Cut-off values of immunological tests to identify patients at high risk of severe lupus nephritis

Cut-off values for anti-dsDNA, anti-nucleosome and anti-C1q antibodies tests and for complement-mediated hemolytic activity (CH50) were explored to identify patients with high risk of developing severe lupus nephritis (LN). Forty-one patients with confirmed systemic lupus erythematosus (SLE) were identified; their levels for the three antibodies and complement had been measured on a same serum sample. These patients were classified based on the presence of renal involvem ent; sixteen had active proliferative LN. With the cut-off values accepted in the laboratory for SLE diagnosis (anti-dsDNA > 100 UI/ml, anti-nucleosome > 50 U/ ml or CH50 < 190 UCH50%) no significant differences were found between patients with and without LN. Anti-C1q > 40 U/ml showed a statistically significant association with LN and had 80% of specificity. Cut-off values for LN identified by Receiver Operating Characteristic curves (ROC) were higher for anti-dsDNA (> 455 IU/ml) and anti-nucleosome (>107 U/ml), lower for CH50 (< 150 UCH50%) and, for anti-C1q (> 41 U/ml) coincided with the cut-off values accepted for SLE. Anti-C1q > 134 U/ml had a 92% of specificity, 56% of sensibility and was associated with a fifteen-fold increased risk of LN. The simultaneous presence of anti-nucleosome > 107 U/ml and anti-C1q > 134 U/ml was associated with a 27-fold higher probability for LN. According to these results, the cut-off values used to detect SLE activity could be inadequate to identify patients at high risk of severe LN.

Se exploraron valores de corte para los ensayos de anti-ADNdc, anti-nucleosoma, anti-C1q y complemento hemolítico total (CH50) capaces de identificar los casos con mayor riesgo de nefritis lúpica (NL) grave. Se seleccionaron 41 pacientes ≥ 16 años con lupus eritematoso sistémico (LES) confirmado que tenían titulados los niveles de los tres anticuerpos y CH50, en una misma muestra de suero. Fueron clasificados según presencia de compromiso renal; 16 presentaron formas proliferativas de NL activa. Con los valores de corte aceptados por el laboratorio para el diagnóstico de LES (anti-ADNdc > 100 UI/ml, anti-nucleosoma > 50 U/ml o un CH50 < 190 UCH50%) no se encontraron diferencias significativas entre casos con y sin NL. Un anti-C1q > 40 U/ml tuvo una especificidad del 80% y mostró una asociación estadísticamente significativa con NL. Al aplicar curvas Receiver Operating Characteristic (ROC) para NL, se identificaron valores de corte más altos para anti-ADNdc (> 455 IU/ml) y anti-nucleosoma (> 107 U/ml), más bajo para CH50 (< 150 UCH50%) y para el anti-C1q (> 41 U/ml) coincidió con el aceptado para diagnóstico de LES. Un anti-C1q > 134 U/ml presentó una sensibilidad del 56%, una especificidad del 92% y se asoció con quince veces más riesgo de NL. La presencia simultánea de anti-C1q > 134 U/ml y anti-nucleosoma > 107 U/ml se asoció 27 veces más riesgo de NL. De acuerdo a estos resultados los valores de corte empleados para actividad en pacientes con LES podrían resultar inadecuados para identificar pacientes con mayor riesgo de NL grave.

Synthesis and in silico study of 2-furyl(4-{4-[(substituted)sulfonyl]benzyl}-1-piperazinyl)methanone derivatives as suitable therapeutic agents

Abstract In the study presented here, a new series of 2-furyl(4-{4-[(substituted)sulfonyl]benzyl}-1-piperazinyl)methanone derivatives was targeted. The synthesis was initiated by the treatment of different secondary amines (1a-h) with 4-bromomethylbenzenesulfonyl chloride (2) to obtain various 1-{[4-(bromomethyl)phenyl]sulfonyl}amines (3a-h). 2-Furyl(1-piperazinyl)methanone (2-furoyl-1-piperazine; 4) was then dissolved in acetonitrile, with the addition of K2CO3, and the mixture was refluxed for activation. This activated molecule was further treated with equi-molar amounts of 3a-h to form targeted 2-furyl(4-{4-[(substituted)sulfonyl]benzyl}-1-piperazinyl)methanone derivatives (5a-h) in the same reaction set up. The structure confirmation of all the synthesized compounds was carried out by EI-MS, IR and 1H-NMR spectral analysis. The compounds showed good enzyme inhibitory activity. Compound 5h showed excellent inhibitory effect against acetyl- and butyrylcholinesterase with respective IC50 values of 2.91±0.001 and 4.35±0.004 µM, compared to eserine, a reference standard with IC50 values of 0.04±0.0001 and 0.85±0.001 µM, respectively, against these enzymes. All synthesized molecules were active against almost all Gram-positive and Gram-negative bacterial strains tested. The cytotoxicity of the molecules was also checked to determine their utility as possible therapeutic agents.

