Lipidomics Detection of Brain Cardiolipins in Plasma Is Associated With Outcome After Cardiac Arrest.

OBJECTIVES: Brain mitochondrial dysfunction limits neurologic recovery after cardiac arrest. Brain polyunsaturated cardiolipins, mitochondria-unique and functionally essential phospholipids, have unprecedented diversification. Since brain cardiolipins are not present in plasma normally, we hypothesized their appearance would correlate with brain injury severity early after cardiac arrest and return of spontaneous circulation. DESIGN: Observational case-control study. SETTING: Two medical centers within one city. PARTICIPANTS (SUBJECTS): We enrolled 41 adult cardiac arrest patients in whom blood could be obtained within 6 hours of resuscitation. Two subjects were excluded following outlier analysis. Ten healthy subjects were controls. Sprague-Dawley rats were used in asphyxial cardiac arrest studies. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: We developed a high-resolution liquid chromatography/mass spectrometry method and determined cardiolipins speciation in human brain, heart, and plasma within 6 hours of (return of spontaneous circulation) from 39 patients with cardiac arrest, 5 with myocardial infarction, and 10 healthy controls. Cerebral score was derived from brain-specific cardiolipins identified in plasma of patients with varying neurologic injury and outcome. Using a rat model of cardiac arrest, cardiolipins were quantified in plasma, brain, and heart. Human brain exhibited a highly diverse cardiolipinome compared with heart that allowed the identification of brain-specific cardiolipins. Nine of 26 brain-specific cardiolipins were detected in plasma and correlated with brain injury. The cerebral score correlated with early neurologic injury and predicted discharge neurologic/functional outcome. Cardiolipin (70:5) emerged as a potential point-of-care marker predicting injury severity and outcome. In rat cardiac arrest, a significant reduction in hippocampal cardiolipins corresponded to their release from the brain into systemic circulation. Cerebral score was significantly increased in 10 minutes versus 5 minutes no-flow cardiac arrest and naïve controls. CONCLUSIONS: Brain-specific cardiolipins accumulate in plasma early after return of spontaneous circulation and proportional to neurologic injury representing a promising novel biomarker.

Molecular detection of Treponema pallidum sp. pallidum in blood samples of VDRL-seroreactive women with lethal pregnancy outcomes: a retrospective observational study in northern Brazil / Detecção molecular do Treponema pallidum sp. pallidum em amostras de sangue de mulheres sororeativas ao VDRL com resultado letal da gravidez: um estudo observacional retrospectivo no norte do Brasil

INTRODUCTION: Although control measures of maternal and congenital syphilis are available in Brazil, difficulties exist within the healthcare network in providing a laboratory diagnosis of the infection during the prenatal period. The objective of this study was to confirm the presence of Treponema pallidum by PCR in women with positive VDRL serology and lethal pregnancy outcomes, i.e., abortion, stillbirth and neonatal death. METHODS: A retrospective study was conducted on VDRLseroreactive women with lethal pregnancy outcomes admitted to the Fundação Santa Casa de Misericórdia do Pará (FSCM-PA) between January and July 2004. Serum samples and DNA from whole blood were obtained at the time of screening by the VDRL test. These samples were analyzed by IgG ELISA, IgM FTA-Abs and simple PCR (polA). RESULTS: During the study period, 0.7% (36/4,912) of women with lethal pregnancy outcomes presented a positive VDRL test. The polAgene was amplified in 72.7% (24/33) of these women, with 55.6% (20/36) and 94.4% (34/36) presenting IgM and IgG antibodies against T. pallidum, respectively. Comparison of these results showed a significant difference, with agreement between the PCR and IgM FTA-Abs results, suggesting that maternal syphilis was an active infection. No basic cause of death of the conceptus was reported in 97.2% (35/36) of cases. Among women who were submitted to the VDRL test during the prenatal period, only four of the nine seroreactive patients underwent treatment. CONCLUSIONS: The high frequency of syphilis in the group studied indicates the fragility of the service of infection diagnosis, treatment and monitoring, compromising epidemiological control.

