Intravitreal neurodegenerative and inflammatory mediators in proliferative diabetic retinopathy / Mediadores intravítreos da neurodegeneração e inflamação na retinopatia diabética proliferativa

ABSTRACT Purpose: To compare the intravitreal concentrations of cellular mediators involved in neurodegeneration, inflammation, and angiogenesis in patients with proliferative diabetic retinopathy and other vitreoretinal diseases. Methods: A multiplex bead immunoassay was used to measure vitreous levels of pigment epithelium-derived factor, serum amyloid P, C-reactive protein, complement C4, alpha-1 antitrypsin, vascular endothelial growth factor, platelet-derived growth factor-AA, platelet-derived growth factor-BB, interleukin-6, interleukin-8, interleukin-10, tumor necrosis factor alpha and beta in patients undergoing 23-gauge vitrectomy for proliferative diabetic retinopathy and other diagnoses (control group). Results: We evaluated 55 patients, of whom 24 had proliferative diabetic retinopathy and 31 had other diagnoses including vitreous hemorrhage, retinal detachment, macular hole, and epiretinal membrane. Patients with proliferative diabetic retinopathy demonstrated increased levels of serum amyloid P (85.49 vs. 31.38 ng/mL); C-reactive protein (59.89 vs. 41.75 ng/mL), vascular endothelial growth factor (2,330.11 vs. 554.25 pg/mL; p<0.001), platelet-derived growth factor A (127.32 vs. 39.11 pg/mL), platelet-derived growth factor B (29.37 vs. 7.12 pg/mL), interleukin-6 (69.37 vs. 33.58 pg/mL), interleukin-8 (175.25 vs. 59.71 pg/mL), and interleukin-10 (3.70 vs. 1.88 pg/mL); all p<0.004 when compared with the control group. Levels of pigment epithelium-derived factor (30.06 vs. 27.48 ng/mL; p=0.295), complement C4 (570.78 vs. 366.24 ng/mL; p=0.069), and alpha-1-antitrypsin (359.27 vs. 522.44 ng/mL; p=0.264) were not significantly different between the groups. Intravitreal levels of tumor necrosis factor-alpha and tumor necrosis factor-beta were undetectable. Serum Amyloid P, C-reactive protein, platelet-derived growth factor A, platelet-derived growth factor B, interleukin-6, and interleukin-8 were correlated positively with vascular endothelial growth factor. Conclusions: Cellular mediators involved in neurodegeneration and inflammation demonstrated increased levels in the vitreous humor of patients with proliferative diabetic retinopathy and may be part of the pathogenesis of diabetic retinopathy.

RESUMO Objetivo: Comparar as concentrações intravítreas de mediadores celulares envolvidos na neurodegeneração, inflamação e angiogênese em pacientes com retinopatia diabética proliferativa e outras doenças vítreo-retinianas. Métodos: Um ensaio imunomagnético foi utilizado para medir os níveis vítreos do fator derivado do epitélio pigmentar, amilóide P sérico, proteína-C-reativa, complemento C4, e alfa-1-antitripsina, fator de crescimento do endotélio vascular, fator de crescimento derivado das plaquetas AA, fator de crescimento derivado das plaquetas BB, interleucina-6, interleucina-8, interleucina-10, fator de necrose tumoral alfa e beta em pacientes submetidos à vitrectomia 23-gauge para retinopatia diabética proliferativa ou outros diagnósticos (grupo controle). Resultados: Foram avaliados 55 pacientes, dos quais 24 tinham retinopatia diabética proliferativa e 31 tinham outros diagnósticos, incluindo hemorragia vítrea, descolamento de retina, buraco macular e membrana epirretiniana. Pacientes com retinopatia diabética proliferativa demonstraram níveis aumentados de amilóide P sérico (85,49 vs 31,38 ng/mL), proteína-C-reativa (59,89 vs 41,75 ng/mL), fator de crescimento do endotélio vascular (2.330,11 vs 554,25 pg/mL, p<0.001), fator de crescimento derivado das plaquetas-A: (127,32 vs 39,11 pg/mL), fator de crescimento derivado das plaquetas-B (29,37 vs 7,12 pg/mL), interleucina-6 (69,37 vs 33,58 pg/mL), interleucina-8 (175,25 vs 59,71 pg/mL) e interleucina-10 (3,70 vs 1,88 pg/mL), todos com p<0,004 quando comparados ao grupo controle. Níveis de fator derivado do epitélio pigmentar (30,06 vs 27,48 ng/mL; p=0,295), complemento C4 (570,78 vs 366,24 ng/mL; p=0,069), alfa-1 antitripsina (359,27 vs 522,44 ng/mL; p=0,264) não foram significativamente diferente entre os grupos. Níveis intravítreos de fator de necrose tumoral alfa e fator de necrose tumoral beta foram indetectáveis. O amilóide P sérico, a proteína C-reativa, o fator de crescimento derivado das plaquetas A e B, a interleucina-6 e a interleucina-8 correlacionaram-se positivamente com o fator de crescimento do endotélio vascular. Conclusões: Os medidores celulares envolvidos na neurodegeneração e inflamação demonstraram níveis aumentados no humor vítreo de pacientes com retinopatia diabética proliferativa e podem ser parte da patogênese da retinopatia diabética.