Convergent synthesis of new N -substituted 2-{[5-(1H -indol-3-ylmethyl)-1, 3, 4-oxadiazol-2-yl]sulfanyl}acetamides as suitable therapeutic agents

abstract A series of N-substituted 2-{[5-(1H-indol-3-ylmethyl)-1,3,4-oxadiazol-2-yl]sulfanyl}acetamides (8a-w) was synthesized in three steps. The first step involved the sequential conversion of 2-(1H-indol-3-yl)acetic acid (1) to ester (2) followed by hydrazide (3) formation and finally cyclization in the presence of CS2 and alcoholic KOH yielded 5-(1H-indole-3-yl-methyl)-1,3,4-oxadiazole-2-thiol (4). In the second step, aryl/aralkyl amines (5a-w) were reacted with 2-bromoacetyl bromide (6) in basic medium to yield 2-bromo-N-substituted acetamides (7a-w). In the third step, these electrophiles (7a-w) were reacted with 4 to afford the target compounds (8a-w). Structural elucidation of all the synthesized derivatives was done by 1H-NMR, IR and EI-MS spectral techniques. Moreover, they were screened for antibacterial and hemolytic activity. Enzyme inhibition activity was well supported by molecular docking results, for example, compound 8q exhibited better inhibitory potential against α-glucosidase, while 8g and 8b exhibited comparatively better inhibition against butyrylcholinesterase and lipoxygenase, respectively. Similarly, compounds 8b and 8c showed very good antibacterial activity against Salmonella typhi, which was very close to that of ciprofloxacin, a standard antibiotic used in this study. 8c and 8l also showed very good antibacterial activity against Staphylococcus aureus as well. Almost all compounds showed very slight hemolytic activity, where 8p exhibited the least. Therefore, the molecules synthesized may have utility as suitable therapeutic agents.

resumo Uma série de acetamidas 2-{[5-(1H-indol-3-ilmetil)-1,3,4-oxadiazol-2-il]sulfanila} N-substituídas (8a-w) foi sintetizada em três fases. A primeira etapa envolveu a conversão sequencial de ácido 2-(1H-indol-3-il)acético (1) a éster (2), seguido por hidrazida (3) e, finalmente, a e ciclização na presença de CS2 e KOH alcoólico produziu 5-(1H-indol-3-il- metil)-1,3,4-oxadiazole-2-tiol (4). Na segunda etapa, aminas arílicas/aralquílicas(5a-w) reagiram com brometo de 2-bromoacetila (6​​), em meio básico, para se obter acetamidas 2-bromo-N-substituídas (7a-w). Na terceira etapa, estes eletrófilos (7a- w) reagiram com 4, para se obter os compostos alvo (8a-w). A elucidação estrutural de todos os derivados sintetizados foi realizada por 1H-NMR, IR e técnicas de espectrometria de EI-MS. Além disso, eles foram submetidos a triagem de atividade antibacteriana e hemolítica. Análise da inibição enzimática foi bem apoiada pelos resultados de docking molecular. Por exemplo, o composto 8q exibiu melhor potencial inibitório contra α-glicosidase, e os compostos 8g e 8b exibiram, comparativamente, melhor inibição contra butirilcolinesterase (BChE) elipoxigenase (LOX), respectivamente. Do mesmo modo os compostos 8b e 8c mostraram excelente potencial antibacteriano contra SalmonellaTyphi, semelhante ao do ciprofloxacino, antibiótico padrão usado neste estudo. Os compostos 8c e 8l também mostraram excelente potencial antibacteriano contra Staphylococcus aureus . Quase todos os compostos mostraram pequena atividade hemolítica, sendo que o composto 8p apresentou menor atividade. Assim, as moléculas sintetizadas podem ter a sua utilidade como agentes terapêuticos adequados.

