yeda nogueira - Academia.edu (original) (raw)
Papers by yeda nogueira
Revista do Instituto Adolfo Lutz, Dec 30, 1992
Revista do Instituto Adolfo Lutz, Jun 28, 1996
During this work Sokol's classical method, slightly modified, was used in order to obtain lhe pur... more During this work Sokol's classical method, slightly modified, was used in order to obtain lhe purified fraction of rabies virus, A filtration in Sephadex G-50 column took place followed by another filtration in Sephadex G-75, eliminating the enzimatic treatment with DNase and RNase. The elution in Sephadex G-50 separated two peaks with activity in 280nm (Absorbance). Peak I has separed a bovine albumine serum and Peak II has separated rnost of the viral fraction purified, free of such eontaminats as serum or eellular proteins. Peak I was submitted to a saecharose gradient clumping ali viral particles components and they were bioehemically eharaeterized throughout the electrophoretic separation in poliaerylamide gel SDS-PAGE and immunogcnically throughout crossed immunoelectrophoresis, Crossed immunoelectrophoresis was used to follow all the purification process, allowing the observation of which fraetions were eliminated in each step of the processo On the other hand, two advantages could be obtained when the bovine albumine serum was eliminated in second filtration through the Sephadex G-50 and when erossed immunocletrophorcsis technique allowed a complete peer-view of the processo This aproach is very interesting when a large scale produetion process of purified antigen is involved for it allows the control of the purity degree and the attainment of a good quality control of the product.
The McCoy cell line has showed high sensivity to rabies virus. This cell line presents cythopatic... more The McCoy cell line has showed high sensivity to rabies virus. This cell line presents cythopatic effect (CPE), when infected with rabies virus. Three strains of rabies virus were tested: CVS, ERA, and PVI and three street virus were isolated: one from a patient bitten by bat, other from a rabid dog and another from bovine. This cell line was used for virus titration purposes due to the presence of cytophatic effect, two techniques for observation of the CPE were used and were compared to the classical method of intracerebral inoculation in mice
Brazilian Journal of Veterinary Research and Animal Science, 2003
Evaluation of the performance of the modified direct agglutination test (MAT) for detection of To... more Evaluation of the performance of the modified direct agglutination test (MAT) for detection of Toxoplasma gondii antibodies in dogs Avaliação da performance do teste de aglutinação modifica (MAT) paa a detecção de anticorpos anti-Toxoplasma gondii em cães
Rev Inst Adolfo Lutz, 1992
Revista do Instituto de Medicina Tropical de São Paulo
Leishmanias can be produced by inoculation in conditioned McCoy cell culture growth medium (CGM).... more Leishmanias can be produced by inoculation in conditioned McCoy cell culture growth medium (CGM). Leishmania (Leishmania) infantum chagasi (100 parasites) grown in NNN medium was inoculated in 2.5 mL CGM, kept in plates (24 wells) and its multiplication was observed for five days (120 hours). After day 5, the medium was saturated with the flagellate forms of the parasite (promastigotes). The reproduction of the leishmanias was observed every 24 hours and the number of parasites was calculated by counting the parasites in a drop of 10 microL and photomicrographed. So the number of Leishmanias was adjusted to 1 mL volume. The advantage of the technique by isolation of Leishmania in CGM demonstrated in this study is its low cost and high efficacy even with a small quantity of parasites (10(2) promastigotes) used as inoculum. Additionally, isolation of the leishmania can be obtained together with an increase in their density (180 times) as observed by growth kinetics, within a shorter t...
