A double antibody sandwich ELISA for rapid diagnosis of virus infection and to measure the humoral response against infectious bursal disease on clinical material

A double antibody sandwich ELISA (DAS-ELISA) was developed and employed for simultaneous direct detection of infectious bursal disease virus (IBDV) from bursal samples and to measure the humoral response, using the same basic immunoreagents, the purified and non-purified antigen, capture antibody and chicken hyperimmune sera were prepared, and standardized for this purpose, the DAS-ELISA was applied to both 80 bursal suspensions and 224 corresponding serum samples from vaccinated and non-vaccinated commercial hocks, Bursae samples were collected at 2 weeks of age, and submitted to histological examination, virus isolation in specific pathogen-free chickens embryos, and the DAS-ELISA technique, Serum titres obtained in indirect ELISA and serum neutralization test were compared with those in DAS-ELISA, the agreement was 80% between DAS-ELISA, and the conventional techniques, with high sensitivity (87%) and specificity (90%).

Chicken embryo related (CER) cell line for quantification of rabies neutralizing antibody by fluorescent focus inhibition test

Levels of rabies virus neutralization antibody in sera from vaccinated dogs and cattle were either measured by mouse neutralization test (MNT) or by rapid fluorescent focus inhibition test (RFFIT), performed on CER monolayers. The two tests were compared for their ability to detect the 0.5 International Units/ml (I.U.) recommended by the World Health Organization (WHO) as the minimum response for proof of rabies immunization. A significant correlation was found between the two tests (n = 211; r = 0.9949 in dogs and 0.9307 in cows, p < 0.001), good sensitivity (87.5%), specificity (94.7%) and agreement (96.6%) as well. RFFIT method standardized on CER cell system for neutralizing antibodies detection turns the diagnosis easier and less expensive, specially when a great number of samples must be tested from endemic areas as commonly found in Brazil. (c) 2005 the International Association for Biologicals. Published by Elsevier Ltd. All rights reserved.

Comparison between the counter immunoelectrophoresis test and mouse neutralization test for the detection of antibodies against rabies virus in dog sera

The detection of rabies antibodies is extremely valuable for epidemiological studies, determination of immune status in man, animals, and for the diagnosis of the disease. Several serological procedures have been described for this purpose. The present study reports a comparison between counterimmunoelectrophoresis test (CIET) and mouse neutralization test (MNT) in the detection of antibodies against rabies virus from 212 serum samples of vaccinated dogs. The agreement between both techniques was 79.7% and a significative association was demonstrated. The correlation coefficients between MNT and the CIET titers was determined considering 88 samples showing positive results in both techniques [CIET = 2 and MNT = 5 (0.13 IU/ml)] and resulted r² = 0.7926 (p < 0.001). The performance of CIET system was technically simple, cheap and rapid, and thereby it could be useful for serological monitoring of dog vaccination campaigns as well as for individual analysis.

Soft tissue profile changes after rapid maxillary expansion with a bonded expander

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

Development of a rapid one-step immunochromatographic assay for HCV core antigen detection

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Animal model of rapid crystalloid infusion in rats

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

Influence of culture media in viability test of conidia of entomopathogenic fungi

O presente trabalho objetivou investigar se meios de cultura utilizados em teste de viabilidade afetam a germinação de conídios de cinco isolados de Lecanicillium lecanii, cinco de Beauveria bassiana e quatro de Paecilomyces fumosoroseus. Os testes foram realizados em lâminas de microscopia contendo um dos seguintes meios de cultura: Ágar-água (AA), Meio Mínimo (MM), Batata, dextrose e ágar (BDA), Batata, dextrose, ágar e 1% de extrato de levedura (BDAL), Sabouraud, dextrose, ágar e extrato de levedura (SDAL) e Meio Completo (MC). Delimitaram-se três áreas por lâmina e em cada uma aplicou-se 0,05mL de uma suspensão com concentração de 5,5 x 105 conídios ml-1. Para cada isolado foi realizado um bioensaio, com seis tratamentos e cinco repetições. Avaliou-se a germinação 15 horas após incubação, a 26±0,5ºC. Os meios de cultura influíram na capacidade de germinação das três espécies estudadas, ocorrendo variações inter e intraespecíficas. Verificou-se que os meios Completo e BDA proporcionaram as maiores porcentagens de germinação dos isolados de L. lecanii, sendo que e as menores foram obtidas nos meios SDAL e AA. Os meios ricos em nutrientes (BDA, BDAL e Completo) favoreceram a germinação dos isolados de B. bassiana, o que não ocorreu com os meios pobres (AA e MM). Nos meios Completo e BDA foram obtidas as maiores porcentagens de germinação dos isolados de P. fumosoroseus. As menores percentagens, por sua vez, foram obtidas no meio SDAL. Entretanto, alguns isolados apresentaram alta germinação em meios pobres em nutrientes (AA e MM).

