Primary hemocyte culture of Penaeus monodon as an in vitro model for white spot syndrome virus titration, viral and immune related gene expression and cytotoxicity assays

Immortal cell lines have not yet been reported from Penaeus monodon, which delimits the prospects of investigating the associated viral pathogens especially white spot syndrome virus (WSSV). In this context, a method of developing primary hemocyte culture from this crustacean has been standardized by employing modified double strength Leibovitz-15 (L-15) growth medium supplemented with 2% glucose, MEM vitamins (1 ), tryptose phosphate broth (2.95 g l 1), 20% FBS, N-phenylthiourea (0.2 mM), 0.06 lgml 1 chloramphenicol, 100 lgml 1 streptomycin and 100 IU ml 1 penicillin and hemolymph drawn from shrimp grown under a bio-secured recirculating aquaculture system (RAS). In this medium the hemocytes remained viable up to 8 days. 5-Bromo-20-deoxyuridine (BrdU) labeling assay revealed its incorporation in 22 ± 7% of cells at 24 h. Susceptibility of the cells to WSSV was confirmed by immunofluoresence assay using a monoclonal antibody against 28 kDa envelope protein of WSSV. A convenient method for determining virus titer as MTT50/ml was standardized employing the primary hemocyte culture. Expression of viral genes and cellular immune genes were also investigated. The cell culture could be demonstrated for determining toxicity of a management chemical (benzalkonium chloride) by determining its IC50. The primary hemocyte culture could serve as a model for WSSV titration and viral and cellular immune related gene expression and also for investigations on cytotoxicity of aquaculture drugs and chemicals

A New Model For Reliability Centered Maintenance In Petroleum Refineries

Refiners today operate their equipment for prolonged periods without shutdown. This is primarily due to the increased pressures of the market resulting in extended shutdown-to-shutdown intervals. This places extreme demands on the reliability of the plant equipment. The traditional methods of reliability assurance, like Preventive Maintenance, Predictive Maintenance and Condition Based Maintenance become inadequate in the face of such demands. The alternate approaches to reliability improvement, being adopted the world over are implementation of RCFA programs and Reliability Centered Maintenance. However refiners and process plants find it difficult to adopt this standardized methodology of RCM mainly due to the complexity and the large amount of analysis that needs to be done, resulting in a long drawn out implementation, requiring the services of a number of skilled people. These results in either an implementation restricted to only few equipment or alternately, one that is non-standard. The paper presents the current models in use, the core requirements of a standard RCM model, the alternatives to classical RCM, limitations in the existing model, classical RCM and available alternatives to RCM and will then go on to present an ‗Accelerated‘ approach to RCM implementation, that, while ensuring close conformance to the standard, does not place a large burden on the implementers

Standardized test to evaluate numerical weather prediction algorithms

In order to assist in comparing the computational techniques used in different models, the authors propose a standardized set of one-dimensional numerical experiments that could be completed for each model. The results of these experiments, with a simplified form of the computational representation for advection, diffusion, pressure gradient term, Coriolis term, and filter used in the models, should be reported in the peer-reviewed literature. Specific recommendations are described in this paper.

A tool for standardized collector performance calculations including PVT

A tool for standardized calculation of solar collector performance has been developed in cooperation between SP Technical Research Institute of Sweden, DTU Denmark and SERC Dalarna University. The tool is designed to calculate the annual performance of solar collectors at representative locations in Europe. The collector parameters used as input in the tool are compiled from tests according to EN12975, without any intermediate conversions. The main target group for this tool is test institutes and certification bodies that are intended to use it for conversion of collector model parameters (derived from performance tests) into a more user friendly quantity: the annual energy output. The energy output presented in the tool is expressed as kWh per collector module. A simplified treatment of performance for PVT collectors is added based on the assumption that the thermal part of the PVT collector can be tested and modeled as a thermal collector, when the PV electric part is active with an MPP tracker in operation. The thermal collector parameters from this operation mode are used for the PVT calculations. © 2012 The Authors.