Revista de saúde pública, 2004
No population-based studies have been conducted to show the potential for the use of virological ... more No population-based studies have been conducted to show the potential for the use of virological diagnosis of the rabies virus. The objective of the present study was to estimate accuracy parameters for the isolation of the rabies virus in McCoy cells as an alternative method and to compare this with the use of murine neuroblastoma (N2A) cells, which is considered to be a reference method. An evaluation was performed on 120 bats collected at random in the Atlantic Forest of the State of São Paulo. The immunofluorescence reaction was utilized for the detection of the rabies virus isolated from the brain of these bats and the presence of the virus was tested in the two cell culture systems. Two data sets were constructed with the results and the analysis was performed using the computer methods for diagnosis tests (CMDT) software by means of the two-graph receiver operating characteristic (TG-ROC) technique to determine sensitivity and specificity parameters, as well as other indicato...
Revista do Instituto de Medicina Tropical de São Paulo, 2007
The Western blot technique was used to demonstrate the presence of antibodies in the blood of dog... more The Western blot technique was used to demonstrate the presence of antibodies in the blood of dogs that presented canine visceral leishmaniasis. This technique was used against some specific molecules present in the lysate of the promastigote form of Leshmania chagasi.Through the association of the results of the Western blot technique with the morphological alterations seen as a result of the serum neutralization technique performed in McCoy cells (which mimetizes the macrophage) it was possible to observe the role of some molecules of great relevance in determining the disease in symptomatic dogs as well as that of some other molecules associated with asymptomatic infected dogs that may become transmitters as well as differentiating them as asymptomatic resistant dogs. In the sera analyses carried out during the immunobloting a variation of 9 to 27 immunoreacting bands was observed, which were then compared using Dice's similarity coefficient. In the dendrogram constructed on ...
Revista de saúde pública, 2009
To analyze the accuracy of the diagnosis of two protocols of indirect immunofluorescence assays f... more To analyze the accuracy of the diagnosis of two protocols of indirect immunofluorescence assays for canine visceral leishmaniasis. Dogs from the seroepidemiological survey conducted in an endemic area of the cities of Araçatuba and Andradina, in Northwestern São Paulo state, in 2003, and in a non-endemic area of the metropolitan region of São Paulo, were used to assess two protocols of indirect immunofluorescence assay (IFA) for leishmaniasis: one using a Leishmania major heterologous antigen (IFA-BM) and another using a Leishmania chagasi homologous antigen (IFA-CH). Two-graph receiver operating characteristic (TG-ROC) analysis was used to estimate accuracy. TG-ROC analysis compared 1:20 dilution readings of the homologous antigen (IFA-CH), considered as reference test, with IFA-BM dilutions (heterologous antigen). The 1:20 dilution used in the IFA-CH test showed the best contingency coefficient (0.755) and the highest strength of association between the two variables studied (chi-...
Brazilian Journal of …, 2003
Toxoplasmose é uma zoonose que vem sendo objeto de estudos no Brasil e em todas as partes do mund... more Toxoplasmose é uma zoonose que vem sendo objeto de estudos no Brasil e em todas as partes do mundo. Os cães são considerados sentinelas da infecção, podendo carrear o agente pelo ambiente. Levantamentos soro-epidemiológicos desses animais são importantes ferramentas de vigilância e controle da doença em programas de saúde. Neste estudo a performance do Teste de Aglutinação Modificada (MAT) no sorodiagnóstico da toxoplasmose canina foi avaliado e comparado à reação de imunoflorescência indireta (RIFI). Uma amostra de 157 soros de cães do município de Monte Negro, Rondônia, com 76.40% de animais positivos ao Toxoplasma gondii (RIFI =16) foi analisado utilizando o MAT (=25) e apresentou sensibilidade de 85,00% (Intervalo de Confiança 95,00%: 79,4-90,60%) e especificidade de 100,00%.