Evaluation of a novel kit (TF-test) for the diagnosis of intestinal parasitic infections

Intestinal parasitic infections are currently a source of concern for Public Health agencies in developing and developed countries. Since three ovum-and-parasite stool examinations have been demonstrated to provide sensitive results, we designed a practical and economical kit (TF-Test) that is now commercially available (Immunoassay Com. Ind. Ltda., S (a) over tildeo Paulo, Brazil). This kit allows the separate collection of three fecal specimens into a preservative solution. The specimens are then pooled, double-filtered, and concentrated by a single rapid centrifugation process. The TF-Test was evaluated in four different laboratories in a study using 1,102 outpatients and individuals living in an endemic area for enteroparasitosis. The overall sensitivity found using the TF-Test (86.2-97.8%) was significantly higher (P<0.01) than the sensitivity of conventional techniques such as the Coprotest (NIL Comercio Exterior Ltda, São Paulo, Brazil) and the combination of Lutz/Hoffman, Faust, and Rugai techniques (De Carli, Diagnostico Laboratorial das Parasitoses Humanas. Metodos e Tecnicas, 1994), which ranged from 48.3% to 75.9%. When the above combined three specimen technique was repeated with three specimens collected on different days, its sensitivity became similar (P > 0.01) to that of the TF-Test. The kappa index values of agreement for the TF-Test were consistent (P < 0.01), being higher and ranking in a better position than conventional techniques. The high sensitivity, cost/benefit ratio, and practical aspects demonstrate that the TF-Test is suitable for individual diagnosis, epidemiological inquiries, or evaluation of chemotherapy in treated communities. (C) 2004 Wiley-Liss, Inc.

Comparison of two immunochromatographic assays and the indirect immunofluorscence antibody test for diagnosis of Trypanosoma cruzi infection in dogs in south central Louisiana

Two rapid tests evaluated in dogs considered to be of high risk of Infection with the Chagas parasite Trypanosoma cruzi using two immunochromatographic assays. Trypanosoma Detect (TM) for canine, InBios, Seattle, WA and CHAGAS STAT-PAK (TM) assay, Chembio Diagnostic Systems, Medford, NY, in south central Louisiana. For this purpose a serological survey was carried out in a total of 122 dogs and a serum bank was created. These 122 animals were first tested by IFAT that was used as the standard test From the serum bank 50 samples were tested using the two rapid Chagas assays and results compared to the standard test IFAT The serological survey using IFAT showed it prevalence of T cruzi infection in 22.1% of the tested dogs. In the immunochromatographic assays. 13 and 11 animals were positive on rapid assay Trypanosoma Detect (TM) for canine, InBios and CHAGAS STAT-PAK (TM), Chembio Diagnostic Systems, respectively compared to 11 positive by IFAT. These two immunochromatographic tests have shown high susceptibility and specificity compared to our standard method IFAT. The rapid, easy and accurate screening assays used in conjunction with confirmatory tests, would be an excellent tool for veterinarians to diagnose T cruzi infection. Early detection of T cruzi infection may prevent complications through an effective treatment. Greater awareness by veterinarians of the risk. clinical findings, history along with diagnostic methods will contribute greatly to an understanding of the true prevalence of Chagas disease in dogs in Louisiana. (c) 2009 Elsevier B.V. All rights reserved.