Open and Standardized Resources for Smart Transducer Networking

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Fracture strength of bovine incisors after intra-radicular treatment with MTA in an experimental immature tooth model

Aim To evaluate, using an experimental immature tooth model, the fracture resistance of bovine incisors submitted to different reinforcement treatments with mineral trioxide aggregate (MTA).Methodology An immature tooth model was created by sectioning the coronal and apical portions of 40 bovine incisors 8 mm above and 12 mm below the cementoenamel junction. The root canals were irrigated with 1.0% sodium hypochlorite. They were enlarged both coronally and apically using number 703 carbide burs (ISO: 500-104-168-007-021) and their internal diameter was standardized to 2.1 mm. The specimens were assigned to four groups (n = 10): GI-control (without filling); GII-apical MTA plug + filling with gutta-percha and endodontic sealer; GIII-filling with MTA; GIV-apical MTA plug + filling with MTA + metallic post (Reforpost I). A polyether impression material was used to simulate the periodontal ligament. The specimens were submitted to a compressive load at a crosshead speed of 0.5 mm min(-1) in a servo-hydraulic universal testing machine (MTS 810) applied at 45 degrees to the long axis of the tooth until failure. Data were submitted to statistical analysis by the Kruskal-Wallis test at 5% significance level.Results GIV presented the highest fracture resistance (32.7N) and differed significantly from the other groups (P < 0.05). No statistically difference was found between GII (16.6N) and GIII (23.4N) (P > 0.05). GIII had a significantly higher fracture resistance than GI (P < 0.05).Conclusions the use of MTA + metallic post as an intra-radicular reinforcement treatment increased the resistance to fracture of weakened bovine teeth in an experimental immature tooth model.

Experimental model for transanal endorectal pull-through surgery. Technique of de la Torre and Ortega

Background: We describe an experimental model for transanal endorectal pull-through surgery using the method of de]a Torre and Ortega that can be used for training purposes in experimental laboratories.Methods: Ten rabbits were submitted to the transanal endorectal pull-through technique of de la Torre and Ortega. Animals were randomly selected in the Botucatu School of Medicine experimental laboratory. Animals weighted between 2800 and 4400 g. Colons were not prepared, and antibiotic therapy was not used; dipyrone(1) was administered postoperatively for analgesic purposes. We standardized resected segment size, recorded surgical time, and observed Survival and possible complications for 1 month.Results: All animals survived the initial follow-up period without infection. Bowel movements returned quickly, and all animals were evacuating regularly within the first 24 hours. Mean surgical time was 48.6 minutes.Conclusions: the experimental model proposed in this study is very useful for training and improving surgical techniques using the method of de la Torre and Ortega. The rabbit is an excellent animal for this surgery because of its size and postoperative resistance. (c) 2005 Elsevier B.V. All rights reserved.

An attack signature model to computer security intrusion detection

Internal and external computer network attacks or security threats occur according to standards and follow a set of subsequent steps, allowing to establish profiles or patterns. This well-known behavior is the basis of signature analysis intrusion detection systems. This work presents a new attack signature model to be applied on network-based intrusion detection systems engines. The AISF (ACME! Intrusion Signature Format) model is built upon XML technology and works on intrusion signatures handling and analysis, from storage to manipulation. Using this new model, the process of storing and analyzing information about intrusion signatures for further use by an IDS become a less difficult and standardized process.

Open and standardized tools for smart transducer networking

IEEE 1451 Standard is intended to address the smart transducer interfacing problematic in network environments. Usually, proprietary hardware and software is a very efficient solution to in planent the IEEE 1451 normative, although can be expensive and inflexible. In contrast, the use of open and standardized tools for implementing the IEEE 1451 normative is proposed in this paper. Tools such as Java and Phyton programming languages, Linux, programmable logic technology, Personal Computer resources and Ethernet architecture were integrated in order to constructa network node based on the IEEE 1451 standards. The node can be applied in systems based on the client-server communication model The evaluation of the employed tools and expermental results are presented. © 2005 IEEE.

Galleria mellonella as a model host for human pathogens: recent studies and new perspectives

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

In Vivo Evaluation of the Phenox CRC Mechanical Thrombectomy Device in a Swine Model of Acute Vessel Occlusion