Characteristics of hemolytic activity induced by skin secretions of the frog Kaloula pulchra hainana

Background: Previous works had shown that scorpion venom induced neurotransmitter elevation and an inflammatory response associated with various anatomo-pathological modifications. The most dangerous scorpions species in Algeria responsible for these effects are Androctonus australis hector (Aah) and Androctonus amoreuxi (Aam). Results: Comparison of the physiopathological effects induced by the two venoms showed differences in the kinetic of cytokine release and in lung injury. The lung edema was only observed in response to Aah venom and it was correlated with cell infiltration. In order to better understand the involved mechanism in inflammatory response, we used two antagonists, atropine (non-selective muscarinic antagonist) and propranolol (ß adrenergic antagonist), which lead to a decrease of cell infiltration but has no effect on edema forming. Conclusion: These results suggest another pathway in the development of lung injury following envenomation with Aam or Aah venom.(AU)

Chemical constituents of Rabdosia japonica var. glaucocalyx and their anti-complementary activity / 中国中药杂志

To study the chemical constituents of Rabdosia japonica var. glaucocalyx and their anti-complementary activity on the basis of preliminary studies. Target isolation guided by anti-complementary activity test, compounds in the chloroform and n-butanol fractions were isolated and purified by silica gel and Sephadex LH-20 column chromatographies, and preparative HPLC. The structures were identified by various spectroscopic data including ESI-MS, 1H-NMR and 13C-NMR data. The compounds were evaluated for anti-complementary activity in vitro. Eleven compounds were isolated from the chloroform and n-butanol soluble fractions and identified as stigmasterol (1), stigmas-9 (11) -en-3-ol (2), glaucocalyxin D (3), kamebakaurin (4), maslinic acid (5), corosolic acid (6), minheryins I (7), diosmetin (8), caffeic acid ethylene ester (9), caffeic acid (10) and vitexin (11). Isoquercetrin, rutin, quercetin, 3-methylquercetin, luteolin, 7-methylluteolin, and apigenin which were isolated from the preliminary studies together with compounds 9 and 10 showed inhibition of the complement system by the classical pathway. Compounds 2, 4, 6-9 and 11 were obtained from this plant for the first time. Caffeic acid (10) showed the strongest activity in vitro with a CH50 value of 0.041 g x L(-1).

Suero de conejo con actividad de complemento para tipificación del complejo mayor de histocompatibilidad humano / Rabbit serum with complement activity to typify human major histocompatibility complex

En el ensayo de microlinfocitotoxicidad, el suero de conejo como fuente de complemento desempeña un rol esencial en la clasificación del complejo mayor de histocompatibilidad humano, tanto para la tipificación antigénica como en el cross-match linfocitario de la pareja donante-receptor antes de realizar los trasplantes. En este trabajo, se obtuvieron 3 lotes experimentales de dicho hemoderivado con óptimos indicadores de rendimiento, actividad biológica y estabilidad. El exanguíneo de los animales se realizó por la técnica de yugulación. El suero fue separado por centrifugación en condiciones ambientales y de temperatura controladas. Posteriormente, el producto se sometió a filtración esterilizante y se tomaron muestras para los ensayos de calidad. El estudio mostró que el producto conserva la actividad biológica al menos durante 18 meses y que los valores de citotoxicidad se encuentran dentro de los límites de aceptación para su empleo en los ensayos de histocompatibilidad pretrasplante

Microlinfocitotoxicity assay is one of the serological methods used to classify human major histocompatibility complex. In this technique the rabbit´s serum as complement source, plays an essential role in antigens determination and in lymphocytes cross match assay as necessary stage before the selection of the best donor-receiver pair to realize the organ transplant. Three experimental lots of normal rabbit serum were developed with excellent indicators of efficiency, biological activity and stability. Rabbit's blood was obtained through jugulating technique and serum was separated by centrifugation in controlled environment and temperature´s conditions. The sterilization process was performed by filtration and the samples were taken for quality´s control assays. Stability studies showed that the product kept the biological activity at least during 18 months after it was processed and cytotoxicity's values were adequate for its employment in histocompatibility pre-transplant assays

Virulence potential of filamentous fungi isolated from poultry barns in Cascavel, Paraná, Brazil / Potencial de virulência de fungos filamentosos isolados de celeiros de aves em Cascavel, Paraná, Brasil