Determination of methyldopa in pharmaceutical formulations by combined spot test-diffuse reflectance spectroscopy

This paper describes a very simple and rapid quantitative reflectance spot test procedure for the determination of methyldopa in pharmaceutical formulations. This method is based on the complexation reaction of methyldopa with molybdate ions yielding a yellow stable complex on filter paper. Reflectance measurements were carried out at 410 nm. Under optimal conditions, the calibration graphs obtained for methyldopa by plotting the optical density of the reflectance signal (A R) vs. the log of the concentration were linear from 6.30 × 10 -3 to 1.89 × 10 -2 mol L -1, with a correlation coefficient of 0.998. The detection limit was 2.74 × 10 -3 mol L -1 (R.S.D. = 1.02%) for methyldopa. The common excipients used as additives in pharmaceuticals do not interfere in the proposed method. The method was applied to determine metyldopa in commercial pharmaceutical formulations. The results obtained by the proposed method compare favorably with those obtained by an official procedure at 95% confidence level. ©2006 Sociedade Brasileira de Química.

Bond strength of hard chairside reline resins to a rapid polymerizing denture base resin before and after thermal cycling

Purpose: This study assessed the shear bond strength of 4 hard chairside reline resins (Kooliner, Tokuso Rebase Fast, Duraliner II, Ufi Gel Hard) to a rapid polymerizing denture base resin (QC-20) processed using 2 polymerization cycles (A or B), before and after thermal cycling. Materials and Methods: Cylinders (3.5 mm x 5.0 mm) of the reline resins were bonded to cylinders of QC-20 polymerized using cycle A (boiling water-20 minutes) or B (boiling water; remove heat-20 minutes; boiling water-20 minutes). For each reline resin/polymerization cycle combination, 10 specimens (groups CAt e CBt) were thermally cycled (5 and 55°C; dwell time 30 seconds; 2,000 cycles); the other 10 were tested without thermal cycling (groups CAwt ad CBwt). Shear bond tests (0.5 mm/min) were performed on the specimens and the failure mode was assessed. Data were analyzed by 3-way ANOVA and Newman-Keuls post-hoc test (α=.05). Results: QC-20 resin demonstrated the lowest bond strengths among the reline materials (P<.05) and mainly failed cohesively. Overall, the bond strength of the hard chairside reline resins were similar (10.09±1.40 to 15.17±1.73 MPa) and most of the failures were adhesive/cohesive (mixed mode). However, Ufi Gel Hard bonded to QC-20 polymerized using cycle A and not thermally cycled showed the highest bond strength (P<.001). When Tokuso Rebase Fast and Duraliner II were bonded to QC-20 resin polymerized using cycle A, the bond strength was increased (P=.043) after thermal cycling. Conclusions: QC-20 displayed the lowest bond strength values in all groups. In general, the bond strengths of the hard chairside reline resins were comparable and not affected by polymerization cycle of QC-20 resin and thermal cycling.

A simplified reflectometric method for the rapid determination of dipyrone in pharmaceutical formulations

This paper describes a simple, portable and environmentally friendly method for the rapid determination of dipyrone in pharmaceuticals by using diffuse reflectance spectroscopy. The proposed method is based on the reflectance measurements of the orange compound produced from the spot test reaction between dipyrone and p-dimethylaminocinnamaldehyde (p-DAC), in acid medium, using a filter paper as solid support. Experimental design methodologies were used to optimize the measurement conditions. All reflectance measurements were carried out at 510 nm and the linear range was from 1.42 × 10-4-2.85 × 10-3 mol L-1, with a correlation coefficient of 0.999. The limit of detection (LOD) and the limit of quantification (LOQ) were 1.20 × 10-5 mol L-1 and 4.00 × 10-5 mol L-1, respectively. The intraday precision and interday precision were studied for 10 replicate analyses of 7.90 × 10-4 mol L-1 dipyrone solution. The coefficients of variation were 1.1 and 0.9%, respectively. The proposed method was applied successfully to the determination of dipyrone in commercial brands of pharmaceuticals. No interferences were observed from the common excipients in formulations. The results obtained by the proposed method were favorably compared with those given by the Brazilian Pharmacopoeia procedure at 95% confidence level. ©2007 Sociedade Brasileira de Química.