INTRODUÇÃO: Apesar das medidas de controle da sífilis materna e congênita estarem disponíveis no Brasil, existem dificuldades da rede em prover o diagnóstico laboratorial da infecção durante o pré-natal. O objetivo deste estudo foi confirmar a presença do Treponema pallidum pela PCR em mulheres com sorologia positiva ao VDRL e com resultado letal da gravidez, isto é, aborto, natimorto e neomorto. MÉTODOS: Estudo retrospectivo realizado em mulheres VDRL-sororeativas com resultado negativo da gravidez, admitidas na Fundação Santa Casa de Misericórdia do Pará FSCM-PA entre janeiro e julho de 2004. As amostras de soro e DNA de sangue total foram obtidas no mesmo período da triagem pelo VDRL. Estas amostras foram analisadas pelo ELISA IgG, FTA-Abs IgM e PCR simples (polA). RESULTADOS: Durante o período de estudo, 0,7% (36/4.912) das mulheres com resultado letal da gravidez apresentaram VDRL positivo. O genepolA foi amplificado em 72,7% (24/33) destas mulheres,com 55,6% (20/36) e 94,4% (34/36) apresentando anticorpos tipo IgG e IgM contra o T. pallidum, respectivamente. A comparação destes resultados mostrou uma diferença estatística significativa, sendo que os resultados da PCR versus FTA-Abs Ig Mmostraram-se concordantes, sugerindo que a sífilis materna era uma infecção ativa. A causa básica de morte dos conceptos não foi relatada em 97,2% (35/36) dos casos. Entre as mulheres que foram submetidas ao VDRL no pré-natal, somente quatro das nove soropositivas receberam tratamento. CONCLUSÕES: A elevada frequência de sífilis no grupo de estudo indica a fragilidade do serviço no diagnóstico, tratamento e monitoramento da infecção, comprometendo o controle epidemiológico.

Molecular detection of Treponema pallidum sp. pallidum in blood samples of VDRL-seroreactive women with lethal pregnancy outcomes: a retrospective observational study in northern Brazil.

INTRODUCTION: Although control measures of maternal and congenital syphilis are available in Brazil, difficulties exist within the healthcare network in providing a laboratory diagnosis of the infection during the prenatal period. The objective of this study was to confirm the presence of Treponema pallidum by PCR in women with positive VDRL serology and lethal pregnancy outcomes, i.e., abortion, stillbirth and neonatal death. METHODS: A retrospective study was conducted on VDRLseroreactive women with lethal pregnancy outcomes admitted to the Fundação Santa Casa de Misericórdia do Pará (FSCM-PA) between January and July 2004. Serum samples and DNA from whole blood were obtained at the time of screening by the VDRL test. These samples were analyzed by IgG ELISA, IgM FTA-Abs and simple PCR (polA). RESULTS: During the study period, 0.7% (36/4,912) of women with lethal pregnancy outcomes presented a positive VDRL test. The polAgene was amplified in 72.7% (24/33) of these women, with 55.6% (20/36) and 94.4% (34/36) presenting IgM and IgG antibodies against T. pallidum, respectively. Comparison of these results showed a significant difference, with agreement between the PCR and IgM FTA-Abs results, suggesting that maternal syphilis was an active infection. No basic cause of death of the conceptus was reported in 97.2% (35/36) of cases. Among women who were submitted to the VDRL test during the prenatal period, only four of the nine seroreactive patients underwent treatment. CONCLUSIONS: The high frequency of syphilis in the group studied indicates the fragility of the service of infection diagnosis, treatment and monitoring, compromising epidemiological control.

Neurosyphilis in the age of AIDS: clinical and laboratory features.

Neurosyphilis remains to be a challenging diagnostic possibility worldwide. The aim of our study was to identify and report the clinical and laboratory profile of neurosyphilis, comparing features of HIV-infected and HIV-negative patients. A retrospective investigation of all cases of neurosyphilis, defined as positive VDRL test on cerebrospinal fluid, diagnosed at Hospital das Clínicas, Ribeirão Preto School of Medicine between January 1988 and December 2005, was carried out. We identified 35 patients with a mean age of 42.1 years, 28.6% of them HIV infected and 74.3% of them were male. HIV-infected patients were younger (34.6 years), presented with a higher frequency of the early forms of neurosyphilis, higher titers of serum VDRL and higher mean proteinorrachia at the suboccipital level. Neurosyphilis is still characterized by clinical polymorphism and there are significant differences in its epidemiological, clinical and laboratory profile when HIV-infected patients are compared with HIV-negative patients.

