There is high incidence of skin cells in the first and third drops of cerebrospinal fluid in spinal anesthesia.

BACKGROUND AND OBJECTIVES: Skin fragments during lumbar punctures may develop intraspinal epidermoid tumors. The aim of this study was to determine the incidence of epithelial cells that reflow along with the first and third drops of CSF of patients undergoing spinal anesthesia. METHODS: Samples of the first and third drops of cerebrospinal fluid were collected from 39 adult patients undergoing spinal anesthesia with a 25G Quincke needle. Four microscope slides were prepared: one for the first drop, one for third drop, one for the needle, and one with a drop of saline for control. A pathologist examined the slides randomly. RESULTS: Squamous epithelial cells were identified in 35 (89.7%) samples from the first drop, 34 (87.2%) from the third drop, and 24 (61.5%) from spinal needle. The third drop showed a mean number of cells larger than the first drop (p=0.046). Nucleated epithelial cells were found in a sample of the first drop (2.56%), in four samples of third drop (10.25%), and in one spinal needle (2.56%). Third drop showed a mean number of nucleated cells higher than first drop with no statistical difference (p=0.257). CONCLUSIONS: High percentage of epithelial cells was found in the first (89.7%) and third (87.2%) drops of CSF reflow and in used needles (61.5%). Skin cells were found even using small gauge disposable needles with well-adapted mandrel.

Ocorre alta incidência de células da pele na primeira e terceira gotas do líquido cefalospinal em raquianestesia / There is high incidence of skin cells in the first and third drops of cerebrospinal fluid in spinal anesthesia / Ocurre una alta incidencia de células de la piel en la primera y tercera gotas del líquido cefaloespinal en la raquianestesia

JUSTIFICATIVA E OBJETIVOS: Fragmentos de pele durante punções subaracnóideas podem desenvolver tumores epidermoides intraespinais. O objetivo deste estudo foi verificar a incidência de células epiteliais que refluem junto com a primeira e terceira gotas de líquor de pacientes submetido a raquianestesia. MÉTODO: Foram obtidas amostras da primeira e terceira gotas de líquor em 39 pacientes adultos submetidos à raquianestesia com agulha 25G Quincke, sendo confeccionadas quatro lâminas: da primeira gota, da terceira gota, da agulha e uma quarta lâmina controle com uma gota de soro fisiológico. As lâminas foram examinadas de forma randomizada pelo patologista. RESULTADOS: Foram identificadas células epiteliais escamosas em 35 (89,7%) das amostras da primeira gota, em 34 (87,2%) da terceira gota e em 24 (61,5%) das agulhas espinais. A terceira gota apresentou em média maior número de células que a primeira gota (p = 0,046). Células epiteliais nucleadas foram encontradas em uma (2,56%) das amostras da primeira gota, em quatro (10,25%) da terceira gota e em uma (2,56%) das agulhas espinais. A terceira gota apresentou em média maior número de células nucleadas que a primeira gota sem diferença estatística (p = 0,257). CONCLUSÕES: Encontramos uma alta porcentagem de células epiteliais que refluem na primeira (89,7%) e na terceira (87,2%) gotas do líquor e nas agulhas utilizadas (61,5%). Mesmo utilizando agulhas de pequeno calibre, descartáveis e com mandril bem adaptado, foram encontradas células da pele.

BACKGROUND AND OBJECTIVES: Skin fragments during lumbar punctures may develop intraspinal epidermoid tumors. The aim of this study was to determine the incidence of epithelial cells that reflow along with the first and third drops of CSF of patients undergoing spinal anesthesia. METHODS: Samples of the first and third drops of cerebrospinal fluid were collected from 39 adult patients undergoing spinal anesthesia with a 25G Quincke needle. Four microscope slides were prepared: one for the first drop, one for third drop, one for the needle, and one with a drop of saline for control. A pathologist examined the slides randomly. RESULTS: Squamous epithelial cells were identified in 35 (89.7%) samples from the first drop, 34 (87.2%) from the third drop, and 24 (61.5%) from spinal needle. The third drop showed a mean number of cells larger than the first drop (p = 0.046). Nucleated epithelial cells were found in a sample of the first drop (2.56%), in four samples of third drop (10.25%), and in one spinal needle (2.56%). Third drop showed a mean number of nucleated cells higher than first drop with no statistical difference (p = 0.257). CONCLUSIONS: High percentage of epithelial cells was found in the first (89.7%) and third (87.2%) drops of CSF reflow and in used needles (61.5%). Skin cells were found even using small gauge disposable needles with well-adapted mandrel,.