Revista do Instituto de Medicina Tropical de São Paulo
The Western blot technique was used to demonstrate the presence of antibodies in the blood of dog... more The Western blot technique was used to demonstrate the presence of antibodies in the blood of dogs that presented canine visceral leishmaniasis. This technique was used against some specific molecules present in the lysate of the promastigote form of Leshmania chagasi. Through the association of the results of the Western blot technique with the morphological alterations seen as a result of the serum neutralization technique performed in McCoy cells (which mimetizes the macrophage) it was possible to observe the role of some molecules of great relevance in determining the disease in symptomatic dogs as well as that of some other molecules associated with asymptomatic infected dogs that may become transmitters as well as differentiating them as asymptomatic resistant dogs. In the sera analyses carried out during the immunobloting a variation of 9 to 27 immunoreacting bands was observed, which were then compared using Dice's similarity coefficient. In the dendrogram constructed on the basis of the coefficient, 50% similarity was observed among the total number of reagent bands with the promastigote lysate, thus creating five groups. The main difference observed related to the clinical condition of the dogs: symptomatic and asymptomatic dogs were found in separate groups. The asymptomatic group of dogs was distributed in two different places in the dendrogram because they presented two different behavior patterns regarding the cellular morphology in the serum neutralization reaction: the presence or absence of cellular lysis. According to this analysis it is possible to evaluate the immune status and associate it with specific markers observed in the reaction found in the Western blot strips.
Revista de Saúde Pública, 2009
ABSTRACT OBJECTIVE: To analyze the accuracy of the diagnosis of two protocols of indirect immunof... more ABSTRACT OBJECTIVE: To analyze the accuracy of the diagnosis of two protocols of indirect immunofluorescence assays for canine visceral leishmaniasis.
Revista do Instituto de Medicina Tropical de São Paulo
Several studies have recently shown the use of recombinant rabies virus as potential vector-viral... more Several studies have recently shown the use of recombinant rabies virus as potential vector-viral vaccine for HIV-1. The sequence homology between gp 120 and rabies virus glycoprotein has been reported. The McCoy cell line has therefore been used to show CD4+ or CD4+ like receptors. Samples of HIV-1 were isolated, when plasma of HIV-1 positive patients was inoculated in the McCoy cell line. The virus infection was then studied during successive virus passages. The proteins released in the extra cellular medium were checked for protein activity, by exposure to SDS Electrophoresis and blotting to nitro-cellulose filter, then reacting with sera of HIV positive and negative patients. Successive passages were performed, and showed viral replication, membrane permeabilization, the syncytium formation, and the cellular lysis (cytopathic effect). Flow cytometry analysis shows clear evidence that CD4+ receptors are present in this cell line, which enhances the likelihood of easy isolation and replication of HIV. The results observed allow the use of this cell line as a possible model for isolating HIV, as well as for carrying out studies of the dynamics of viral infection in several situations, including exposure to drugs in pharmacological studies, and possibly studies and analyses of the immune response in vaccine therapies.
Revista do Instituto de Medicina Tropical de São …, 1998
We evaluated the components of the Fuenzalida-Palacios antirabies vaccine, which is till used in ... more We evaluated the components of the Fuenzalida-Palacios antirabies vaccine, which is till used in most developing countries in human immunization for treatment and prophylaxis. This vaccine is prepared from newborn mouse brains at 1% concentration. Even though the vaccine is considered to have a low myelin content, it is not fully free of myelin or of other undesirable components that might trigger adverse effects after vaccination. The most severe effect is a post-vaccination neuroparalytic accident associated with Guillain-Barré syndrome. In the present study we demonstrate how the vaccines produced and distributed by different laboratories show different component patterns with different degrees of impurity and with varying protein concentrations, indicating that production processes can vary from one laboratory to another. These differences, which could be resolved using a better quality control process, may affect and impair immunization, with consequent risks and adverse effects after vaccination. We used crossed immunoelectrophoresis to evaluate and demonstrate the possibility of quality control in vaccine production, reducing the risk factors possibly involved in these immunizing products.
Revista de Saúde Pública, 2004
No population-based studies have been conducted to show the potential for the use of virological ... more No population-based studies have been conducted to show the potential for the use of virological diagnosis of the rabies virus. The objective of the present study was to estimate accuracy parameters for the isolation of the rabies virus in McCoy cells as an alternative method and to compare this with the use of murine neuroblastoma (N2A) cells, which is considered to be a reference method.
