Polymerase chain reaction tests for the identification of Ross River, Kunjin and Murray Valley encephalitis virus infections in horses

Objective To develop and validate specific, sensitive and rapid diagnostic tests using RT-PCR for the detection of Ross River virus (RRV), Kunjin virus (KV) and Murray Valley encephalitis virus (MVEV) infections in horses. Methods Primer sets based on nucleotide sequence encoding the envelope glycoprotein E2 of RRV and on the nonstructural protein 5 (NS5) of KV and MVEV were designed and used in single round PCRs to test for the respective viruses in infected cell cultures and, in the case of RRV, in samples of horse blood and synovial fluid. Results The primer pairs designed for each of the three viruses amplified a product of expected size from prototype viruses that were grown in cell culture. The identity of each of the products was confirmed by nucleotide sequencing indicating that in the context used the RT-PCRs were specific. RRV was detected in serums from 8 horses for which there were clinical signs consistent with RRV infection such that an acute-phase serum sample was taken and submitted for RRV serology testing. The RRV RT-PCR was analytically sensitive in that it was estimated to detect as little as 50 TCID50 of RRV per mL of serum and was specific in that the primer pairs did not amplify other products from the 8 serum samples. The RRV primers also detected virus in three independent mosquito pools known to contain RRV by virus isolation in cell culture. Samples from horses suspected to be infected with KV and MVEV were not available. Conclusion Despite much anecdotal and serological evidence for infection of horses with RRV actual infection and associated clinical disease are infrequently confirmed. The availability of a specific and analytically sensitive RT-PCR for the detection of RRV provides additional opportunities to confirm the presence of this virus in clinical samples. The RTPCR primers for the diagnosis of KV and MVEV infections were shown to be specific for cell culture grown viruses but the further validation of these tests requires the availability of appropriate clinical samples from infected horses.

Variation of bulk properties of anaerobic granules with wastewater type

Development of a granular sludge with high strength, high biological activity and a narrow settling distribution is necessary for optimal operation of high-rate upflow anaerobic treatment systems. Several studies have compared granules produced from different wastewaters but these have largely been from laboratory-fed reactors or compared granules from full-scale reactors fed similar wastewater types. Though two authors have commented on the inferiority of granules produced by a protein-based feed, the properties of these granules have not been characterised. In this paper, granules from full-scale reactors treating fruit and vegetable cannery effluent, two brewery effluents and a pig abattoir (slaughterhouse) were compared in terms of basic composition, size distribution, density, settling velocity, shear strength, and EPS content. The results supported previous qualitative observations by other researchers that indicate granule properties depend more on wastewater type rather than reactor design or operating conditions such as pre-acidification level. The cannery-fed granules bad excellent shear strength, settling distribution and density. Granules from the two brewery-fed reactors had statistically the same bulk properties, which were still acceptable for upflow applications. The protein-grown granule had poor strength and settling velocity. (C) 2001 Elsevier Science Ltd. All rights reserved.

Invertebrate D2 type dopamine receptor exhibits age-based plasticity of expression in the mushroom bodies of the honeybee brain

We have isolated a cDNA clone from the honeybee brain encoding a dopamine receptor, AmDop2, which is positively coupled to adenylyl cyclase. The transmembrane domains of this receptor are 88% identical to the orthologous Drosophila D2 dopamine receptor, DmDop2, though phylogenetic analysis and sequence homology both indicate that invertebrate and vertebrate D2 receptors are quite distinct. In situ hybridization to mRNA in whole-mount preparations of honeybee brains reveals gene expression in the mushroom bodies, a primary site of associative learning. Furthermore, two anatomically distinct cell types in the mushroom bodies exhibit differential regulation of AmDop2 expression. In all nonreproductive females (worker caste) and reproductive males (drones) the receptor gene is strongly and constitutively expressed in all mushroom body interneurons with small cell bodies. In contrast, the large cell-bodied interneurons exhibit dramatic plasticity of AmDop2 gene expression. In newly emerged worker bees (cell-cleaning specialists) and newly emerged drones, no AmDop2 transcript is observed in the large interneurons whereas this transcript is abundant in these cells in the oldest worker bees (resource foragers) and older drones. Differentiation of the mushroom body interneurons into two distinct classes (i.e., plastic or nonplastic with respect to AmDop2 gene expression) indicates that this receptor contributes to the differential regulation of distinct neural circuits. Moreover, the plasticity of expression observed in the large cells implicates this receptor in the behavioral maturation of the bee.