Opportunistic fungi are those that normally would not cause diseases in otherwise healthy people, but are able to cause problems under some circumstances, and for this they need to possess a certain virulence potential. The objective of this study was to identify samples of filamentous fungi isolated from poultry barns in Cascavel, Paraná, and also to evaluate their virulence potential by assessing proteinase production, hemolytic activity, urease production, and growth rate at 37 ºC. We have evaluated the following samples: Acremonium hyalinulum (1 sample), Aspergillus sp. (12), Beauveria bassiana (1), Curvularia brachyspora (1), Paecilomyces variotti (1), and Penicillium sp. (2). Out of the 18 samples analyzed, 44.4 percent showed proteolytic activity using albumin as the substrate versus 66.7 percent when using casein; 66.7 percent showed hemolytic activity, 83.3 percent were positive for urea, and 88.9 percent grew at a temperature of 37 ºC. The results demonstrated that the majority of the isolates expressed virulence factors. Therefore, these isolates have the potential to harm human hosts, such as those working at poultry barns, especially predisposed or susceptible individuals.

Fungos oportunistas são aqueles que normalmente não causariam doenças em pessoas saudáveis, mas eles são capazes de causar problemas sob certas circunstâncias e, para isso, eles necessitam possuir algum potencial de virulência. O objetivo deste trabalho foi identificar amostras de fungos filamentosos isolados de granjas de aves em Cascavel, Paraná, e também avaliar o seu potencial de virulência, verificando a produção de proteinase, atividade hemolítica, produção de urease e crescimento a 37 ºC. Foram avaliados Acremonium hyalinulum (01), Aspergillus sp (12), Beauveria bassiana (01), Curvularia brachyspora (01), Paecylomices variotti (01) e Penicillium sp (02). Das 18 amostras, 44,4 por cento apresentaram atividade proteolítica usando como substrato a albumina e 66,7 por cento com caseína; 66,7 por cento demonstraram atividade hemolítica, 83,3 por cento foram uréia positivas e 88,9 por cento cresceram em temperatura de 37 ºC. Os resultados demonstram que a maioria dos isolados expressaram fatores de virulência e, portanto, têm potencial para causar danos a hospedeiros humanos como os trabalhadores dos aviários, sobretudo em indivíduos predispostos ou suscetíveis.

Activation of the alternative complement pathway in canine normal serum by Paracoccidioides brasiliensis / Ativação da via alternativa do complemento em soro de cão normal por Paracoccidioides brasiliensis

The dimorphic fungus Paracoccidioides brasiliensis is the etiological agent of paracoccidioidomycosis, a human granulomatous disease. Recently the first case of natural disease in dogs was reported. The complement system is an important effector component of humoral immunity against infectious agents. Therefore, the aim of this study was to evaluate the activation of the dog alternative complement pathway by P. brasiliensis. Initially, the ability of erythrocytes of guinea pig, rabbit, sheep, chicken and swine to activate the dog alternative pathway was evaluated. The guinea pig erythrocytes showed the greatest capacity to activate dog alternative pathway. The alternative (AH50) hemolytic activity was evaluated in 27 serum samples from healthy dogs and the mean values were 87.2 AH50/ml. No significant differences were observed in relation to sex and age. The alternative pathway activation by P. brasiliensis was higher in serum samples from adult dogs when compared to puppies and aged dogs (p < 0.05). This is the first report of dog alternative complement pathway activation by P. brasiliensis and suggests that it may play a protective role in canine paracoccidioidomycosis.

O fungo dimórfico Paracoccidioides brasiliensis é o agente etiológico da paracoccidioidomicose, uma doença granulomatosa humana. Recentemente, foi relatado o primeiro caso da doença natural em cães. O sistema complemento é um importante componente efetor da imunidade humoral contra agentes infecciosos. Portanto, o objetivo deste trabalho foi avaliar a ativação da via alternativa do complemento canina pelo P. brasiliensis. Inicialmente, foi avaliada a capacidade de eritrócitos de cobaia, coelho, carneiro, galinha e suíno ativarem a via alternativa do complemento canino. Os eritrócitos de cobaia apresentaram maior capacidade de ativar a via alternative do complemento canino. A atividade hemolítica da via alternativa (AH50) foi avaliada em 27 amostras de soro de cães saldáveis e os valores médios observados foram de 87,2 AH50/ml. Não foi observada diferença significativa ao sexo e idade. A ativação da via alternativa pelo P. brasiliensis foi maior nas amostras de soro de cães adultos quando comparada aos cães filhotes e idosos (p < 0.05). Este é o primeiro relato da ativação da via alternative do complemento canino pelo fungo P. brasiliensis e sugere que pode ter um papel protetor na paracoccidioidomicose canina.

