In Situ

Chagas' disease is caused by the protozoan parasite Trypanosoma cruzi. The immune system plays an important role in the reduction of parasite load, but may also contribute to the development of lesions observed during the chronic phase of the disease. We analyzed cytokines produced by inflammatory heart cells in 21 autopsy samples obtained from patients with Chagas' disease divided according to the presence or absence of heart failure (HF). Left ventricular sections were analyzed by immunohistochemistry using antibodies against human IL-4, IFN-γ, TGF-β, TNF-α, and NOS2. In situ mRNA expression was quantified by a Low Density Array. The number of IFN-γ-positive cells was significantly higher than IL-4 positive cells. TNF-α, TGF-β and NOS2 were detected in 65%, 62% and 94% of samples respectively. There was an association between TNF-α-producing cells and the presence of HF. Subjects with HF presented higher levels of STAT4 mRNA, whereas FoxP3 and STAT6 levels were similar in the two groups. A Th1 cytokine pattern predominated in the cardiac inflammatory cell infiltrate of Chagas’ disease patients associated with HF. High degree of fibrosis was associated with low NOS2 expression. These results support the idea that Th1 immune responses are involved in heart lesions of Chagas' disease patients

Fluorescent Markers: Proteins and Nanocrystals

This book chapter will comment on fluorescent reporter proteins and nanocrystals’ applicability as fluorescent markers. Fluorescent reporter proteins in the Drosophila model system offer a degree of specificity that allows monitoring cellular and biochemical phenomena in vivo, such as autophagy, mitophagy, and changes in the redox state of cells. Titanium dioxide (TiO2) nanocrystals (NCs) have several biological applications and emit in the ultraviolet, with doping of europium ions can be visualized in the red luminescence. Therefore, it is possible to monitor nanocrystals in biological systems using different emission channels. CdSe/CdS magic-sized quantum dots (MSQDs) show high luminescence stability in biological systems and can be bioconjugated with biological molecules. Therefore, this chapter will show exciting results of the group using fluorescent proteins and nanocrystals in biological systems

Production of cytokine and chemokines by human mononuclear cells and whole blood cells after infection with Trypanosoma cruzi

INTRODUCTION: The innate immune response is the first mechanism of protection against Trypanosoma cruzi, and the interaction of inflammatory cells with parasite molecules may activate this response and modulate the adaptive immune system. This study aimed to analyze the levels of cytokines and chemokines synthesized by the whole blood cells (WBC) and peripheral blood mononuclear cells (PBMC) of individuals seronegative for Chagas disease after interaction with live T. cruzi trypomastigotes. METHODS: IL-12, IL-10, TNF-α, TGF-β, CCL-5, CCL-2, CCL-3, and CXCL-9 were measured by ELISA. Nitrite was determined by the Griess method. RESULTS: IL-10 was produced at high levels by WBC compared with PBMC, even after incubation with live trypomastigotes. Production of TNF-α by both PBMC and WBC was significantly higher after stimulation with trypomastigotes. Only PBMC produced significantly higher levels of IL-12 after parasite stimulation. Stimulation of cultures with trypomastigotes induced an increase of CXCL-9 levels produced by WBC. Nitrite levels produced by PBMC increased after the addition of parasites to the culture. CONCLUSIONS: Surface molecules of T. cruzi may induce the production of cytokines and chemokines by cells of the innate immune system through the activation of specific receptors not evaluated in this experiment. The ability to induce IL-12 and TNF-α contributes to shift the adaptive response towards a Th1 profile

Cytokine and nitric oxide production in an adult patient with staphylococcal scalded skin syndrome. Cytokine and nitric oxide production in an adult patient with staphylococcal scalded skin syndrome

Abstract. Staphylococcal scalded-skin syndrome (SSSS) is an exfoliative dermatitis resultant from the infection with exfoliative-toxin-producing (ET) Staphylococcus aureus. This syndrome usually occurs in children, while adult cases are generally linked to renal-deficiency or immunossupresion. A case of a 74 years old woman presenting SSSS after hospital admission due to cardiovascular disorders is presented and discussed. Cytokines (TNF-a, IFN-g, IL-6, IL-13 and IL-10) and nitric oxide (NO) production in vitro by whole blood leukocytes (WBL) were investigated. Leukocytes stimulated by lipopolysaccharide or phytohemagglutinin produced increased IFN-g, TNF-a, IL-13 and NO levels after treatment. Based on these results, immunological aspects of the disease are discussed. Producción de citocinas y óxido nítrico en uno paciente adulto con síndrome de la piel escaldada estafilocócica. Invest Clin 2008; 49(4): 547 -552 Palabras clave: Síndrome de la piel escaldada, citocinas, óxido nítrico. Resumen. El síndrome de la piel escaldada estafilocócica (SSSS) es una dermatitis exfoliante resultante de la infección por Staphylococcus aureus productores de toxinas exfoliantes. Este síndrome ocurre generalmente en niños, mientras que su manifestación en adultos está generalmente relacionada con deficiencia renal o inmunosupresión. Se presenta y discute el caso de una mujer de 74 años de edad con SSSS luego de su admisión en el hospital por complicaciones cardiovasculares. Fue evaluada la producción in vitro de cito

Evaluation of the effectiveness of packed red blood cell irradiation by a linear accelerator

Irradiation of blood components with ionizing radiation generated by a specific device is recommended to prevent transfusion-associated graft-versus-host disease. However, a lin- ear accelerator can also be used in the absence of such a device, which is the case of the blood bank facility studied herein. In order to evaluate the quality of the irradiated packed red blood cells, this study aimed to determine whether the procedure currently employed in the facility is effective in inhibiting the proliferation of T lymphocytes without damaging blood components. The proliferation of T lymphocytes, plasma potassium levels, and the degree of hemolysis were evaluated and compared to blood bags that received no irradiation. Packed red blood cell bags were irradiated at a dose of 25 Gy in a linear accelerator. For this purpose, a container was designed to hold the bags and to ensure even distribution of irradiation as evaluated by computed tomography and dose-volume histogram. Irradiation was observed to inhibit the proliferation of lymphocytes. The percentage of hemolysis in irradiated bags was slightly higher than in non-irradiated bags (p-value >0.05), but it was always less than 0.4% of the red cell mass. Although potassium increased in both groups, it was more pronounced in irradiated red blood cells, especially after seven days of storage, with a linear increase over storage time. The findings showed that, at an appropriate dosage and under validated conditions, the irradiation of packed red blood cells in a linear accelerator is effective, inhibiting lymphocyte proliferation but without compromising the viability of the red cells

Provas de atividade inflamatória no pênfigo foliáceo endêmico

O pênfigo foliáceo endêmico (PFE) é uma afecção onde fenômenos auto-imunes são freqüentemente relacionados com sua patogênese. Abordaram-se neste estudo, alguns testes laboratoriais freqüentemente alterados em outras doenças auto-imunes. Foram estudados em 20 pacientes com PFE a presença ou alterações de fator antinúcleo (FAN), fator reumatóide (FR), proteína C-reativa (PCR), velocidade de hemossedimentação (VHS), eletroforese de proteínas e o número totalde leucócitos. Encontrou-se PCR positiva em mais de 65 % dos casos, leucocitose moderada na maioria dos pacientes, VHS aumentada e alterações discretas da análise de proteínas do sangue. O FAN e o FR apresentaram-se negativos em todos os casos. Embora estes exames sejam considerados inespecíficos, o seu estudo associado ao quadro clínico pode colaborar no acompanhamento do PFE