JUSTIFICATIVA E OBJETIVOS: Algunos fragmentos de piel durante las punciones subaracnoideas pueden desarrollar tumores epidermoides intraespinales. El objetivo de este estudio fue verificar la incidencia de células epiteliales que refluyen junto con la primera y tercera gotas de líquido cefalorraquídeo de los pacientes sometidos a la raquianestesia. MÉTODO: Se obtuvieron muestras de la primera y tercera gotas de líquido cefalorraquídeo en 39 pacientes adultos sometidos a la raquianestesia con una aguja 25G Quincke, siendo confeccionadas cuatro láminas: de la primera gota, de la tercera gota, de la aguja y una cuarta lámina control con una gota de suero fisiológico. Las láminas fueron examinadas de forma aleatoria por el patólogo. RESULTADOS: Se identificaron células epiteliales escamosas en 35 (89,7%) de las muestras de la primera gota, en 34 (87,2%) de la tercera gota y en 24 (61,5%) de las agujas espinales. La tercera gota tuvo como promedio un mayor número de células que la primera gota (p = 0,046). Las células epiteliales nucleadas fueron encontradas en una (2,56%) de las muestras de la primera gota, en cuatro (10,25%) de la tercera gota y en una (2,56%) de las agujas espinales. La tercera gota presentó como promedio, un mayor número de células nucleadas que la primera gota sin diferencias estadísticas (p = 0,257). CONCLUSIONES: Encontramos un alto porcentaje de células epiteliales que refluyen en la primera (89,7%) y en la tercera (87,2%) gotas del líquido cefalorraquídeo y en las agujas utilizadas (61,5%). Y aunque hayamos usado agujas de pequeño calibre, desechables y con un mandril bien adaptado, se encontraron células de la piel.

There is high incidence of skin cells in the first and third drops of cerebrospinal fluid in spinal anesthesia.

BACKGROUND AND OBJECTIVES: Skin fragments during lumbar punctures may develop intraspinal epidermoid tumors. The aim of this study was to determine the incidence of epithelial cells that reflow along with the fi rst and third drops of CSF of patients undergoing spinal anesthesia. METHODS: Samples of the fi rst and third drops of cerebrospinal fluid were collected from 39 adult patients undergoing spinal anesthesia with a 25G Quincke needle. Four microscope slides were prepared: one for the fi rst drop, one for third drop, one for the needle, and one with a drop of saline for control. A pathologist examined the slides randomly. RESULTS: Squamous epithelial cells were identified in 35 (89.7%) samples from the fi rst drop, 34 (87.2%) from the third drop, and 24 (61.5%) from spinal needle. The third drop showed a mean number of cells larger than the fi rst drop (p = 0.046). Nucleated epithelial cells were found in a sample of the fi rst drop (2.56%), in four samples of third drop (10.25%), and in one spinal needle (2.56%). Third drop showed a mean number of nucleated cells higher than fi rst drop with no statistical difference (p = 0.257). CONCLUSIONS: High percentage of epithelial cells was found in the fi rst (89.7%) and third (87.2%) drops of CSF reflow and in used needles (61.5%). Skin cells were found even using small gauge disposable needles with well-adapted mandrel.

