatoriedade da logística reversa para LF foi instituída na Política Nacional de Resíduos Sólidos em 2010. Na Alemanha, a lei data de 2005 e inclui requisitos para recicladores, metas de reciclagem, critérios para o gerenciamento do sistema de retorno e limites de mercúrio por tipo de lâmpada. Atualmente existem quatro plantas recicladoras de LF na Alemanha e quatorze no Brasil, sendo o índice de reciclagem em relação à geração de resíduos estimado em 30% e 6%, respectivamente. As tecnologias de reciclagem de LF usadas conduzem a obtenção de frações com diferentes graus de pureza, o que impacta na destinação dos materiais obtidos. O vidro tem como principal destinação no Brasil à indústria de cerâmica e na Alemanha para a fabricação de novos tubos para lâmpadas. Outro diferencial consiste no envio do pó fluorescente para a recuperação dos elementos terras raras, atividade iniciada na Alemanha. O mercúrio tem diversos destinos, devido aos custos da purificação. O modelo proposto neste trabalho para a logística reversa de LF no Brasil contempla a criação de regulamentação federal específica definindo metas para coleta, reciclagem, critérios para o sistema de gestão, transporte, rotulagem ambiental, limites para o uso de mercúrio em lâmpadas e controle das destinações dos materiais. A estrutura do sistema constitui-se de uma associação para o gerenciamento da coleta, transporte e reciclagem, visando reduzir custos e a complexidade do sistema, bem como a fundação de entidade para registro das quantidades produzidas, importadas, coletadas e recicladas. Através da implantação da logística reversa com base nos fatores apresentados, possibilitaria elevar os índices de reciclagem, reduzir os impactos ambientais e promover a geração de novos negócios. Abstract: The environmental risk of contamination from a single fluorescent lamp (FL) can be irrelevant. However, taking into account that in Brazil there is an estimated waste generation per year of 206 million units of FL and that the use has increased significantly, mainly due to the policy of banning incandescent bulbs, has as consequence the appearance of an environmental problem related to the post-consumer destination of these products. The low indexes of current recycling must be considered. Furthermore, FL contain mercury, a highly toxic heavy metal. Although the amount in a lamp is in milligrams, in the event of breaking of thousands of units, it happens the contamination of air, soil and water, causing irreversible environmental impacts. This study aimed to demonstrate the importance of proper disposal for used FL, presenting the recycling processes and the structure of reverse logistics, through a comparative study between Brazil and Germany. The methodology was based on data collection from companies and mercury determination analyzes in used and new FL and in an activated carbon filter which is used to retain mercury vapor during recycling. This filter, after saturation, was classified as class II waste. Brazil does not have a law defining the mercury limits in new FL. Most of the results would be out of standards when compared with the value set by the EU directive. In Brazil, in 2010, the requirement of reverse logistics for FL was established in the Solid Waste National Policy. In Germany, the law dates from 2005 and it includes requirements for recyclers, recycling targets, the criteria for the return system management and mercury limits per lamp. Currently, there are four FL recycling plants in Germany and fourteen in Brazil. The recycling rate in relation to waste generation is around 30% for Germany and 6% for Brazil. The FL recycling technologies lead up to fractions with different degrees of purity which impacts in the disposal of the materials. In Brazil, the main destination of the glass is for the ceramics industry, and for Germany it is used in the manufacture of glass tubes for new FL. Another difference is that Germany has already started the rare earth recovery from the fluorescent powder. The destination of mercury has several possibilities, due to costs of purification. The proposed model in this study for the reverse logistics of FL in Brazil considers the creation of a federal regulation setting specific targets for collection, recycling, criteria for system management, transportation, environmental labeling, limits of the use of mercury in lamps and control of the material destinations. The system framework consists of an association to manage the activities of collection, transportation and recycling, in order to reduce costs and complexity of the system as well as the foundation of an entity to record the amounts which are produced, imported, collected and recycled. Thus, through the implementation of reverse logistics based on the factors presented, it would become possible to increase the recycling rates, reducing environmental impacts and promoting the generation of new business.