Fatigue and rapid hamstring/quadriceps force capacity in professional soccer players

The aim of this study was to investigate the effect of fatigue induced by an exhaustive laboratory-based soccer-specific exercise on different hamstrings/quadriceps (H:Q) ratios of soccer players. Twenty-two male professional soccer players (23·1 ± 3·4 year) performed maximal eccentric (ecc) and concentric (con) contractions for knee extensors (KE) and flexors (KF) at 60° s-1 and 180° s-1 to assess conventional (Hcon:Qcon) and functional (Hecc:Qcon) ratios. Additionally, they performed maximal voluntary isometric contraction for KE and KF, from which the maximal muscle strength, rate of force development (RFD) and RFD H:Q strength ratio (RFDH:Q) were extracted. Thereafter, subjects were performed an exhaustive laboratory-based soccer-specific exercise and a posttest similar to the pretest. There was significant reduction in Hcon:Qcon (0·60 ± 0·06 versus 0·58 ± 0·06, P<0·05) and in Hecc:Qcon (1·29 ± 0·2 versus 1·16 ± 0·2, P<0·01) after the soccer-specific exercise. However, no significant difference between Pre and Post exercise conditions was found for RFDH:Q at 0-50 (0·53 ± 0·23 versus 0·57 ± 0·24, P>0·05) and 0-100 ms (0·53 ± 0·17 versus 0·55 ± 0·17, P>0·05). In conclusion, H:Q strength ratios based on peak force values are more affected by fatigue than RFDH:Q obtained during early contraction phase. Thus, fatigue induced by soccer-specific intermittent protocol seems not reduce the potential for knee joint stabilization during the initial phase of voluntary muscle contraction. copy; 2012 Scandinavian Society of Clinical Physiology and Nuclear Medicine.

Cebus cf. apella exhibits rapid acquisition of complex stimulus relations and emergent performance by exclusion

A "second generation" matching-to-sample procedure that minimizes past sources of artifacts involves (1) successive discrimination between sample stimuli, (2) stimulus displays ranging from four to 16 comparisons, (3) variable stimulus locations to avoid unwanted stimulus-location control, and (4) high accuracy levels (e.g., 90% correct on a 16-choice task in which chance accuracy is 6%). Examples of behavioral engineering with experienced capuchin monkeys included four-choice matching problems with video images of monkeys with substantially above-chance matching in a single session and 90% matching within six sessions. Exclusion performance was demonstrated by interspersing non-identical sample-comparison pairs within a baseline of a nine-comparison identity-matching-to-sample procedure with pictures as stimuli. The test for exclusion presented the newly "mapped" stimulus in a situation in which exclusion was not possible. Degradation of matching between physically non-identical forms occurred while baseline identity accuracy was sustained at high levels, thus confirming that Cebus cf. apella is capable of exclusion. Additionally, exclusion performance when baseline matching relations involved non-identical stimuli was shown.

A test of the utility of DNA barcoding in the radiation of the freshwater stingray genus Potamotrygon (Potamotrygonidae, Myliobatiformes)

DNA barcoding is a recently proposed global standard in taxonomy based on DNA sequences. The two main goals of DNA barcoding methodology are assignment of specimens to a species and discovery of new species. There are two main underlying assumptions: i) reciprocal monophyly of species, and ii) intraspecific divergence is always less than interspecific divergence. Here we present a phylogenetic analysis of the family Potamotrygonidae based on mitochondrial cytochrome c oxidase I gene, sampling 10 out of the 18 to 20 valid species including two non-described species. Potamotrygonidae systematics is still not fully resolved with several still-to-be-described species while some other species are difficult to delimit due to overlap in morphological characters and because of sharing a complex color patterns. Our results suggest that the family passed through a process of rapid speciation and that the species Potamotrygon motoro, P. scobina, and P. orbignyi share haplotypes extensively. Our results suggest that systems of identification of specimens based on DNA sequences, together with morphological and/or ecological characters, can aid taxonomic studies, but delimitation of new species based on threshold values of genetic distances are overly simplistic and misleading.