III Consenso Brasileiro para Pesquisa de Autoanticorpos em Células HEp-2: perspectiva histórica, controle de qualidade e associações clínicas / Third Brazilian Consensus for Autoantibodies Screening in HEp-2 Cells: historical perspective, quality control and clinical associations

OBJETIVO: O III Consenso Brasileiro para Pesquisa de Autoanticorpos em Células HEp-2 (FAN) objetivou discutir estratégias para controlar a qualidade do ensaio, promover a atualização das associações clínicas dos diversos padrões e avaliar as dificuldades de implantação do II Consenso ocorrido no ano de 2002. MÉTODOS: Nos dias 13 e 14 de abril de 2007 participaram do encontro em Goiânia pesquisadores e especialistas de diversos centros universitários e laboratórios clínicos de diferentes regiões do Brasil, com o propósito de discutir e aprovar as recomendações que visam a melhores padronização, interpretação e utilização do ensaio pelos clínicos. Foram convidados como ouvintes representantes comerciais de diferentes empresas produtoras de insumos para realização do teste de FAN. RESULTADOS E CONCLUSÃO: Dada a heterogeneidade de microscópios e reagentes disponíveis no mercado, o III Consenso enfatizou a necessidade do controle de qualidade em ensaios de imunofluorescência indireta. Foram também feitas algumas adequações na terminologia utilizada para classificar os diferentes padrões. Finalmente, foi realizada uma atualização das associações clínicas com finalidade de facilitar cada vez mais o melhor uso do ensaio pelos clínicos.

OBJECTIVE: The Third Brazilian Consensus for Autoantibodies Screening in HEp-2 Cells (ANA) had as purpose the evaluation of difficulties in the accomplishment of the 2nd Consensus recommendations that took place in the year of 2002, the discussion of strategies for quality control of the assay and the discussion of an update of the clinical associations of the several immunofluorescent patterns. METHODS: Several ANA experts from university centers and private laboratories in different areas in Brazil joined the workshop in Goiânia on 2007 April 13 and 14 with the purpose of discussing and approving the recommendations for standardization, interpretation and use of the test by physicians. Commercial representatives of different ANA slide brands were also invited as listeners to the workshop. RESULTS AND CONCLUSION: The 3rd ANA Consensus emphasized the need for quality control in indirect immunofluorescent assays since there is a considerable heterogeneity of available microscopes and reagents. It also promoted adaptations in the previously approved terminology used to classify the different patterns and finally updated the clinical associations of the several patterns with the purpose of providing guidance for interpretation of the assay by clinical pathologists and assistant physicians.

3º Consenso Brasileiro para pesquisa de autoanticorpos em células HEp-2 (FAN): recomendações para padronização do ensaio de pesquisa de autoanticorpos em células HEp-2, controle de qualidade e associações clínicas / Third Brazilian Consensus for autoantibodies screening in HEp-2 cells (ANA): recommendations for standardization of autoantibodies screening trial in HEp-2 cells, quality control and clinical associations

OBJETIVO: O 3º Consenso Brasileiro para pesquisa de autoanticorpos em Células HEp-2 (FAN) teve como propósito avaliar as dificuldades de implantação do 2º Consenso ocorrido no ano de 2002, discutir estratégias para controlar a qualidade do ensaio e promover a atualização das associações clínicas dos diversos padrões. MÉTODOS: Participaram do encontro em Goiânia nos dias 13 e 14 de abril de 2008 pesquisadores e especialistas de diversos centros universitários e laboratórios clínicos de diferentes regiões do Brasil, com o propósito de discutir e aprovar as recomendações que visam à melhor padronização, interpretação e utilização do ensaio pelos clínicos. Representantes comerciais de diferentes empresas produtoras de insumos para realização do teste de FAN foram convidados como ouvintes. RESULTADOS E CONCLUSÕES: O 3º Consenso enfatizou a necessidade do controle de qualidade em imunofluorescência dada a heterogeneidade de microscópios e reagentes disponíveis no mercado, promoveu adequações na terminologia utilizada para classificar os diferentes padrões e, finalmente, atualizou as associações clínicas com finalidade de facilitar cada vez mais o melhor uso do ensaio pelos clínicos.