In previous experiments, BACILA, ROSA, and RODRIGUES showed that enolase, partially purified from the flesh muscle of the krill Euphausia superba DANA, has not been inhibited by F^- in concentrations as high as 2 mM, a property which is paralleled by the enolase from the penguins liver. This finding led to the study of the structure and the kinetic properties of other enzymes from the carbohydrate metabolism of the krill's flesh muscle. In the present communication some data on the kinetic properties of the pyruvate kinase (PK) purified from the krill's flesh muscle are shown. L-Phenylalanine inhibits this preparation of PK both in the presence and in the absence of 1 mM L-alanine. However, the inhibition of PK by Mg-ATP was almost completely reversed in the presence of 1 mM Fru-P_2, but not in the presence of 1 mM L-alanine. The Km values for PK have been established to be 0.03 μM PEP and 0.038 μM PEP respectively in the presence and in the absence of Fru-P_2. This flesh muscle krill's preparation of PK shows clear sigmoidal kinetics. However, in the presence of Fru-P_2, PK shows a Michaelian kinetics. The effect of ADP has also been established and found to have a Km value of 0.15 μM ADP. Cations inhibit the krill's pyruvate kinase in the following order: Mn^> Ca^>Ni^>Co^ while Li^ did not show any inhibitory effect.

Phosphofructokinase (PFK) and hexokinase (HK) were concentrated by ammonium sulfate precipitation from the Trematomus bernacchii's epaxial and cardiac muscles respectively. The apparent Km of PFK for ATP and F6P was established at pH 8.0, at four different temperatures: 5°, 11°, 20° and 30℃. No significant effect of the temperature on the enzyme activity was observed, due to the fact that PFK does not exhibit allosteric properties at pH 8.0. However, at pH 7.0, in which condition the allosteric properties of PFK are displayed, it was observed that the rate of inhibition of the enzyme by ATP at 5℃ is lower than at 20℃. Kinetic data of the allosteric behavior of PFK at pH 7.0, were assayed at 5° and 20℃ in the presence of fixed concentrations of ATP and variable concentrations of F6P. According to the results obtained it was possible to conclude that the enzyme affinity with the substrate was 20 times higher at 5℃ than at 20℃. This result seems to possess high significance in regard to the mechanisms of biochemical adaptation of those organisms at low temperatures. Values for energy of activation for both PFK and HK were also established. The apparent KM for HK from the cardiac muscle was equal to 384μM and 599μM at 5℃ and 20℃, respectively.

1. The sublethal effects of Folidol 600, an organophosphate containing methyl parathion as its active principle, were analysed in the fish Callichthys callichthys. 2. After an intracoelomatic application of 500 mg/kg of Folidol 600, the behavior and the levels of plasmatic cholinesterase were analysed along the time, in acute experiments. 3. Plasma cholinesterase activity was inhibited by 90% after 4 hr from the initial time, keeping this level of inhibition for 4 days. 4. It recovered slowly in the next 8 to 10 days, reaching about 80–90% of the normal levels around the 35th day after the beginning of the experiment. 5. Many behavioral changes were observed in the first 96 hr, such as: lowering of motility with loss of equilibrium and frequent unnatural body positions; uncoordinated movements leading to incapacity of food intake; and increase in the levels of respiratory frequency.

1. 1. A survey has been carried out on the blood constituents of penguins from the Pygoscellidae family, Pygoscellis antartica , P. papua and P. adeliae , and of skuas ( Chataracta maccormicki ). 2. 2. Glucose, non-protein nitrogen compounds, proteins, lipids and inorganic compounds and the electrophoretic patterns for hemoglobin, serum proteins and lipoproteins were studied. 3. 3. The values obtained for glucose partition in the blood, glycosylated hemoglobin and non-protein nitrogen compounds, are discussed.