[Syphilis detection using ELISA and VDRL tests on blood donors at the blood center of Guarapuava, State of Paraná]. / Detecção de sífilis por ensaios de ELISA e VDRL em doadores de sangue do Hemonúcleo de Guarapuava, Estado do Paraná

The aim of this study was to evaluate the syphilis seroprevalence among 5,752 blood donors who were attended at the blood center of Guarapuava, State of Paraná, in 2006. The seropositivity rates were 2.1% for enzyme Linked Immuno Sorbent Assay and 0.2% for Veneral Disease Research Laboratory, thus showing low prevalence of syphilis among the individuals who came to this blood bank.

Detecção de sífilis por ensaios de ELISA e VDRL em doadores de sangue do Hemonúcleo de Guarapuava, Estado do Paraná / Syphilis detection using ELISA and VDRL tests on blood donors at the blood center of Guarapuava, State of Paraná

O objetivo desse estudo foi avaliar a soroprevalência de sífilis em 5.752 doadores de sangue atendidos no Hemonúcleo de Guarapuava-PR, em 2006. As taxas de positividade foram de 2,1 por cento pelo teste de ensaio imunoenzimático e 0,2 por cento pelo Veneral Disease Research Laboratory, mostrando baixa prevalência de sífilis nos indivíduos que procuraram este banco de sangue.

The aim of this study was to evaluate the syphilis seroprevalence among 5,752 blood donors who were attended at the blood center of Guarapuava, State of Paraná, in 2006. The seropositivity rates were 2.1 percent for enzyme Linked Immuno Sorbent Assay and 0.2 percent for Veneral Disease Research Laboratory, thus showing low prevalence of syphilis among the individuals who came to this blood bank.

Secondary syphilis with ocular manifestations in older adults.

We describe 4 patients (age, >50 years) with secondary syphilis. All patients presented with ocular syphilis, and 2 presented with syphilis-negative rapid plasma reagin titers due to a prozone phenomenon. Three male patients reported sildenafil use. The presentation of these patients with ocular syphilis suggests the need for additional clinical studies involving older patients.

[Prevalence of HIV-syphilis coinfection in a university hospital in the city of Rio de Janeiro in 2005]. / Prevalência da co-infecção HIV-sífilis em um hospital universitário da cidade do Rio de Janeiro no ano de 2005.

This was a study on the prevalence of HIV-syphilis coinfection among 830 HIV/AIDS patients who were being followed up as outpatients at a hospital in Rio de Janeiro between January and May 2005. The participants underwent laboratory tests at the venereal disease research laboratory (VDRL) consisting of CD4+/CD8+ cell counts and viral load tests. They answered questions about their sociodemographic characteristics and past medical history of syphilis. The prevalence of syphilis was 2.7% (22 patients) and the ratio between men and women with coinfection was approximately 4:1. Homosexual men were the category most affected. We did not find any association between coinfection and age, schooling and the laboratory parameters tested. Out of the total number of patients with syphilis, 73% (16) reported previous treatment; of these, 14 (88%) were reinfected, while two (12%) underwent inappropriate treatment. The presence of HIV-syphilis coinfection among patients undergoing routine follow-up signals the need to advise them to adopt safe sex practices, during their outpatient visits.

Prevalência da co-infecção HIV-sífilis em um hospital universitário da cidade do Rio de Janeiro no ano de 2005 / Prevalence of HIV-syphilis coinfection in a university hospital in the city of Rio de Janeiro in 2005

Estudo de prevalência da co-infecção HIV-sífilis realizado com 830 pacientes em acompanhamento ambulatorial para HIV/aids entre janeiro e maio de 2005 no Hospital na cidade do Rio de Janeiro. Os participantes realizaram exames de VDRL (veneral disease research laboratory), contagens de células CD4+/CD8+ e de carga viral e responderam perguntas sobre características sócio-demográficas e história prévia de sífilis. A prevalência da sífilis foi de 2,7 por cento (22), a relação entre homens e mulheres co-infectados foi de 4:1, aproximadamente. Homossexuais masculinos foram os mais acometidos e não encontramos associação entre co-infecção e idade, escolaridade e parâmetros laboratoriais testados. Do total de casos com sífilis, 73 por cento (16) relataram tratamento prévio; destes, 14 (88 por cento) pacientes foram re-infectados, enquanto 2 (12 por cento) pacientes realizaram tratamento inapropriado. A presença de co-infecção HIV-sífilis em pacientes em acompanhamento rotineiro alerta-nos para necessidade de aconselhá-los a adotar práticas sexuais seguras durante os seus atendimentos ambulatoriais.