Protein-energy malnutrition alters the C3 complement factor in response to lipopolysaccharide in a murine model / A desnutrição proteico-energética altera a concentração do fator C3 do sistema complemento em resposta ao lipopolissacarídeo em um modelo murínico

Malnutrition modifies the hostfis resistance to infection, altering many physiological processes, including hematopoiesis and immunological functions. In this study, we evaluated the total complement system and its C3 component factor in animals subjected to a protein malnutrition model with lipopolysaccharide (LPS) stimulation. The cellularity of blood, bone marrow and spleen, as well as the production of C3 and CH50, were evaluated. Two-month old male Swiss mice were submitted to protein malnutrition with a low-protein diet containing 4% protein. A control group received a control diet containing 20% protein. When the experimental group had reached a loss of about 20% in their original body mass, they were intravenously inoculated with LPS. Cells in the blood, bone marrow and spleen were counted and the circulating levels of C3 and CH50 were evaluated in these animals. Malnourished animals presented anemia, leucopenia, and a severe reduction in bone marrow and spleen cellularity. The survival rate of the malnourished animals was lower, as well as the production of C3 and CH50, if compared to the control animals. These findings suggest that malnourished mice present a deficient response to LPS. The decrease in the complement synthesis may be partially responsible for the immunodeficiency observed. These data lead us to conclude that the nutritional status interferes in the immune response activation.

La desnutrición proteico-energética modifica la resistencia a la infección, alterando numerosos procesos fisiológicos, incluyendo la hematopoyesis y la función inmunológica. En este estudio medimos las concentraciones séricas del factor C3 y del Sistema Complemento Total (CH50) en ratos con desnutrición proteico-energética estimulados con lipopolisacárido (LPS). Fue evaluada la celularidad periférica, medular y esplénica. Ratos Swiss, machos, adultos, con dos meses de edad fueron sometidos a desnutrición proteica con una dieta de 4% de proteína .El grupo control consumía una dieta con 20% de proteínas. Cuando los animales desnutridos perdieron 20% de su peso inicial, fueron inoculados por vía endovenosa con LPS. Fueron determinadas las concentraciones de células sanguíneas, de la médula ósea, del bazo y los valores de C3 y CH50 circulantes en los animales estimulados. Los animales desnutridos presentaron anemia y leucopenia además de una reducción significativa de celularidad de la médula ósea y del bazo. La sobrevivencia de este grupo era menor y también eran más bajas las concentraciones del factor C3 del complemento y del complemento total, en relación a los animales del grupo control. Los resultados sugieren que animales desnutridos muestran una respuesta deficiente a LPS. La síntesis menor de complemento puede ser en parte responsable por la inmunodeficiencia observada. Estos resultados nos conducen a inferir que la desnutrición proteico-energética interfiere en la activación de la respuesta inmune.

A desnutrição proteico-energética modifica a resistência à infecção, modi? cando diversos processos fisiológicos, incluindo a hematopoiese e as funções imunológicas. Neste estudo, avaliamos as concentrações séricas do fator C3 e do Sistema Complemento total (CH50) em um modelo no qual camundongos submetidos à desnutrição proteico-energética são estimulados com lipopolissacarídeo (LPS), e avaliamos a celularidade periférica, medular e esplênica. Camundongos Swiss, machos, adultos, com dois meses de idade foram submetidos ao processo de desnutrição proteica com uma dieta contendo 4% de proteína em comparação aos animais controles com uma dieta contendo 20% de proteína. Quando os animais do grupo desnutrido alcançaram aproximadamente 20% de perda de peso, em relação ao inicial, foram inoculados endovenosamente com LPS. As células do sangue, da medula óssea e do baço foram quantificadas, bem como as concentrações circulantes de C3 e CH50 em animais estimulados com LPS. Os animais desnutridos apresentaram anemia e leucopenia, além de redução significativa da celularidade da medula óssea e do baço. Os animais desnutridos apresentaram menor taxa de sobrevivência, bem como das concentrações do fator C3 do complemento e do complemento total em relação aos animais controles. Estes resultados sugerem que animais desnutridos apresentam uma resposta deficiente aos LPS. A síntese menor do complemento pode ser em parte responsável pela imunodeficiência observada. Estes resultados conduzem-nos a inferir que a desnutrição proteico-energética interfere na ativação da resposta imune.