O estudo de cinética de liberação de metais pesados é uma importante ferramenta de diagnóstico ambiental de áreas contaminadas, pois determina, além dos teores acumulados liberados após tempos crescentes de equilíbrio, a taxa (velocidade) de dessorção desses poluentes para a solução do solo. Com o objetivo de avaliar a cinética de liberação de Pb de solos da área de mineração e metalurgia de metais pesados, no município de Adrianópolis (PR), vale do rio Ribeira, selecionaram-se oito solos, submetidos a diferentes formas de contaminação (solos 2, 4 e 5 - incorporação de resíduos da metalurgia de metais pesados aos perfis; solos 3, 6 e 7 - adição de Pb particulado via chaminés da fábrica; solo 8 - contaminação por passagem da água pluvial pela fábrica desativada, que escorre em direção ao rio Ribeira). O solo 1, sob mata nativa, fora da direção de caminhamento das fumaças da metalurgia, localizado a 1.560 m de distância e a 380 m acima da cota da fábrica desativada, foi escolhido como referência dos teores naturais de Pb dos solos da região. As amostras de solo, coletadas em duas profundidades (0 a 10 e 20 a 40 cm), foram submetidas a extrações sequenciais com ácido cítrico 0,1 mol L-1 após diferentes períodos de contato (tempos acumulados: 2, 14, 38, 86, 182, 326, 518, 806 e 1.382 h). Os teores totais de Pb (extração com HF e HNO3 concentrados e H2O2 30 % v/v) foram altos (máximo de 24.755,6 mg kg-1) e indicaram intensa contaminação dos solos. Os dados da cinética de liberação foram ajustados às equações parabólicas de difusão, sendo a liberação do Pb gradual e bifásica, com velocidade de liberação maior na primeira fase. Os solos 3 e 5 foram considerados com elevado potencial de contaminação ambiental devido aos altos teores de Pb dessorvido acumulados nas extrações sequenciais com ácido cítrico (máximo de 18.577,4 mg kg-1). O solo 6, com baixos teores de argila, apresentou a maior velocidade (taxa) de dessorção de Pb. Kinetics studies of heavy metal release are an important form of environmental diagnostics in contaminated areas, due to the possibility of determining, aside from the total release contents after increasing equilibrium times, the rate (velocity) of heavy metal to soil solution. The objective of this study was to determined Pb kinetics release from soils of the heavy metal mining and metallurgy area in Adrianópolis (PR), Brazil, in the Ribeira river valley. Eight soils were exposed to different contamination forms: (soils 2, 4 and 5 - residue incorporated in the soil profile; soils 3, 6 and 7 - sprinkled with particulate matter from the factory chimney; soil 8 - contamination by rain water flowing from the shut down factory towards the Ribeira river). Soil 1, under native forest, was chosen as reference of the natural soil Pb levels. The soil samples, collected from two layers (0-10 cm, 20-40 cm), were subjected to sequential extractions with 0.1 mol L-1 citric acid during different periods (2; 14; 38; 86; 182; 326; 518; 806; and 1,382 h). The high total Pb contents (extracted by concentrated HF and HNO3 and 30 % (v/v) H2O2) (maximum content of 24,755.6 mg kg-1) revealed intense soil contamination. The release kinetics data were fitted to parabolic equations of diffusion; lead release was gradual and biphasic (higher in the first phase). The environmental risks were considered high in the soils 3 and 5, where the evaluated levels of total Pb accumulated were high (reaching 18,577.4 mg kg-1). The Pb release rate was highest from soil 6, where clay content was low.

The purpose of this study was to compare the apical sealing capacity of three types of gutta-percha master cones of the same apical size and different tapers following root canal preparation with nickel–titanium ProTaper Universal rotary instruments and microstructural replication with System B and Obtura III. Thirty extracted human incisors having one single straight root canal (type I Weine) were instrumented with rotary ProTaper to an F3 (30/.09) and gauged to confirm a final apical size of #30. Teeth were divided into three groups (n = 10) to be obturated as follows: Group 1: master cone Meta 0.06 taper/AH Plus, Group 2: master cone fine-medium Autofit 0.08 taper/AH Plus, and Group 3: master cone ProTaper F3 0.09 taper/AH Plus. The chosen technique was the continuous wave of compaction (System B and Obtura III). Teeth were embedded in acrylic and incrementally reduced at 0.5 and 1.0 mm from the apical foramina in a grinding machine for metallographic samples. Sections were examined and digitally photographed under a metallographic optical microscope in normal and polarized light and the images were processed. The total cross-sectional area of the root canal, the gutta-percha/sealer/voids’ areas were quantified for each sample and statistically compared using one-way ANOVA and Kruskal–Wallis tests. No statistically significant differences between groups were observed (P > 0.05); however, the mean percentage of the gutta-percha-filled area was slightly higher in Group 1 at both levels of observation. Despite different tapers, all the three types of cones provided a good sealing capacity in the last apical millimeter of the root canal, with good gutta-percha–sealer ratio and few or no voids.