OBJECTIVE: The Third Brazilian Consensus for autoantibodies Screening in HEp-2 cells had as purpose the evaluation of difficulties in the accomplishment of the 2nd Consensus recommendations that took place in the year of 2002, the discussion of strategies for quality control of the assay and the promotion of an update of the clinical associations of the several immunofluorescent patterns. METHODS: Several ANA experts from university centers and private laboratories in different areas in Brazil joined the workshop in Goiânia on 2008 April 13 and 14 with the purpose of discussing and approving the recommendations for standardization, interpretation and use of the test by physicians. Commercial representatives of different ANA slide brands were also invited as listeners to the workshop. RESULTS AND CONCLUSIONS: The 3rd Consensus emphasized the need for quality control in indirect immunofluorescent since there is a considerable heterogeneity of available microscopes and reagents. It also promoted adaptations in the previously approved terminology used to classify the different patterns and finally updated the clinical associations of the several patterns with the purpose of providing guidance for interpretation of the assay by clinical pathologists and assistant physicians.

Specific treatment for Trypanosoma cruzi: lack of efficacy of allopurinol in the human chronic phase of Chagas disease.

Thirty-five individuals from endemic areas of Central Brazil (age range, 18-64 years; 19 women) in the chronic phase of Chagas disease, with positive serology and presence of circulating parasites detected by one or more recent positive xenodiagnosis, were selected for this study. Allopurinol (900 mg/d) or placebo was administered in a double-blind clinical trial for 60 days. After codes were broken, 23 had been allocated to the intervention group and 12 to the placebo group. Side effects were observed in 11 patients in the intervention group and in 1 in the placebo group. Seventeen patients in the intervention group and 10 in the placebo group completed the trial. Follow-up was performed by monthly xenodiagnosis and serologic tests every 3 months during the first year and at the end of the trial. Xenodiagnosis remained positive in all 17 of the treated group and in all 10 of the placebo group. Serologic tests were persistently positive in both groups after treatment. We concluded that, at the doses used, allopurinol was not effective to clear, in our region, Trypanosoma cruzi from peripheral blood of infected individuals.

Busca retrospectiva da transmissão maternal da infecção chagásica em pacientes na fase crônica / A retrospective search for maternal transmission of Chagas infection from patients in the chronic phase

Avaliamos a transmissão maternal do Trypanosoma cruzi em 278 filhos de 145 mães com a doença crônica causada por esse protozoário, nascidos após o estabelecimento do diagnóstico materno. Usamos, nas mães e nos filhos, provas sorológicas como base do objetivo do estudo. Verificamos que ocorreram apenas duas veiculações pela modalidade citada (2/278 = 0,7 por cento). Como houve amamentação, não foi possível diferenciar, nesses casos, a transmissão congênita da adquirida pelo leite, mas, de qualquer forma, processou-se transmissão maternal, que constitui uma das modalidades alternativas de infecção pelo Trypanosoma cruzi. A metodologia empregada é mais uma dentre as diversas propostas para aquilatar a prevalência desse tipo de propagação do Trypanosoma cruzi. Ao lado da constatação fundamental, registramos fatos relacionados com a gestação, parto, puerpério, abortamento, prematuridade, nati-neomortalidade e aleitamento materno, que podem representar subsídios para melhores interpretações sobre o assunto.

[A retrospective search for maternal transmission of Chagas infection from patients in the chronic phase]. / Busca retrospectiva da transmissão maternal da infecção chagásica em pacientes na fase crônica.

Maternal transmission of Trypanosoma cruzi from 278 children of 145 mothers, chronically infected with this protozoan, to their offspring was investigated. This study was based upon serological tests. In only two cases (2/278 = 0.70%), such mode of transmission was demonstrated to have occurred. However, as according to extant records both patients had also been breast-fed, and the contribution of this factor could not be ruled out. In any case, maternal transmission, an alternative mode of acquiring the infection with Trypanosoma cruzi, was demonstrated. The methodology used is a further contribution to the evaluation of the prevalence of this propagating mechanism of T. cruzi; in addition to those aimed at the main objective of the investigation, records were kept about pregnancy, parturition, puerperium, abortion, prematurity, perinatal deaths and breast-feeding, which might contribute to a better interpretation of the subject.