Revista do Instituto Adolfo Lutz, Dec 30, 1992
Revista do Instituto Adolfo Lutz, Jun 28, 1996
During this work Sokol's classical method, slightly modified, was used in order to obtain lhe pur... more During this work Sokol's classical method, slightly modified, was used in order to obtain lhe purified fraction of rabies virus, A filtration in Sephadex G-50 column took place followed by another filtration in Sephadex G-75, eliminating the enzimatic treatment with DNase and RNase. The elution in Sephadex G-50 separated two peaks with activity in 280nm (Absorbance). Peak I has separed a bovine albumine serum and Peak II has separated rnost of the viral fraction purified, free of such eontaminats as serum or eellular proteins. Peak I was submitted to a saecharose gradient clumping ali viral particles components and they were bioehemically eharaeterized throughout the electrophoretic separation in poliaerylamide gel SDS-PAGE and immunogcnically throughout crossed immunoelectrophoresis, Crossed immunoelectrophoresis was used to follow all the purification process, allowing the observation of which fraetions were eliminated in each step of the processo On the other hand, two advantages could be obtained when the bovine albumine serum was eliminated in second filtration through the Sephadex G-50 and when erossed immunocletrophorcsis technique allowed a complete peer-view of the processo This aproach is very interesting when a large scale produetion process of purified antigen is involved for it allows the control of the purity degree and the attainment of a good quality control of the product.
The McCoy cell line has showed high sensivity to rabies virus. This cell line presents cythopatic... more The McCoy cell line has showed high sensivity to rabies virus. This cell line presents cythopatic effect (CPE), when infected with rabies virus. Three strains of rabies virus were tested: CVS, ERA, and PVI and three street virus were isolated: one from a patient bitten by bat, other from a rabid dog and another from bovine. This cell line was used for virus titration purposes due to the presence of cytophatic effect, two techniques for observation of the CPE were used and were compared to the classical method of intracerebral inoculation in mice
Brazilian Journal of Veterinary Research and Animal Science, 2003
Evaluation of the performance of the modified direct agglutination test (MAT) for detection of To... more Evaluation of the performance of the modified direct agglutination test (MAT) for detection of Toxoplasma gondii antibodies in dogs Avaliação da performance do teste de aglutinação modifica (MAT) paa a detecção de anticorpos anti-Toxoplasma gondii em cães
Rev Inst Adolfo Lutz, 1992
Revista do Instituto de Medicina Tropical de São Paulo
Leishmanias can be produced by inoculation in conditioned McCoy cell culture growth medium (CGM).... more Leishmanias can be produced by inoculation in conditioned McCoy cell culture growth medium (CGM). Leishmania (Leishmania) infantum chagasi (100 parasites) grown in NNN medium was inoculated in 2.5 mL CGM, kept in plates (24 wells) and its multiplication was observed for five days (120 hours). After day 5, the medium was saturated with the flagellate forms of the parasite (promastigotes). The reproduction of the leishmanias was observed every 24 hours and the number of parasites was calculated by counting the parasites in a drop of 10 microL and photomicrographed. So the number of Leishmanias was adjusted to 1 mL volume. The advantage of the technique by isolation of Leishmania in CGM demonstrated in this study is its low cost and high efficacy even with a small quantity of parasites (10(2) promastigotes) used as inoculum. Additionally, isolation of the leishmania can be obtained together with an increase in their density (180 times) as observed by growth kinetics, within a shorter t...
Revista de saúde pública, 2004
No population-based studies have been conducted to show the potential for the use of virological ... more No population-based studies have been conducted to show the potential for the use of virological diagnosis of the rabies virus. The objective of the present study was to estimate accuracy parameters for the isolation of the rabies virus in McCoy cells as an alternative method and to compare this with the use of murine neuroblastoma (N2A) cells, which is considered to be a reference method. An evaluation was performed on 120 bats collected at random in the Atlantic Forest of the State of São Paulo. The immunofluorescence reaction was utilized for the detection of the rabies virus isolated from the brain of these bats and the presence of the virus was tested in the two cell culture systems. Two data sets were constructed with the results and the analysis was performed using the computer methods for diagnosis tests (CMDT) software by means of the two-graph receiver operating characteristic (TG-ROC) technique to determine sensitivity and specificity parameters, as well as other indicato...
