Deliriumin the ICU setting ‐ a subjective and theoretical survey before the implementation of the Confusion Assessment Method for the ICU in an unit

Background and Goal of Study: The current definition of delirium comprises acute change or fluctuation in mental status and inattention, accompanied by either altered level of consciousness or disorganized thinking. It is a frequent condition in the ICU and it is associated with longer hospital stay, increase in mortality at 6 months and long-term cognitive impairment, but remains under diagnosed. The Confusion Assessment Method for the Intensive Care Unit (CAM-ICU) has been validated and implemented in many ICUs and its use is recommended by the Society of Critical Care Medicine. It is our purpose to evaluate the individual perspective and the objective knowledge of our staf f about delirium before the implementation of the CAM-ICU. Materials and Methods: Anonymous survey to our ICU clinical staf f which contained subjective and ‘true or false’ questions. Data was analised with the sof tware SPSS version 17.0. The Wilcoxon test was used to compare autoperception of knowledge about delirium and the content of answers regarding its definiton. Results: Forty two questionnaires were returned (participation rate of 73%), 11 from physicians and 31 from nurses. Overall, 61,9% of inquiries think they can give a definition for delirium in the ICU and 50% claim to be able to evaluate delirium. 28,6% of the respondents - 63,6% of the physicians and 16,1% of the nurses - know the CAM-ICU. From these only a quarter has received education on this method, 75% think it’s easy to apply and 66% don’t see its use as an increase in the daily workload. We found a high rate of wrong and ‘I don’t know’ answers to questions about operationalization, diagnosis and outcome. The subjects’ auto-perception on their knowledge about delirium [Likert scale] was compared to their ability to answer questions related to its definiton - ‘attention deficit is essencial for diagnosis’ [true], Wilcoxon test Z=-4,699 (p< 0,001); ‘disorganized thinking is essential for diagnosis’ [false], Wilcoxon test Z=-4,437 (p< 0,001). Conclusions: The respondents’ auto perception of knowledge about delirium doesn’t translate in the ability of giving an appropriate definition and making an adequate evaluation. Most of the inquiries don’t know the CAM-ICU, but those who do believe it’s easy to apply and its use won’t increase the workload. We performed educational sessions about delirium and the CAM-ICU in our unit to encourage our clinical staf f to deal properly with this hazardous condition

Sustained-Release Bupropion Overdose: A Case Report

Bupropion is an atypical antidepressant with a unique aminoketone structure similar to amphetamines. A narrow therapeutic margin is evident from observational studies that show seizure activity with doses of 400-600 mg or higher. A 38-year old woman took an overdose of 6 grams of bupropion with 110 grams of alcohol. She presented to the Emergency Department with agitation, visual hallucinations and myoclonus of the upper limbs; eyes spontaneously open with isochoric and light reactive pupils with horizontal nystagmus; afebrile, normotensive (121/63 mm Hg) and tachycardic (120 beats/minute). The electrocardiogram revealed a sinus tachycardia with prolonged QT interval (QT/QTc: 0.46/ 0.537) and a QRS complex length in the upper limit of normal. Arterial blood gases revealed metabolic acidosis (pH = 7.16) with increased anion-gap (value=18). She developed mal epilepticus needing thiopental induced coma and Intensive Care Unit (ICU) admission. She suffered prolonged symptoms including seizures before fully recovering. The narrow therapeutic range and the increasing use in the treatment of smoking cessation boosted the number of intentional and unintentional poisoning by this drug. Previous reports of bupropion overdose almost all involve the immediate release formulation. There are some reports of overdose with sustained-release formulation, but there is limited information on its spectrum of toxicity

Método do Dot-Elisa para diagnóstico da Maedi-Visna.

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Maedi-visna vírus: produção de antígeno, análise protéica e antigênica.

A Maedi-Visna é uma doença persistente, progressiva e debilitante em ovinos causada por lentivírus que resulta primariamente em pneumonia intersticial e pode estar associada a mamite e encefalite. Este trabalho teve como objetivo produzir antígeno a partir do vírus Maedi-Visna total para utilização em ensaios imunoenzimáticos. Na produção do antígeno foram utilizados cultivos primários de células de membrana sinovial caprina, infectados com amostra padrão (MVVK1514). As suspensões virais foram tituladas e o antígeno semipurificado pela precipitação em PEG (polietilenoglicol) e ultracentrifugação. Foram realizadas eletroforese em gel de poliacrilamida (SDS-PAGE) e Western Blotting (WB). A SDS-PAGE de origem viral e do meio de cultivo resultou em várias bandas protéicas. No WB constatou-se a presença de sete proteínas imunogênicas de pesos moleculares aproximados de 16, 27, 35, 50, 42, 63 e 123 kDa. Destas, três proteínas (16, 27 e 50 kDa) apresentaram boa reação imunogênica. O trabalho abre perspectivas da utilização de testes imunoenzimáticos com maior sensibilidade para lentivírus ovino. [Maedi-Visna virus: antigen production, protein and antigenic analysis]. Abstract - Maedi-Visna is a persistent, progressive and debilitating disease in sheep caused by lentivirus which results primarily in interstitial pneumonia and may be associated with mastitis and encephalitis. This study aimed to produce antigen from the Maedi-Visna virus for use in immunosorbent assay. The antigen was produced using secondary cultures of cells from sheep synovial membrane, infected with the standard sample (MVV-K1514). The suspensions were titered and the viral antigen was semipurified by precipitation in PEG (polyethylene) and ultracentrifugation. The isolates were then submitted to polyacrylamide gel electrophoresis (SDSPAGE) and the western blot (WB) technique. The SDS-PAGE of viral origin and of the culture medium resulted in several protein bands. The WB revealed the presence of 7 immunogenic proteins of approximate molecular weights of 16, 27, 35, 50, 42, 63 and 123 kDa. Of these, 3 proteins (16, 27 and 50 kDa) showed good immunogenic reaction. This work opens up prospects of using enzyme immunoassay with greater sensitivity to lentivirus-infected sheep

Isolation and identification of caprine lentivirus in synovial fluid and blood using Nested PCR and western blotting.