Impacto do controle químico extensivo de Triatoma infestans sobre a incidência de casos agudos e a prevalência de doença de Chagas. O exemplo de Montalvânia, Minas Gerais / The impact of the extensive chemical control of Triatoma infestans on the incidence of acute cases and the prevalence of human Chagas disease. The example of Montalvânia, Minas Gerais State

A partir da emergência de um surto de casos agudos de doença de Chagas em Montalvânia, norte de Minas Gerais, Brasil, e da implantação do controle triatomínico com inseticidas, verifica-se pronta e conseqüente desaparição dos casos e redução da incidência da infecção no Município. É mostrada a evolução da implantação e do controle do Triatoma infestans em Montalvânia e a evolução da sorologia praticada na população em geral, ao longo dos últimos trinta anos.

[The impact of the extensive chemical control of Triatoma infestans on the incidence of acute cases and the prevalence of human Chagas disease. The example of Montalvânia, Minas Gerais State]. / Impacto do controle químico extensivo de Triatoma infestans sobre a incidência de casos agudos e a prevalência de doença de Chagas. O exemplo de Montalvânia, Minas Gerais.

From the emergency of acute cases of Chagas Disease in Montalvânia, Minas Gerais, Brazil, with the implementation of triatomine chemical control, the disappearing of these cases and the immediate reduction of incidence of the infection in the municipality are verified. The evolution of the dispersion and control of Triatoma infestans in Montalvânia and the evolution of the serology in general population during the last thirty years are showed.

Tratamento da fase crônica da doença de Chagas com nifurtimox associado a corticóide / Treatment of chronic Chagas' disease with an association of nifurtimox and corticoid

Dez pacientes com doença de Chagas em fase crônica foram tratados durante 60 dias, com uma única exceçäo, por meio de nifurtimox (8 a 9mg/kg/dia) conjuntamente com betametasona (doses progressivamente decrescentes a partir de 9mg/dia). Houve intençäo de utilizar a atividade antiparasitária do nifurtimox paralelamente à capacidade antiinflamatória do corticóide que, pela possibilidade de acentuar a infecção, poderia facilitar a açäo do fármaco anti-Trypanosoma cruzi. Em apenas um dos casos ocorreu negatividade prolongada dos xenodiagnósticos efetuados como controles e, quanto aos demais, ficaram evidentes os insucessos terapêuticos, corroborados pela persistência da positividade das provas sorológicas. O estudo também mostrou a necessidade de adequado e duradouro seguimento pós-tratamento para a obtençäo de corretas conclusöes

Double-blind study to evaluate flow cytometry analysis of anti-live trypomastigote antibodies for monitoring treatment efficacy in cases of human Chagas' disease.

The validation of flow cytometry analysis of anti-live trypomastigote antibodies (FC-ALTA) to monitor cure after treatment of Chagas' disease was evaluated with serum samples from treated and nontreated chagasic patients. After optimization of the original technique, toward better sensitivity and applicability to field surveys, we design a double blind study of 94 coded samples classified into the following categories: patients not treated (NT) and patients treated but not cured (TNC), both presenting positive conventional serology and xenodiagnosis; patients treated and cured (TC), showing negative serology and xenodiagnosis; and patients treated under evaluation (TUE), who presented positive or oscillating conventional serology (CSA) but negative xenodiagnosis. Coded samples, diluted 1:256, were assayed by incubation with live cell culture trypomastigotes, which were subsequently stained with fluorescein isothiocyanate-conjugated anti-human immunoglobulin G, with prior fixation and analysis by flow cytometry. The results were expressed as the percentages of positive fluorescent parasites (PPFP) for each individual sample, establishing 20% PPFP as the cutoff between negative and positive results. Our data demonstrated that all NT and TNC presented positive results while all but one TC had a PPFP lower than 20%. Analysis of TUE demonstrated a wide degree of reactivity, with PPFP values that were negative (PPFP 50%). As TUE with negative PPFP presented negative xenodiagnosis and positive or oscillating CSA, they were classified as dissociated according to the criteria of Krettli and Brener (J. Immunol. 128:2009-2012, 1982) and could indeed be considered cured after chemotherapy. This study demonstrates and validates the use of FC-ALTA to easily identify anti-live trypomastigote membrane-bound antibodies, offering another approach for investigating and monitoring the efficacy of specific chemotherapy in cases of human Chagas' disease.