Serum complement 3 (C3) and complement 4 (C4) level in febrile neutropenia patients at Dr. Kariadi Hospital, Semarang and Dr. Moewardi Hospital, Surakarta.

AIM: to identify the serum complement 3 (C3) and complement 4 (C4) level in febrile neutropenia and non-febrile neutropenia patients. METHODS: this is a cross-sectional prospective study. Samples were collected from patients with febrile neutropenia as sample group and patients with neutropenia but without fever as control. Both groups were tested for serum complement 3 and complement 4 level, and the data were analyzed using student T-test. RESULTS: from 37 neutropenia patients, 23 were classified as febrile neutropenia group and 14 in non-febrile neutropenia as control group. Total mean neutrophil count was 653.22/ml serum in sample group and 594.36/ml serum in control group (p=0.575). Mean C3 level was 95.74 ug/dl in sample group and 130.00 ug/dl in control group, showing significant difference with p=0.031. The mean serum C4 level was 34.13 ug/ml in sample group and 34.00 ug/dl in control group, the difference is not significant with p=0.98. When sample C3 and C4 data were combined, the total level was 125.61 ug/ml, which was significantly lower than the total C3 and C4 in control group 184.07 ug/dl. (p=0.04). CONCLUSION: in febrile neutropenia there is significant decrease of serum C3 level compared to non-febrile neutropenia. Serum C4 level in febrile neutropenia group is lower than the non-febrile neutropenia group, but the difference is not significant.

Some influence factors in determination of serum enzyme in hepatitis patients / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study some factors (temperature, time, anticoagulate, reagent, apparatus) which influence the detection of serum enzyme.</p><p><b>METHODS</b>Serums obtained from same patient are determined in different conditions. Compare the statistical difference among each group.</p><p><b>RESULTS</b>The anticoagulant-heparinize Li influence the determination of ADA and CH50. If serums were sent in room temperature for 24 hours, the results will be different. Using regents from different factories will receive different results.</p><p><b>CONCLUSION</b>Different conditions and reagents will influence results. To obtain correct data, we must institute and perform standard regulation.</p>

Serum complement (C3, C4) levels in patients with acute myocardial infarction and angina pectoris.

Serum complement (C3, C4) levels in Libyan patients with acute myocardial infarction (AMI; 31 patients) and angina pectoris (AP; 11 patients) at the 1st day and 7th day of attack were estimated. A group of 26 healthy Libyans were taken as control subjects (CS). Serum C3 and C4 levels (mean +/- SD, mg/dl) were elevated at the 1st day in AMI as well as AP patients (C3 --> AMI1: 154.0 +/- 28.5, AP1: 152.0 +/- 45.0, CS: 132.0 +/- 8.0, ANOVA: p = 0.0072; C4 --> AMII1: 38 +/- 13, AP1: 37 +/- 17, CS: 29 +/- 6, ANOVA: p = 0.0160). No significant differences for the elevated C3 and C4 levels at the 1st day were observed between the two diseases groups (AMI1 vs AP1 --> C3: p = 0.879, C4: p = 0.818). At the 7th day, C3 and C4 levels were further elevated in AMI, while they remained at the similar elevated levels in AP (C3 --> AMI 7: 173.1 +/- 28.0, AP 7: 149.0 +/- 41.0, CS: 132.0 +/- 8.0, ANOVA: p = 0.0000; C4 --> AMI 7: 46.0 +/- 7.0, AP 7: 36.0 +/- 15.0, CS: 29.0 +/- 6.0, ANOVA: p = 0.0000). Again, no significance differences for the raised C3 and C4 levels at the 7th day was observed between AMI and AP patients (AMI 7 vs AP 7 --> C3: P = 0.059, C4: p = 0.06). The C3 elevation showed significant positive correlation in AMI group (r = 0.522, p = 0.003) while it was insignificant in AP patients (r = 0.037, p = 0.915). Regarding C4 levels, it was significantly correlated in AMI (r = 0.483, p = 0.006), and in AP, although it was positively correlated (r = 0.656, P = 0.028) the observed difference was not significant (t = 0.29, p = 0.778). In conclusion, serum C3 and C4 levels were more profoundly elevated in AMI compared to AP patients suggestive of an acute phase and inflammatory response.