Mechanical thrombectomy in ischemic stroke is of increasing interest as it is a promising strategy for fast and efficient recanalization. Several thrombectomy devices have been introduced to the armentarium of mechanical thrombectomy. Currently, new devices are under development and are continuously added to the neurointerventional tool box. Each device advocated so far has a different design and mechanical properties in terms of thrombus-device interaction. Therefore, a systematic evaluation under standardized conditions in vivo of these new devices is needed. The purpose of this study was to evaluate the efficiency, thrombus-device interaction, and potential complications of the novel Phenox CRC for distal mechanical thrombectomy in vivo. The device was evaluated in an established animal model in the swine. Recanalization rate, thromboembolic events, vasospasm, and complications were assessed. Radiopaque thrombi (2 cm length) were used for the visualization of thrombus-device interaction during retrieval. The Phenox CRC (4 mm diameter) was assessed in 15 vessel occlusions. For every occlusion a maximum of 3 retrieval attempts were performed. Complete recanalization (TICI 3/TIMI 3) was achieved in 86.7% of vessel occlusions. In 66.7% (10/15), the first retrieval attempt was successful, and in 20% (3/15), the second attempt led to complete recanalization of the parent artery. In 2 cases (13.3%) thrombus retrieval was not successful (TICI 0/TIMI 0). In 1 case (6.7%) a minor embolic event occurred in a small side branch. No distal thromboembolic event was observed during the study. Thrombus-device interaction illustrated the entrapment of the thrombus by the microfilaments and the proximal cage of the device. No significant thrombus compression was observed. No vessel perforation, dissection, or fracture of the device occurred. In this small animal study, the Phenox CRC was a safe and effective device for mechanical thrombectomy. The unique design with a combination of microfilaments and proximal cage reduces thrombus compression with a consequently high recanalization and low complication rate.

The effect of rhBMP-2 around endosseous implants with and without membranes in the canine model

BACKGROUND: Bone morphogenetic protein (BMP) is a potent differentiating agent for cells of the osteoblastic lineage. It has been used in the oral cavity under a variety of indications and with different carriers. However, the optimal carrier for each indication is not known. This study examined a synthetic bioabsorbable carrier for BMP used in osseous defects around dental implants in the canine mandible. METHODS: Twelve canines had their mandibular four premolars and first molar teeth extracted bilaterally. After 5 months, four implants were placed with standardized circumferential defects around the coronal 4 mm of each implant. One-half of the defects received a polylactide/glycolide (PLGA) polymer carrier with or without recombinant human BMP-2 (rhBMP-2), and the other half received a collagen carrier with or without rhBMP-2. Additionally, one-half of the implants were covered with a non-resorbable (expanded polytetrafluoroethylene [ePTFE]) membrane to exclude soft tissues. Animals were sacrificed either 4 or 12 weeks later. Histomorphometric analysis included the percentage of new bone contact with the implant, the area of new bone, and the percentage of defect fill. This article describes results with the PLGA carrier. RESULTS: All implants demonstrated clinical and radiographic success with the amount of new bone formed dependent on the time and presence/absence of rhBMP-2 and presence/absence of a membrane. The percentage of bone-to-implant contact was greater with rhBMP-2, and after 12 weeks of healing, there was approximately one-third of the implant contacting bone in the defect site. After 4 weeks, the presence of a membrane appeared to slow new bone area formation. The percentage of fill in membrane-treated sites with rhBMP-2 rose from 24% fill to 42% after 4 and 12 weeks, respectively. Without rhBMP-2, the percentage of fill was 14% rising to 36% fill, respectively. CONCLUSIONS: After 4 weeks, the rhBMP-2-treated sites had a significantly higher percentage of contact, more new bone area, and higher percentage of defect fill than the sites without rhBMP-2. After 12 weeks, there was no significant difference in sites with or without rhBMP-2 regarding percentage of contact, new bone area, or percentage of defect fill. In regard to these three outcomes, comparing the results with this carrier to the results reported earlier with a collagen carrier in this study, only the area of new bone was significantly different with the collagen carrier resulting in greater bone than the PLGA carrier. Thus, the PLGA carrier for rhBMP-2 significantly stimulated bone formation around dental implants in this model after 1 month but not after 3 months of healing. The use of this growth factor and carrier combination appears to stimulate early bone healing events around the implants but not quite to the same degree as a collagen carrier.

Cyclooxygenase-2 inhibition selectively attenuates bone morphogenetic protein-6 synthesis and bone formation during guided tissue regeneration in a rat model