This was a study on the prevalence of HIV-syphilis coinfection among 830 HIV/AIDS patients who were being followed up as outpatients at a hospital in Rio de Janeiro between January and May 2005. The participants underwent laboratory tests at the venereal disease research laboratory (VDRL) consisting of CD4+/CD8+ cell counts and viral load tests. They answered questions about their sociodemographic characteristics and past medical history of syphilis. The prevalence of syphilis was 2.7 percent (22 patients) and the ratio between men and women with coinfection was approximately 4:1. Homosexual men were the category most affected. We did not find any association between coinfection and age, schooling and the laboratory parameters tested. Out of the total number of patients with syphilis, 73 percent (16) reported previous treatment; of these, 14 (88 percent) were reinfected, while two (12 percent) underwent inappropriate treatment. The presence of HIV-syphilis coinfection among patients undergoing routine follow-up signals the need to advise them to adopt safe sex practices, during their outpatient visits.

Evaluation of the Determine Rapid Syphilis TP assay using sera.

The Abbott Determine Rapid Syphilis TP assay is a treponemal test that can be used in resource-poor settings that lack laboratory facilities. However, this test has not been extensively evaluated. We measured its sensitivity and specificity by using stored serum specimens (n = 567) from all persons who tested Treponema pallidum hemagglutination assay (TPHA) positive (n = 250) or TPHA indeterminate (n = 17) in the year 2001 and the first 300 patients in 2001 who tested TPHA negative at the Evandro Chagas Research Institute in Rio de Janeiro, Brazil. This rapid assay was independently interpreted by three different observers. With TPHA results as the reference, sensitivity ranged between readers from 95.6 to 98.4% and specificity ranged from 97.3 to 95.7%. There was little interreader variability in the interpretation of results, with approximately 98% agreement for all reader combinations. Of samples from persons with human immunodeficiency virus (HIV) infection (n = 198), sensitivity was 96.9 to 99.2% and it was 94.4 to 96.3% among HIV-negative persons (n = 127). Specificity was 92.4 to 95.5% among HIV-positive persons and 97.2 to 100% among HIV-negative persons. We found this test to have high sensitivity and specificity and little interreader variability, indicating that it may be easily used in resource-poor settings without laboratory facilities. Further studies are needed using this test on whole blood and under the clinical conditions for which it is intended.

Anticardiolipin and anti-beta2-glycoprotein-I antibodies in hypertensive disorders of pregnancy.

BACKGROUND: This study was undertaken in order to determine the frequency of anticardiolipin (aCL) and anti-beta2-glycoprotein-I antibodies (anti-beta2 GP-I) in patients with hypertensive disorders of pregnancy. METHODS: We studied aCL (IgG and IgM) and IgG anti-beta 2 GP-I by ELISA in the serum and IgG aCL and anti-beta 2 GP-I in the urine of 125 women with hypertensive disorders of pregnancy (cases) (75 had preeclampsia, 25 had eclampsia, and 25 had chronic hypertension). One hundred and twenty seven normal women were used as controls. Antibody positivity was defined as above the 90(th) percentile of the controls. RESULTS: We found no differences in frequency of serum anti-beta 2 GP-I or serum and urinary aCL between cases and controls. In contrast, we found that frequency of IgG anti-beta 2 GP-I was higher in the urine of cases (26.1%) compared to controls (9.4%, p = 0.001). Strength of association was stronger for urinary anti-beta2-glycoprotein-I in women with preeclampsia (odds ratio [OR] = 4.3, 95% confidence limit [CI 95%] = 1.95-9.62, p <0.0001). Cases and the subgroup of preeclamptic patients also had higher titers of urinary IgG anti-beta 2 GP-I than control women (p <0.0001). CONCLUSIONS: Our work suggests that urine testing is necessary in order to ascertain whether antibodies to beta(2)GP-I are associated with preeclampsia.

[Preeclampsia-eclampsia: a change in utero-placental microcirculation. Immunologic, histologic, and biochemical features]. / Preeclampsia-eclampsia: una alteración en la microcirculación útero-placentaria. Aspectos inmunológicos, histológicos y bioquímicos.