Complement levels in Brazilian children during and after meningococcal meningitis

OBJETIVO: Avaliar a atividade funcional das vias clássica e alternativa do sistema complemento e os níveis de C3, C4 e fator B durante o primeiro episódio de infecção meningocócica e durante a convalescença. PACIENTES E MÉTODOS: Dez crianças brasileiras com idades entre 8 meses e 8 anos, admitidas de 1991 a 1993, com diagnóstico clínico-laboratorial de meningite meningocócica, foram estudadas durante infecção aguda (até 7 dias do diagnóstico) e no período de convalescença (entre 1 e 6 meses após). C3, C4 e fator B foram quantificados por nefelometria e a atividade lítica das vias clássica e alternativa foi avaliada por método cinético e expressa como tempo necessário para lisar 50% de uma suspensão de eritrócitos (T1/2, expresso em segundos). Baixos valores de T1/2 das vias clássica e alternativa se correlacionam com elevadas atividades de via clássica e via alternativa, respectivamente. RESULTADOS: Observaram-se diferenças significativas entre a atividade lítica da via alternativa durante a infecção e no período de convalescença (282 e 238 segundos, respectivamente, P= .01). Nenhuma diferença foi detectada nos outros parâmetros analisados. CONCLUSÕES: Na presença de meningite meningocócica a via alternativa é preferencialmente ativada, provavelmente devido à maior capacidade da endotoxina meningocócica para ativar esta via, in vivo.

The role of complement in the early phase of Leishmania (Leishmania) amazonensis infection in BALB/c mice

Complement-depleted and -non-depleted BALB/c mice were inoculated with Leishmania (Leishmania) amazonensis promastigotes into the hind footpad to study the role of the complement system in cutaneous leishmaniasis. Total serum complement activity was measured by hemolytic assay and C3 fragment deposit at the inoculation site was determined by direct immunofluorescence in the early period of infection, i.e., at 3, 24, 48 h and 7 days post-infection. The inflammatory reaction and the parasite burden were evaluated in the skin lesion at 7 and 30 days post-infection. Total serum complement activity decreased in the early phase of infection, from 3 to 24 h, in non-depleted mice compared to non-infected and non-depleted mice. C3 fragment deposit at the site of parasite inoculation was present throughout the period of infection in non-depleted mice. In contrast, no C3 fragment deposit was observed at the inoculation site in complement-depleted mice. Complement-depleted mice showed a significant decrease in the inflammatory response and a significant increase in the number of parasites (70.0 ± 5.3 vs 5.3 ± 1.5) at 7 days of infection (P < 0.05). A higher number of parasites were also present at 30 days of infection at the inoculation site of complement-depleted mice (78.5 ± 24.9 vs 6.3 ± 5.7). These experiments indicate that complement has an important role at the beginning of experimental cutaneous leishmaniasis caused by L. (L.) amazonensis by controlling the number of parasites in the lesion.

Niveles de complemento hemolítico total, C3 y C4 en suero y liquído ascítico de pacientes con enfermedad crónica del hígado / Levels of C3 and C4, total hemolytic complement in serum and ascitic fluid of patients with chronic liver disease

Se estudiaron los niveles de complemento hemolítico total y sus fracciones C3 y C4 en sangre y líquido ascítico de pacientes con enfermedad crónica del hígado. Se ha referido que los pacientes con peritonitis bacteriana espontánea (PBE) presentan un descenso en líquido ascítico de los parámetros antes mencionados. En el presente estudio se incluyeron 29 pacientes 7 de los cuales tenían PBE. Los niveles de complmento hemolítico total, C3 y C4 en sangre y en líquido ascítico no mostraron diferencias significativas en pacientes con y sin PBE. En cambio, cuando se agruparon según el origen de la hepatopatía crónica en alcohólica o no alcohólica, en los pacientes con hepatopatía crónica no alcohólica los níveles de complemento hemolítico total en suero, C3 en suero y líquido ascítico fueron menores que en aquellos con hepatopatía crónica alcohólica.