Revista do Instituto de Medicina Tropical de São Paulo, 2007
The Western blot technique was used to demonstrate the presence of antibodies in the blood of dog... more The Western blot technique was used to demonstrate the presence of antibodies in the blood of dogs that presented canine visceral leishmaniasis. This technique was used against some specific molecules present in the lysate of the promastigote form of Leshmania chagasi.Through the association of the results of the Western blot technique with the morphological alterations seen as a result of the serum neutralization technique performed in McCoy cells (which mimetizes the macrophage) it was possible to observe the role of some molecules of great relevance in determining the disease in symptomatic dogs as well as that of some other molecules associated with asymptomatic infected dogs that may become transmitters as well as differentiating them as asymptomatic resistant dogs. In the sera analyses carried out during the immunobloting a variation of 9 to 27 immunoreacting bands was observed, which were then compared using Dice's similarity coefficient. In the dendrogram constructed on ...
Revista de saúde pública, 2009
To analyze the accuracy of the diagnosis of two protocols of indirect immunofluorescence assays f... more To analyze the accuracy of the diagnosis of two protocols of indirect immunofluorescence assays for canine visceral leishmaniasis. Dogs from the seroepidemiological survey conducted in an endemic area of the cities of Araçatuba and Andradina, in Northwestern São Paulo state, in 2003, and in a non-endemic area of the metropolitan region of São Paulo, were used to assess two protocols of indirect immunofluorescence assay (IFA) for leishmaniasis: one using a Leishmania major heterologous antigen (IFA-BM) and another using a Leishmania chagasi homologous antigen (IFA-CH). Two-graph receiver operating characteristic (TG-ROC) analysis was used to estimate accuracy. TG-ROC analysis compared 1:20 dilution readings of the homologous antigen (IFA-CH), considered as reference test, with IFA-BM dilutions (heterologous antigen). The 1:20 dilution used in the IFA-CH test showed the best contingency coefficient (0.755) and the highest strength of association between the two variables studied (chi-...
Brazilian Journal of …, 2003
Toxoplasmose é uma zoonose que vem sendo objeto de estudos no Brasil e em todas as partes do mund... more Toxoplasmose é uma zoonose que vem sendo objeto de estudos no Brasil e em todas as partes do mundo. Os cães são considerados sentinelas da infecção, podendo carrear o agente pelo ambiente. Levantamentos soro-epidemiológicos desses animais são importantes ferramentas de vigilância e controle da doença em programas de saúde. Neste estudo a performance do Teste de Aglutinação Modificada (MAT) no sorodiagnóstico da toxoplasmose canina foi avaliado e comparado à reação de imunoflorescência indireta (RIFI). Uma amostra de 157 soros de cães do município de Monte Negro, Rondônia, com 76.40% de animais positivos ao Toxoplasma gondii (RIFI =16) foi analisado utilizando o MAT (=25) e apresentou sensibilidade de 85,00% (Intervalo de Confiança 95,00%: 79,4-90,60%) e especificidade de 100,00%.