Early pregnancy factor treatment suppresses the inflammatory response and adhesion molecule expression in the spinal cord of SJL/J mice with experimental autoimmune encephalomyelitis and the delayed-type hypersensitivity reaction to trinitrochlorobenzene in normal BALB/c mice

Early pregnancy factor (EPF) is a secreted protein, present in serum during early pregnancy and essential for maintaining viability of the embryo. It is a homologue of chaperonin 10 (Cpn10) but, unlike Cpn10, it has an extracellular role. EPF has immunosuppressive and growth regulatory properties. Previously we have reported the preparation of recombinant EPF (rEPF) and shown that treatment with rEPF will suppress clinical signs of MBP-EAE in Lewis rats and PLP-EAE in SJL/J mice. In the present study, these findings have been extended to investigate possible mechanisms involved in the action of EPF. Following treatment of mice with rEPF from the day of inoculation, there were fewer infiltrating CD3+ and CD4+ cells in the parenchyma of the spinal cord during the onset of disease and after the initial episode, compared with mice treated with vehicle. Expression of the integrins LFA-1, VLA-4 and Mac-1 and of members of the immunoglobulin superfamily of adhesion molecules ICAM-1 and VCAM-1 was suppressed in the central nervous system (CNS) following rEPF treatment. The expression of PECAM-1 was not affected. To determine if rEPF suppressed T cell activation in the periphery, the delayed-type hypersensitivity (DTH) reaction of normal BALB/c mice to trinitrochlorobenzene (TNCB) following treatment with rEPF was studied. The results showed that treatment with rEPF suppressed the DTH reaction, demonstrating the ability of EPF to downregulate the cell-mediated immune response. These results indicate that suppression of immunological mechanisms by rEPF plays a major role in the reduction of clinical signs of disease in experimental autoimmune encephalomyelitis (EAE). (C) 2003 Elsevier Science B.V. All rights reserved.

Usefulness of B-type natriuretic peptide in hypertensive patients with exertional dyspnea and normal left ventricular ejection fraction and correlation with new echocardiographic indexes of systolic and diastolic function

B-type natriuretic peptide (BNP) levels increase in systolic heart failure (HF). However, the value of BNP in hypertensive patients with suspected diastolic HF (symptoms suggestive of HF but normal ejection fraction) and its relation to myocardial function in these patients is unclear. We prospectively studied 72 ambulatory hypertensive subjects (40 women, mean age 58 +/- 8 years) with exertional dyspnea and ejection fraction greater than or equal to50%. Diastolic function was evaluated with transmitral and pulmonary venous Doppler, mitral annular velocities (pulsed-wave tissue Doppler), and flow propagation velocity (color M-mode). Systolic function was assessed with strain and strain rate derived from color tissue Doppler imaging. BNP was related to myocardial function and the presence or absence of global diastolic dysfunction. By conventional Doppler criteria, 34 patients had normal left ventricular diastolic function and 38 had isolated diastolic dysfunction. BNP values were higher in patients with diastolic dysfunction (46 +/- 48 vs 20 +/- 20 pg/ml, p = 0.004) and were related independently to blood pressure, systolic strain rate, left atrial function (p < 0.01 for all), and age (p = 0.015). Patients with diastolic dysfunction and pseudonormal filling had higher BNP levels compared with impaired relaxation (89 +/- 47 vs 35 +/- 42 pg/ml, p = 0.001). However, 79% of patients with diastolic dysfunction had BNP levels within the normal range. We conclude that in ambulatory hypertensive patients with symptoms suggestive of mild HF and normal ejection fraction, BNP is related to atrial and ventricular systolic parameters, blood pressure, and age. Although elevated in the presence of diastolic dysfunction, the BNP level mostly is in the normal range and, therefore, has limited diagnostic value in stable patients with suspected diastolic HF. (C) 2003 by Excerpta Medica, Inc.

Structure of the HERG K+ channel S5P extracellular linker - Role of an amphipathic alpha-helix in C-type inactivation

The HERG K+ channel has very unusual kinetic behavior that includes slow activation but rapid inactivation. These features are critical for normal cardiac repolarization as well as in preventing lethal ventricular arrhythmias. Mutagenesis studies have shown that the extracellular peptide linker joining the fifth transmembrane domain to the pore helix is critical for rapid inactivation of the HERG K+ channel. This peptide linker is also considerably longer in HERG K+ channels, 40 amino acids, than in most other voltage-gated K+ channels. In this study we show that a synthetic 42-residue peptide corresponding to this linker region of the HERG K+ channel does not have defined structural elements in aqueous solution; however, it displays two well defined helical regions when in the presence of SDS micelles. The helices correspond to Trp(585)-Ile(593) and Gly(604)-Tyr(611) of the channel. The Trp(585)-Ile(593) helix has distinct hydrophilic and hydrophobic surfaces. The Gly(604)-Tyr(611) helix corresponds to an N-terminal extension of the pore helix. Electrophysiological studies of HERG currents following application of exogenous S5P peptides show that the amphipathic helix in the S5P linker interacts with the pore region of the channel in a voltage-dependent manner.