BACKGROUND: To know the alterations in the microcirculation of the placenta, umbilical cord, as well as the immune and hemorrheologic disorders in preeclampsia-eclampsia. MATERIAL AND METHODS: Two groups were conformed, 30 patients each, all of them with pregnancy of more than 24-week gestation. Group A included patients with preeclampsia-eclampsia and group B (control group) included women with normal pregnancy. In all patients determinations of levels of platelets, fibrinogen, antinuclear antibodies, IgG and IgM anticardiolipin, VDRL were made; clotting times were determined, and histopathologic analyses (placenta, umbilical cord and uterus-placenta membranes) were performed. RESULTS: Platelet levels in the group A were normal in 40% and low in 60%. In group B they were normal in 83.3% and low in 16.7%. with p < 0.05. In group A fibrinogen was normal in 10% and high in 90%; in the group B it was normal in 62.1% and high 37.9%, with p < 0.05. In group A prothrombin time (PT) was normal in 40% and high in 60%; in group B it was normal in 76.7% and low in 23.3%, with p < 0.05. in group TPT was normal in 36.7% and high in 62.1%, with p > 0.05. VDRL was negative in the 100% of the women of group A and positive in the 3.3% of the controls with p > 0.05. The antinuclear antibodies were positive in 6.7% in group A, and in 23.3% in group B, p < 0.05. IgG anticardiolipin antibodies were negatives in the 100% in both groups and IgM antibodies were negative in 96.7% in the group B and 3.3% in group A, p > 0.05. Analysis of histopathologic and immune changes did not show statistic significance when comparing both groups. CONCLUSIONS: Statistical and clinical significance was observed only in the hemorrheologic changes (PT, TPT, fibrinogen and platelets) and in the newborn weight.

[The role of yaws in the serologic screening of treponematoses in blood donors in Martinique]. / La place du pian dans le dépistage sérologique des tréponématoses chez les donneurs de sang de la Martinique.

Using TPHA instead of VDRL for syphilis blood-screening since 1995 showed an important increase of positive blood donors in Martinique. Yaws, another treponema disease, has been present on the island until 1975-1980. Usual tests are unable to identify which type--venereal or non venereal--of treponema is involved. Our study, carried out from January 1995 to May 1999, compares actual serological and epidemiological characteristics of TPHA reactive donors to former studies. In our results, the frequency of reactive TPHA is about 1.04% in blood donations. Donors are carrying serological tracks of a past treponema disease with very low rate of antibodies, sometimes linked to yaws. Among donors aged 18 to 30, prevalence is low and is going to become similar to the rate observed in Continental France. This means that this problem will disappear in new donor generations. We suggest the possibility for them to continue blood donation, if their personal preliminary enquiry fits the admission criteria for blood giving.

[Pyoverdin from Pseudomonas fluorescens: binding to the globulin fraction of serum and its effect on serological reaction titers]. / Pioverdina de Pseudomonas fluorescens: unión a la fracción globulina del suero y efecto sobre los títulos de reacciones serólogicas.

Pseudomonas fluorescens grows and produces pigment in refrigerated human blood at 4 degrees C. Saline precipitation of plasma showed that globulin and albumin fractions retained pyoverdin at different concentrations. By dialysis it was possible to determine that the pigment attached or aggregated to the protein in total plasma as well as in the fraction obtained by saline precipitation. A greater binding was observed at the globulin fraction. Gamma-globulin immunological activity was reduced due to the interaction with pyoverdin, as much in agglutination reaction (VDRL) as in neutralization (Streptolysin O).

Síndrome antifosfolípido / Antiphospholipid syndrome

El síndrome antifosfolípido es un cuadro clínico caracterizado principalmente por una tríada constituida por trombosis venosa profunda a repetición, abortos frecuentes y trombocitopenia, asociado a la presencia de anticoagulante lúpico y anticuerpos anticardiolipinas circulantes. Puede o no asociarse a mesenquimopatías y su tratamiento está constituido por los anticoagulantes y los corticoides. En este artículo de puesta al día se ha tratado de entregar una visión global muy resumida de una patología que recién está entrando al campo de la clínica y que aún no adquiere toda la importancia que merece. En su conocimiento falta aún mucho camino que recorrer y numerosos aspectos por investigar. Sin embargo, es importante que el clínico tenga presente esta posibilidad diagnóstica tanto en sus formas secundarias como primarias