Revista do Instituto de Medicina Tropical de São Paulo
The Western blot technique was used to demonstrate the presence of antibodies in the blood of dog... more The Western blot technique was used to demonstrate the presence of antibodies in the blood of dogs that presented canine visceral leishmaniasis. This technique was used against some specific molecules present in the lysate of the promastigote form of Leshmania chagasi. Through the association of the results of the Western blot technique with the morphological alterations seen as a result of the serum neutralization technique performed in McCoy cells (which mimetizes the macrophage) it was possible to observe the role of some molecules of great relevance in determining the disease in symptomatic dogs as well as that of some other molecules associated with asymptomatic infected dogs that may become transmitters as well as differentiating them as asymptomatic resistant dogs. In the sera analyses carried out during the immunobloting a variation of 9 to 27 immunoreacting bands was observed, which were then compared using Dice's similarity coefficient. In the dendrogram constructed on the basis of the coefficient, 50% similarity was observed among the total number of reagent bands with the promastigote lysate, thus creating five groups. The main difference observed related to the clinical condition of the dogs: symptomatic and asymptomatic dogs were found in separate groups. The asymptomatic group of dogs was distributed in two different places in the dendrogram because they presented two different behavior patterns regarding the cellular morphology in the serum neutralization reaction: the presence or absence of cellular lysis. According to this analysis it is possible to evaluate the immune status and associate it with specific markers observed in the reaction found in the Western blot strips.
Revista de Saúde Pública, 2009
ABSTRACT OBJECTIVE: To analyze the accuracy of the diagnosis of two protocols of indirect immunof... more ABSTRACT OBJECTIVE: To analyze the accuracy of the diagnosis of two protocols of indirect immunofluorescence assays for canine visceral leishmaniasis.
Revista do Instituto de Medicina Tropical de São Paulo
Several studies have recently shown the use of recombinant rabies virus as potential vector-viral... more Several studies have recently shown the use of recombinant rabies virus as potential vector-viral vaccine for HIV-1. The sequence homology between gp 120 and rabies virus glycoprotein has been reported. The McCoy cell line has therefore been used to show CD4+ or CD4+ like receptors. Samples of HIV-1 were isolated, when plasma of HIV-1 positive patients was inoculated in the McCoy cell line. The virus infection was then studied during successive virus passages. The proteins released in the extra cellular medium were checked for protein activity, by exposure to SDS Electrophoresis and blotting to nitro-cellulose filter, then reacting with sera of HIV positive and negative patients. Successive passages were performed, and showed viral replication, membrane permeabilization, the syncytium formation, and the cellular lysis (cytopathic effect). Flow cytometry analysis shows clear evidence that CD4+ receptors are present in this cell line, which enhances the likelihood of easy isolation and replication of HIV. The results observed allow the use of this cell line as a possible model for isolating HIV, as well as for carrying out studies of the dynamics of viral infection in several situations, including exposure to drugs in pharmacological studies, and possibly studies and analyses of the immune response in vaccine therapies.
Revista do Instituto de Medicina Tropical de São …, 1998
We evaluated the components of the Fuenzalida-Palacios antirabies vaccine, which is till used in ... more We evaluated the components of the Fuenzalida-Palacios antirabies vaccine, which is till used in most developing countries in human immunization for treatment and prophylaxis. This vaccine is prepared from newborn mouse brains at 1% concentration. Even though the vaccine is considered to have a low myelin content, it is not fully free of myelin or of other undesirable components that might trigger adverse effects after vaccination. The most severe effect is a post-vaccination neuroparalytic accident associated with Guillain-Barré syndrome. In the present study we demonstrate how the vaccines produced and distributed by different laboratories show different component patterns with different degrees of impurity and with varying protein concentrations, indicating that production processes can vary from one laboratory to another. These differences, which could be resolved using a better quality control process, may affect and impair immunization, with consequent risks and adverse effects after vaccination. We used crossed immunoelectrophoresis to evaluate and demonstrate the possibility of quality control in vaccine production, reducing the risk factors possibly involved in these immunizing products.
Revista de Saúde Pública, 2004
No population-based studies have been conducted to show the potential for the use of virological ... more No population-based studies have been conducted to show the potential for the use of virological diagnosis of the rabies virus. The objective of the present study was to estimate accuracy parameters for the isolation of the rabies virus in McCoy cells as an alternative method and to compare this with the use of murine neuroblastoma (N2A) cells, which is considered to be a reference method.