Effect of central venous catheter type on infections: a prospective clinical trial

This study reports on a block clinical trial of two types of central venous catheters (CVCs): antiseptic-impregnated catheters (AIC) and non-impregnated catheters (non-AIC), on catheter tip colonization and bacteraemia. In total, 500 catheters were inserted in 390 patients over the 18 month study period, 260 (52.0%) AIC and 240 (48.0%) non-AIC. Of these, 460 (92.0%) tips (237 AIC and 223 non-AIC) were collected. While significantly fewer AIC, 14 (5.9%), than non-AIC, 30 (13.5%), catheters were colonized (P < 0.01), there was no difference in the rates of bacteraemias in the two groups (0.8% vs. 2.7%, respectively, P = 0.16). There were 6.87 (95% CI 3.38-14.26) and 16.92 (95% CI 10.61-27.12) colonized AIC and non-AIC catheters, respectively, per 1000 catheter days, a difference that was significant (P < 0.01). However, no difference emerged between bacteraemias in AIC and non-AIC catheters per 1000 catheter days measured at 0.98 (95% CI 0.24-5.54) and 3.38 (95% CI 1.29-9.34), respectively (P = 0.10). Of the 444 CVCs that were sited in the subclavian or jugular veins and had tips collected, significantly more catheters were colonized in the jugular group, 19 (20%), compared with the subclavian group, 24 (6.9%; P less than or equal to 0.01). Overall, the low rates of colonization and bacteraemia may be explained by the population studied, the policies used and the employment of a clinical nurse dedicated to CVC management. (C) 2003 The Hospital Infection Society. Published by Elsevier Science Ltd. All rights reserved.

Honorários, serviços fora da área de auditoria e independência do auditor : evidências nas companhias abertas listadas na BM&FBovespa

O presente trabalho possui o objetivo de identificar os determinantes dos honorários e serviços fora da área de auditoria, bem como avaliar se a prestação destes serviços conjuntamente com a auditoria das demonstrações financeiras afeta a independência dos auditores externos das companhias abertas brasileiras negociadas na BM&FBovespa. A amostra contempla um total de 180 empresas não financeiras observadas no período de 2010 a 2012. Os resultados indicam que as proxies para esforço e risco elevam os honorários de auditoria, enquanto a governança corporativa apresentou tanto o efeito demanda (elevam os honorários) quanto o efeito risco (reduzem os honorários) dependendo da proxy observada. No que tange a contratação de serviços fora da área de auditoria foi constatado que empresas com maior necessidade de monitoramento e com comitês de auditoria demandam esses serviços, embora o conselho fiscal iniba a contratação dos mesmos. Por fim, as evidências indicam que a prestação de serviços fora da área de auditoria não afeta a independência do auditor. Estes resultados são mantidos mesmo quando observada a probabilidade de ocorrência da modificação no parecer da auditoria relacionada à violação da premissa de continuidade da firma (Going Concern Opinion). Os testes adicionais realizados confirmam a manutenção da independência do auditor, inclusive quando observado o tipo de auditor (não Big4), expectativa de ganhos futuros relacionados a serviços fora da área de auditoria adicionais, empresas consideradas ressalváveis, bem como o efeito de melhores práticas de governança corporativa.

The need for standardized tests to evaluate the reliability of data transport in wireless medical systems

Wireless medical systems are comprised of four stages, namely the medical device, the data transport, the data collection and the data evaluation stages. Whereas the performance of the first stage is highly regulated, the others are not. This paper concentrates on the data transport stage and argues that it is necessary to establish standardized tests to be used by medical device manufacturers to provide comparable results concerning the communication performance of the wireless networks used to transport medical data. Besides, it suggests test parameters and procedures to be used to produce comparable communication performance results.

Carbon Nanofiber Type and Content Dependence of the Physical Properties of Carbon Nanofiber Reinforced Polypropylene Composites

The variation of the physical properties of four differ- ent carbon nanofibers (CNFs), based-polymer nano- composites incorporated in the same polypropylene (PP) matrix by twin-screw extrusion process was investigated. Nanocomposites fabricated with CNFs with highly graphitic outer layer revealed electrical isolation-to-conducting behaviors as function of CNF’s content. Nanocomposites fabricated with CNFs with an outer layer consisting on a disordered pyro- litically stripped layer, in contrast, revealed better mechanical performance and enhanced thermal sta- bility. Further, CNF’s incorporation into the polymer increased the thermal stability and the degree of crystallinity of the polymer, independently on the filler content and type. In addition, dispersion of the CNFs’ clusters in PP was analyzed by transmitted light opti- cal microscopy, and grayscale analysis (GSA). The results showed a correlation between the filler concentration and the variance, a parameter which measures quantitatively the dispersion, for all composites. This method indicated a value of 1.4 vol% above which large clusters of CNFs cannot be dispersed effectively and as a consequence only slight changes in mechanical performance are observed. Finally, this study establishes that for tailoring the physical properties of CNF based-polymer nanocomposites, both adequate CNFs structure and content have to be chosen.