Secretaria de Estado de Ciencia Tecnologia e Ensino Superior (SETI) do Parana Fundacao Araucaria-PR Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Brown spider dermonecrotic toxins (phospholipases-D) are the most well-characterized biochemical constituents of Loxosceles spp. venom. Recombinant forms are capable of reproducing most cutaneous and systemic manifestations such as dermonecrotic lesions, hematological disorders, and renal failure. There is currently no direct confirmation for a relationship between dermonecrosis and inflammation induced by dermonecrotic toxins and their enzymatic activity. We modified a toxin isoform by site-directed mutagenesis to determine if phospholipase-D activity is directly related to these biological effects. the mutated toxin contains an alanine substitution for a histidine residue at position 12 (in the conserved catalytic domain of Loxosceles intermedia Recombinant Dermonecrotic Toxin - LiRecDT1). LiRecDT1H12A sphingomyelinase activity was drastically reduced, despite the fact that circular dichroism analysis demonstrated similar spectra for both toxin isoforms, confirming that the mutation did not change general secondary structures of the molecule or its stability. Antisera against whole venom and LiRecDT1 showed cross-reactivity to both recombinant toxins by ELISA and immunoblotting. Dermonecrosis was abolished by the mutation, and rabbit skin revealed a decreased inflammatory response to LiRecDT1H12A compared to LiRecDT1. Residual phospholipase activity was observed with increasing concentrations of LiRecDT1H12A by dermonecrosis and fluorometric measurement in vitro. Lipid arrays showed that the mutated toxin has an affinity for the same lipids LiRecDT1, and both toxins were detected on RAEC cell surfaces. Data from in vitro choline release and HPTLC analyses of LiRecDT1-treated purified phospholipids and RAEC membrane detergent-extracts corroborate with the morphological changes. These data suggest a phospholipase-D dependent mechanism of toxicity, which has no substrate specificity and thus utilizes a broad range of bioactive lipids. (C) 2010 Elsevier B.V. All rights reserved. Univ Fed Parana, Dept Cell Biol, BR-81531990 Curitiba, Parana, Brazil Universidade Federal de São Paulo, Dept Biochem, São Paulo, Brazil Univ Estadual Ponta Grossa, Dept Struct Mol Biol & Genet, Ponta Grossa, Brazil Catholic Univ Parana, Hlth & Biol Sci Inst, Curitiba, Parana, Brazil Universidade Federal de São Paulo, Dept Biochem, São Paulo, Brazil Web of Science

The aim of this work was to produce and characterize monoclonal antibodies anti bovine immunoglobulin G (IgG). Out of seven hybridomas, two were chosen based on the ELISA'S absorbance values and were labeled B4F11 and B3H12. These monoclonals were analyzed through Western Blot for IgG fragments obtained by proteolysis with papain, separated by electrophoresis in polyacrylamide gel electrophoresis with β-mercaptoetanol as reducing agent. This revealed that, possibly, the B4F11 was directed to a conformational antigen, and that B3H12 reacted in a specific fashion with Fc (Bovine IgG crystallizable fragment). This antibody could be used in the development of reagents to immunoassays relevant for research and diagnosis.No Brasil, anticorpos anti-espécie específica usados em métodos de diagnóstico geralmente são importados, aumentando o custo das análises. Visando produzir insumos para métodos diagnóstico por enzimaimunoensaio, o presente trabalho teve como objetivo produzir e caracterizar anticorpos monoclonais anti imunoglobulina G (IgG) bovina. De sete hibridomas obtidos e que apresentaram valores relevantes de absorbância em teste imunoenzimático (ELISA indireta), dois clones com melhor performance foram selecionados e designados B4F11 e B3H12. Estes anticorpos monoclonais foram analisados em Western Blot para reatividade com fragmentos de IgG bovina, obtidos por proteólise com papaína e separados por eletroforese em gel de poliacrilamida, com presença do agente redutor beta-mercaptoetanol. A eletroforese mostrou que o anticorpo B4F11, foi direcionado para um antígeno conformacional, e que o monoclonal B3H12 reagiu especificamente com a porção Fc da IgG bovina (fragmento cristalizável). Este anticorpo será utilizado no desenvolvimento de reagentes para imunoensaios de interesse à pesquisa e diagnósticos.

1. 1. Pyruvate kinase from the striated muscle of Chaenocephalus aceratus was purified to homogeneity by a 3-step process, 1 step involving ammonium sulfate precipitation and 2 steps with phosphocellulose and KC1 and phosphoenolpyruvate elution. The final specific activity of the enzyme was 188.8 UI/mg and the overall yield 20%. The apparent molecular weights of the native enzyme suggests that the protein is present as a homotetramer. 2. 2. The cellulose acetate electrophoresis showed the presence of only 1 isoenzymatic form with electrophoretic mobility similar to that of the L type PK from rat liver. 3. 3. The enzyme possesses an optimal pH around 6.5 and an energy of activation of 15,000 cal/mol. Among 6 different buffers used, imidazole-HCl was the only one which did not interfere with the enzyme activity. Furthermore, this buffer showed to be the most efficient one in stabilizing the PK at 40°C for 30 min.