OBJECTIVES: Bone formation during guided tissue regeneration is a tightly regulated process involving cells, extracellular matrix and growth factors. The aims of this study were (i) to examine the expression of cyclooxygenase-2 (COX-2) during bone regeneration and (ii) the effects of selective COX-2 inhibition on osseous regeneration and growth factor expression in the rodent femur model. MATERIAL AND METHODS: A standardized transcortical defect of 5 x 1.5 mm was prepared in the femur of 12 male rats and a closed half-cylindrical titanium chamber was placed over the defect. The expression of COX-2 and of platelet-derived growth factor-B (PDGF-B), bone morphogenetic protein-6 (BMP-6) and insulin-like growth factor-I/II (IGF-I/II) was analyzed at Days 3, 7, 21 and 28 semiquantitatively by reverse transcriptase-polymerase chain reaction and immunohistochemistry. The effects of COX-2 inhibition by intraperitoneal injection of NS-398 (3 mg/kg/day) were analyzed in five additional animals sacrificed at Day 14. RESULTS: Histomorphometry revealed that new bone formation occurred in the cortical defect area as well as in the supracortical region, i.e. region within the chamber by Day 7 and increased through Day 28. Immunohistochemical evidence of COX-2 and PDGF-B levels were observed early (i.e. Day 3) and decreased rapidly by Day 7. BMP-6 expression was maximal at Day 3 and slowly declined by Day 28. In contrast, IGF-I/II expression gradually increased during the 28-day period. Systemic administration NS-398 caused a statistically significant reduction (P<0.05) in new bone formation (25-30%) and was associated with a statistically significant reduction in BMP-6 protein and mRNA expression (50% and 65% at P<0.05 and P<0.01, respectively). PDGF-B mRNA or protein expression was not affected by NS-398 treatment. CONCLUSION: COX-2 inhibition resulted in reduced BMP-6 expression and impaired osseous regeneration suggesting an important role for COX-2-induced signaling in BMP synthesis and new bone formation.

Topical caspofungin for treatment of keratitis caused by Candida albicans in a rabbit model

Candida albicans is the most frequent cause of fungal keratitis in temperate regions. Caspofungin has potent activity against Candida spp. in a variety of clinical settings. Little is known, however, about its activity against fungal keratitis. We compared the efficacy of topical caspofungin with that of topical amphotericin B (AMB) in a rabbit model of experimental keratomycosis. Keratitis was induced with a standardized inoculum of Candida albicans (SC 5314) placed on the debrided cornea. Twenty-four hours after infection, animals were randomly assigned to treatment with 0.15% caspofungin, 0.5% caspofungin, 0.15% AMB, and a saline control (n = 12 rabbits in each group). For the first 12 h, treatment was repeated every 30 min and, after a 12-h pause, was resumed at hourly intervals for another 12 h. The animals were examined and killed 12 h after administration of the last dose. Treatment effects were evaluated by clinical assessment, fungal culture, and histopathology. Drug treatment significantly reduced corneal fungal recovery from 3.78 log10 CFU in saline-treated animals to 2.97, 1.76, and 1.18 log10 CFU in animals treated with 0.15% caspofungin, 0.5% caspofungin, and 0.15% AMB, respectively. By histopathology, the mean hyphal density was significantly lower in the corneas of treated animals than in those of the controls; there was no difference in hyphal densities between the different treatment groups. The depth of corneal invasion was not significantly reduced by the antifungal treatments. By clinical assessment, keratitis progressed in animals treated with saline, whereas disease progression was inhibited by all drug treatment regimens. In our rabbit model, 0.5% caspofungin was as effective as 0.15% AMB for the topical treatment of Candida keratitis. The potential clinical efficacy of caspofungin awaits further investigation.

Double-lung transplantation in the rat: an acute, syngeneic in situ model

BACKGROUND. The high rate of reperfusion injury in clinical lung transplantation mandates significant improvements in lung preservation. Innovations should be validated using standardized and low-cost experimental models. METHODS. The model introduced here is analyzed by comparing global lung function after varying ischemic times (2, 4, 8, 16, and 24 hours). A rat double-lung block is flush-perfused, and the main pulmonary artery and left atrium are connected to the left pulmonary artery and vein of a syngeneic recipient using a T-shaped stent. With pressure side ports and incorporated flow crystals, measurement of vascular resistance and graft oxygenation can be performed. The transplant is ventilated separately, and compliance and resistance are determined. RESULTS. The increase in the ischemic interval from 2 to 24 hours caused an increase in the alveolar arterial oxygen difference from 220 +/- 20 to 600 +/- 34 mm Hg, pulmonary vascular resistance from 198 +/- 76 to 638 +/- 212 mm Hg.mL-1.min-1, and resistance to airflow from 274 +/- 50 to 712 +/- 30 cm H2O/L H2O, and a decrease in pulmonary compliance from 0.4 +/- 0.05 to 0.12 +/- 0.06 mL/cm H2O. CONCLUSIONS. This in situ, syngeneic rat lung transplantation model offers an alternative to large animal models for verification of lung preservation solutions and for modification of donor or recipient treatment regimens.