Type checking cryptography implementations

Cryptographic software development is a challenging eld: high performance must be achieved, while ensuring correctness and com- pliance with low-level security policies. CAO is a domain speci c language designed to assist development of cryptographic software. An important feature of this language is the design of a novel type system introducing native types such as prede ned sized vectors, matrices and bit strings, residue classes modulo an integer, nite elds and nite eld extensions, allowing for extensive static validation of source code. We present the formalisation, validation and implementation of this type system

Diagnostic performance of visual screening tests in the elderly

This study aimed to determine and evaluate the diagnostic accuracy of visual screening tests for detecting vision loss in elderly. This study is defined as study of diagnostic performance. The diagnostic accuracy of 5 visual tests -near convergence point, near accommodation point, stereopsis, contrast sensibility and amsler grid—was evaluated by means of the ROC method (receiver operating characteristics curves), sensitivity, specificity, positive and negative likelihood ratios (LR+/LR−). Visual acuity was used as the reference standard. A sample of 44 elderly aged 76.7 years (±9.32), who were institutionalized, was collected. The curves of contrast sensitivity and stereopsis are the most accurate (area under the curves were 0.814−p = 0.001, C.I.95%[0.653;0.975]— and 0.713−p = 0.027, C.I.95%[0,540;0,887], respectively). The scores with the best diagnostic validity for the stereopsis test were 0.605 (sensitivity 0.87, specificity 0.54; LR+ 1.89, LR−0.24) and 0.610 (sensitivity 0.81, specificity 0.54; LR+1.75, LR−0.36). The scores with higher diagnostic validity for the contrast sensibility test were 0.530 (sensitivity 0.94, specificity 0.69; LR+ 3.04, LR−0.09). The contrast sensitivity and stereopsis test's proved to be clinically useful in detecting vision loss in the elderly.

Diagnostic performance of visual screening tests in the elderly

The aging of Portuguese population is characterized by an increase of individuals aged older than 65 years. Preventable visual loss in older persons is an important public health problem. Tests used for vision screening should have a high degree of diagnostic validity confirmed by means of clinical trials. The primary aim of a screening program is the early detection of visual diseases. Between 20% and 50% of older people in the UK have undetected reduced vision and in most cases is correctable. Elderly patients do not receive a systematic eye examination unless a problem arises with their glasses or suspicion vision loss. This study aimed to determine and evaluate the diagnostic accuracy of visual screening tests for detecting vision loss in elderly. Furthermore, it pretends to define the ability to find the subjects affected with vision loss as positive and the subjects not affected with the same disease as negative. The ideal vision screening method should have high sensitivity and specificity for early detection of risk factors. It should be also low cost and easy to implement in all geographic and socioeconomic regions. Sensitivity is the ability of an examination to identify the presence of a given disease and specificity is the ability of the examination to identify the absence of a given disease. It was not an aim of this study to detect abnormalities that affect visual acuity. The aim of this study was to find out what´s the best test for the identification of any vision loss.

Ash deposition during the co-firing of bituminous coal with pine sawdust and olive stones in a laboratory furnace

This article describes an experimental study on ash deposition during the co-firing of bituminous coal with pine sawdust and olive stones in a laboratory furnace. The main objective of this study was to relate the ash deposit rates with the type of biomass burned and its thermal percentage in the blend. The thermal percentage of biomass in the blend was varied between 10% and 50% for both sawdust and olive stones. For comparison purposes, tests have also been performed using only coal or only biomass. During the tests, deposits were collected with the aid of an air-cooled deposition probe placed far from the flame region, where the mean gas temperature was around 640 degrees C. A number of deposit samples were subsequently analyzed on a scanning electron microscope equipped with an energy dispersive X-ray detector. Results indicate that blending sawdust with coal decreases the deposition rate as compared with the firing of unblended coal due to both the sawdust low ash content and its low alkalis content. The co-firing of coal and sawdust yields deposits with high levels of silicon and aluminium which indicates the presence of ashes with high fusion temperature and, thus, with less capacity to adhere to the surfaces. In contrast, in the co-firing of coal with olive stones the deposition rate increases as compared with the firing of unblended coal and the deposits produced present high levels of potassium, which tend to increase their stickiness.