[Correlation among the positivity of the artificial xenodiagnosis and the amount of blood and triatomines used in the exam, in chronic chagasic patients]. / Correlação entre a positividade do xenodiagnóstico artificial e a quantidade de sangue e triatomíneos utilizados no exame, em pacientes chagásicos crônicos.

The aim of this study was to verify whether the amount of blood and number of triotomines used could improve the artificial xenodiagnosis performed in 200 chronic phase infected individuals. Ten or 40ml of peripheral blood was collected in heparinized (20.4 IU) vacuum tubes, and fed to 60 and 360 triatomines, respectively. Dipetalogaster maximus (1st instar, about 15 days after eclosion), as well as 3rd instar Triatoma infestans and Triatoma vitticeps and the 4th instar of Rhodnius neglectus were used. The faecal examinations were performed 30 and 60 days after xenodiagnosis procedure. The positivity with 10ml of blood was 19% and with 40ml, 44%, from which it was concluded that a correlation existed between the amount of blood and the number of triatomines used (p < 0.01). The positivity of the xenodiagnosis ranged from 9.7 to 100%, higher in young and adults up to 34 years old, but independent in relation to the sex of the chronic chagasic individuals studied.

Correlação entre a positividade do xenodiagnóstico artificial e a quantidade de sangue e triatomíneos utilizados no exame, em pacientes chagásicos cronicos / Correlation among the positivity of the artificial xenodiagnosis and the amount of blood and triatomines used in the exam, in chronic chagasic patients

The aim of this study was to verify whether the amount of blood and number of triotomines used could improve the artificial xenodiagnosis performed in 200 chronic phase infected individuals. Ten or 40ml of peripheral blood was collected in heparinized (20.4 IU) vacuum tubes, and fed to 60 and 360 triatomines, respectively. Dipetalogaster maximus (1st instar, about 15 days after eclosion), as well as 3rd instar Triatoma infestans and Triatoma vitticeps and the 4th instar of Rhodnius neglectus were used. The faecal examinations were performed 30 and 60 days after xenodiagnosis procedure. The positivity with 10ml of blood was 19 and with 40ml, 44, from which it was concluded that a correlation existed between the amount of blood and the number of triatomines used (p < 0.01). The positivity of the xenodiagnosis ranged from 9.7 to 100, higher in young and adults up to 34 years old, but independent in relation to the sex of the chronic chagasic individuals studied.

[Treatment of chronic Chagas' disease with an association of nifurtimox and corticoid]. / Tratamento da fase crônica da doença de Chagas com nifurtimox associado a corticóide.

Ten patients with chronic Chagas' disease were treated with nifurtimox (8-9 mg/kg/day) associated with betamethasone (9 mg/day initially and then gradually reduced) during 60 days, with one exception. It was intended to combine the respective anti-parasitic and anti-inflammatory actions of these drugs. The expected stimulating effect of betamethasone on the infection could possibly enhance the anti-Trypanosoma cruzi action of nifurtimox. Long term persistence of negative xenodiagnosis, used to control the results, was observed in only one of the cases. Regaridng the other patients, post-treatment positivity of xenodiagnosis and serological testes attested the failure of this therapy. As this study has demonstrated, adequate and long term follow-up of treated cases is necessary to ensure correct conclusions.

Tentativa de tratamento específico, por meio da ticlopidina, de pacientes com doença de Chagas em fase crônica / An attempt of scpecifc treatment by means of ticlopidine of patients with chronic phase of Chagas' disease

Em investigaçäo experimental, a ticlopidina mostrou ser ativa no sentido de diminuir a parasitemia e a mortalidade quando avaliada a infecçäo de camundongos pelo Trypanosoma cruzi. Por isso, este fármaco foi administrado a 12 pacientes com doença de Chagas, em fase crônica. Houve utilizaçäo de 150, 200 ou 250mg, durante 90 dias, conforme se tratasse de crianças, adolescentes ou adultos, respectivamente. Ficou documentado cabal insucesso sob os pontos de vista parasitológico e sorológico