The caprine arthritis-encephalitis (CAE) is an infectious, multisystem disease, caused by a retrovirus belongs to the Lentivirus genus, which affects all ages goats and breed types. This study aimed to investigatethe caprine arthritis-encephalitis virus (CAEV)through the isolation of native strains and identification with polymerase chain reaction (nested PCR) and western bloting (WB)

Frequência de anticorpos contra o vírus da língua azul em ovinos do Estado do Ceará, Brasil.

Resumo: O objetivo deste trabalho foi avaliar a ocorrência de ovinos soropositivos para o vírus da língua azul (VLA) no Estado do Ceará, Brasil, e analisar as proteínas imunogênicas das cepas virais circulantes nesses rebanhos. O teste de imunodifusão em gel de agarose (IDGA) foi utilizado para pesquisar 271 amostras de soro oriundas de 16 rebanhos. Os resultados demonstraram que 27,3% (74/271) das amostras analisadas apresentaram anticorpos contra o agente e 68,8% (11/16) das propriedades tiveram animais positivos. O immunoblotting (IB) foi utilizado para analisar as proteínas imunogênicas do VLA a partir dos soros de animais positivos no IDGA. Os soros demonstraram forte reação contra a proteína viral VP2. Para o VLA, das sete proteínas estruturais, a VP2 é a principal a estimular a resposta imune protetora. Concluiu-se que a soropositividade para a língua azul (LA) nos rebanhos ovinos estudados no Ceará é alta, apesar dos animais não apresentarem sinais clínicos, indicativo de que o vírus ocorra de forma endêmica. Além disso, a resistência à doença apresentada pelos animais pode estar relacionada com a forte reação imunológica desses à proteína VP2. Sendo assim, outros estudos são necessários para melhor esclarecer a situação epidemiológica da LA no país, através da identificação dos vetores e sorotipos virais circulantes nas diferentes regiões. [Antibodies against the bluetongue virus in sheep flocks of Ceará State, Brazil.]. Abstract: The objective of this work was to verify the occurrence of sheep serologically positive for bluetongue virus (BTV) in the state of Ceará, Brazil, and analyze immunogenic proteins of circulating viral strains in these flocks. The agar gel immunodifusion test (AGID) was used to examine 271 serum samples from 16 herds. The results demonstrated that 27.3% (74/271) of the analyzed samples presented antibodies for the agent, and that 68.8% (11/16) of the properties presented positive animals. Immunoblotting (IB) was used to analyze the immunogenic proteins of BTV derived from AGID positive sera. Sera showed strong reaction against viral protein VP2. Of the seven BTV structural proteins, VP2 is the major protein to elicit protective immune responses. It was concluded that bluetongue (BT) seropositivity in sheep flocks studied in Ceará is high, despite that the animal's do not show clinical signs, indicating that it occurs in an endemic form. The animals? resistance to the disease may be related to the strong immune response to the protein VP2. Therefore, further studies are needed to better clarify the epidemiological situation of BT in Brazilian sheep flocks, through the identification of viral vectors and serotypes circulating in different regions

Desenvolvimento e padronização de um ELISA indireto para diagnóstico de Maedi Visna em ovinos.

O objetivo deste trabalho foi desenvolver e padronizar um ELISA indireto para diagnóstico de Maedi Visna (MV). Produziu-se o antígeno em sobrenadantes de cultivo celular de membrana sinovial caprina (MSC) inoculado com o Maedi Visna Vírus (MVV) cepa K1514, que passou por ciclos de congelamento e descongelamento, sendo logo após clarificado por centrifugação a 3.000 g por 40 minutos A suspensão clarificada foi precipitada por PEG 8000, centrifugada a 12.000 g por 60 minutos, o pellet ressuspendido em TNE (Tampão Tris-HCl, NaCl, EDTA) e ultracentrifugado a 42.000 g por 105 minutos em colchão de sacarose, e ressuspendido em PBS contendo phenylmethylsulphonyl fluoride (PMSF). Realizou-se o ELISA em microplacas de 96 poços, incubadas por 1h a 37 ºC, utilizando-se como revelador o-phenylenediamine (OPD). Para a comparação entre os testes de ELISA e AGID, utilizaram-se 175 amostras de soros. A concentração ótima do antígeno foi a de 2 µg/mL e a melhor diluição dos soros, controles e testes de 1:100. O ELISA detectou um maior número de positivos (40) que o AGID (11), apresentando uma sensibilidade de 91%, especificidade de 82%. O ELISA promoveu uma melhor sensibilidade que o AGID. Embora sua especificidade tenha ficado abaixo do esperado, seu uso pode ser indicado como teste de diagnóstico de MVV

Avanços recentes em nutrição de larvas de peixes

Os requisitos nutricionais de larvas de peixes são ainda mal compreendidos, o que leva a altas mortalidades e problemas de qualidade no seu cultivo. Este trabalho pretende fazer uma revisão de novas metodologias de investigação, tais como estudos com marcadores, genómica populacional, programação nutricional, génomica e proteómica funcionais, e fornecer ainda alguns exemplos das utilizações presentes e perspectivas futuras em estudos de nutrição de larvas de peixes