Alteraciones del sistema complemento en pacientes con nefropatía diabética / Alterations of the complement system in patients with diabetic nephropathy

Se determinó la actividad del sistema complemento en 26 pacientes adultos que se hallaban en la fase terminal de la nefropatía diabética. En estos pacientes se observó una disminución significativa (p<0,001) de la actividad hemolítica de la vía clásica, de los niveles séricos del subcomponente Clq y de los niveles séricos del componente C4. Por el contrario, la actividad hemolítica de la vía alternativa, la actividad hemolítica del factor B y los niveles séricos del componente C3, se mantuvieron dentro del rango normal. Estos resultados sugieren que la disminución prolongada de alguno de los componentes iniciales de la vía clásica del complemento, puede constituir un factor que predispone a estos pacientes para los procesos autoinmunes, o que puede contribuir al mantenimiento del estado de autoinmunidad.

Recurrent pneumococcal meningitis in homozygous C3 deficiency.

Congenital deficiencies of complement system proteins are rare. A 4-year-old girl was admitted for meningitis. She had had repeated attacks of pneumococcal meningitis and otitis media at the age of 3 years. Analysis of cerebrospinal fluid showed that this meningitis was due to pneumococcal infection. Complement 3 and CH50 values of the proband and her brother were low, while her parents were normal. The patient was given polyvalent pneumococcal and anti-haemophilus vaccines plus ceftriaxone. Recovery was complete after 15 days of antibiotic therapy. This is the first description of a case of recurrent meningitis with C3 and CH50 deficiency in a Turkish family.

Producción de un anticuerpo de conejo anti-glóbulos rojos de carnero para utilizarlo en la valoración de la Actividad Hemolítica del Complemento (CH50) / Production of an anti-ram erythrocite rabbit antibody for use in haemolytic activity assessment of CH 50 complement

La valoración de la Actividad Hemolítica del Complemento (CH50) constituye un medio diagnóstico en el que se mide la capacidad de la vía clásica del complemento para lisar eritrocitos de carnero óptimamente sensibilizados con un anticuerpo anti-eritrocitos de carnero (hemolisina), cuya actividad esta en función de la cantidad y calidad de cada una de los componentes proteicos del complemento. Con el propósito de obtener una hemolisina capaz de sensibilizar adecuadamente a los eritrocitos de carnero, se sometieron a conejos adultos a inoculaciones repetidas y crecientes de antígeno de Forssman presentes en la superficie celular de los hematíes de carnero con lo cual se lograron inmunizar a los conejos contra este antígeno y obtener un anticuerpo en conejo contra eritrocitos de carnero, el cual fue titulado obteniéndose una dilución de 1:200 como título óptimo para la sensibilización de los eritrocitos de carnero, y su posterior utilización en la Valoración de la Actividad Hemolítica del Complemento.

Comportamiento de variables inmunológicas en pacientes politraumatizados / Inmunological variables behaviour in multiple trauma patients

Se realizó el estudio descriptivo de algunas variables de la inmunidad celular y humoral que incluyen las rosetas espontáneas y activas, las inmunoglobulinas, el complemento hemolítico total y los inmunocomplejos circulantes, en 30 pacientes politraumatizados atendidos en la unidad de terapia intensiva del Instituto Superior de Medicina Militar "Dr. Luis Díaz Soto". Los pacientes fueron monitoreados mediante 3 extracciones de sangre que correspondieron a las 24 h, a las 72 h y a los 7 d después de haber sufrido el trauma. Se produjo una disminución de las rosetas, cambios de los valores de inmunoglobulinas y aumento de los inmunocomplejos circulantes; por lo que el seguimiento de estas variables sería importante en la evaluación de los politraumatizados

Determinación de la actividad anti-C3d en reactivos antiglobulina humana / Determination of the anticomplement activity in human antiglobulin reagent

La evaluación de la actividad de anti-C3d en reactivos de Coombs poliespecíficos, con los procedimientos clásicos presenta, según la experiencia de los autores, dificultades técnicas y falta de reproductibilidad. El objetivo de este trabajo fue describir una técnica para controlar la actividad de los reactivos antiglobulina humana (AGH) poliespecíficos y monoespecíficos anti-C3d, utilizando glóbulos rojos de carnero (GRc) sensibilizados con fragmentos de complemento (C'). La obtención de GRc sensibilizados con C3d, se evaluó con reactivos de calidad certificada por la firma comercial. Se obtuvieron reacciones de aglutinación francamente positivas con reactivos de Coombs poliespecíficos y con reactivos anti-C3d, -C3d monoespecíficos. La sensibilidad, especificidad y sencillez de esta técnica induce a proponerla dentro del protocolo de Control de Calidad de los antisueros AGH poliespecíficos y anti-C3d, -C3d, utilizados de rutina en los Servicios de Medicina Transfusional