B cells expressing IL-10 mRNA modulate memory T cells after DNA-Hsp65 immunization

In DNA vaccines, the gene of interest is cloned into a bacterial plasmid that is engineered to induce protein production for long periods in eukaryotic cells. Previous research has shown that the intramuscular immunization of BALB/c mice with a naked plasmid DNA fragment encoding the Mycobacterium leprae 65-kDa heat-shock protein (pcDNA3-Hsp65) induces protection against M. tuberculosis challenge. A key stage in the protective immune response after immunization is the generation of memory T cells. Previously, we have shown that B cells capture plasmid DNA-Hsp65 and thereby modulate the formation of CD8+ memory T cells after M. tuberculosis challenge in mice. Therefore, clarifying how B cells act as part of the protective immune response after DNA immunization is important for the development of more-effective vaccines. The aim of this study was to investigate the mechanisms by which B cells modulate memory T cells after DNA-Hsp65 immunization. C57BL/6 and BKO mice were injected three times, at 15-day intervals, with 100 µg naked pcDNA-Hsp65 per mouse. Thirty days after immunization, the percentages of effector memory T (TEM) cells (CD4+ and CD8+/CD44high/CD62Llow) and memory CD8+ T cells (CD8+/CD44high/CD62Llow/CD127+) were measured with flow cytometry. Interferon γ, interleukin 12 (IL-12), and IL-10 mRNAs were also quantified in whole spleen cells and purified B cells (CD43−) with real-time qPCR. Our data suggest that a B-cell subpopulation expressing IL-10 downregulated proinflammatory cytokine expression in the spleen, increasing the survival of CD4+ TEM cells and CD8+ TEM/CD127+ cells.

Use of minimally invasive gingival biopsies in the study of inflammatory mediators expression and their correlation with gingival fluid in patients with chronic periodontitis.

Background: There are no studies comparing the gingival crevicular fluid (GCF) cytokines expression with its corresponding values from the same tissue’s sites. Such comparison might be of great value since most of the cytokine function is related to cell and/or tissue receptors. Aims: Our aim was to use minimally invasive biopsies to evaluate the expression of interferon‑gamma, interleukin 1 (IL‑1) β, IL‑6, IL‑17A, IL‑17F, and their correlation with the expression in gingival fluid in patients with chronic periodontitis. Materials and Methods: The collection of gingival fluid comprised 22 samples from 11 patients (mean age 46.73 ± 10.16 standard deviation years) with chronic periodontitis. The collection of biopsies comprised 22 samples from the same patients. Gingival fluid and biopsy were taken from the same site in one shallow and one deep site per patient. Gingival fluid samples were collected with periopaper® and analyzed using Luminex®. Biopsies were taken with a 2 mm diameter punch and analyzed for the same mediators using immunohistochemistry. Results: The gingival fluid showed higher amounts for IL‑1‑β in deep sites. Immunohistochemical markers were observed in the analyzed cells groups, both in deep and shallow sites, without significant differences between them. In the comparative analysis between immunohistochemical markers and GCF, IL‑1‑β showed high concordance in shallow and deep sites. Conclusions: The use of a standardized punch of 2 mm diameter for periodontal tissue biopsies seems to be suitable for immunohistochemistry analysis and showed that the GCF may not express all the markers in the same proportion at the corresponding tissue.

A comparative study of pentoxifylline effects in adult and aged rats submitted to lung dysfunction by thermal injury

PURPOSE: To investigate the protective effects of pentoxifylline against lung injury observed after dorsal scald in aged animals. METHODS: Adult (eight months old) and aged (20 months old) rats were subjected to thermal injury or sham procedure. The six hours post-trauma animals received pentoxifylline and after 24 hours were euthanatized and lung tissue samples collectedted. The bronchoalveolar lavage fluid was evaluated for total protein content and tumor necrosis factor-alpha cytokine. Malondialdehyde and myeloperoxidase activety in the lung homogenate were measured and a histological lung examination was undertaken. RESULTS: Burn injury induced oxidative stress in lung homogenate was higher in elderly-burned rats compared to adult-burned rats (p<0.001). Total protein and cytokine in bronchoalveolar lavage increased in the elderly-burned group when compared to the adult-burned group (p<0.001). All parameters decreased in bolth groups treated with pentoxifylline (p<0.05). CONCLUSIONS: The injury was augmented in elderly rats when compared to adult rats. Damage was reduced with the use of pentoxifylline, however further studies are needed to evaluate the dose-response of the drug.

Marcadores periféricos e a fisiopatologia do transtorno bipolar / Peripheral markers and the pathophysiology of bipolar disorder

INTRODUÇÃO: O entendimento da fisiopatologia do transtorno bipolar vem tendo avanços consistentes nos últimos anos. Um enfoque na relação entre carga alostática e alterações sistêmicas vem tomando corpo, com o objetivo de se entender a frequente progressão da doença. Proeminentes entre os mediadores periféricos têm sido as moléculas que poderiam ser amplamente agrupadas em neurotrofinas, marcadores de estresse oxidativo e marcadores inflamatórios. OBJETIVO: Descrever achados recentes em relação à fisiopatologia sistêmica do transtorno bipolar, com enfoque especial em estudos brasileiros, tentando articular uma visão coerente do conhecimento atual do campo. MÉTODO: Revisão narrativa da literatura relacionada a neurotrofinas, estresse oxidativo e marcadores inflamatórios no transtorno bipolar. RESULTADOS: Diversas fontes de evidência, provenientes tanto de estudos pré-clínicos quanto clínicos, revelam consistentemente alterações sistêmicas no transtorno bipolar. Os achados são especialmente robustos em pacientes com múltiplos episódios. Nesses, alterações relacionadas a episódios de mania e depressão são notáveis em neurotrofinas e dano oxidativo a lipídeos. Um número menor de estudos mostra alterações no sistema imune, em particular estados pró-inflamatórios. CONCLUSÃO: Alterações sistêmicas que correlacionam o transtorno bipolar a comorbidade clínica, disfunção cognitiva, incapacidade e mortalidade precoce começam a ser traçadas. Estudos envolvendo desenhos longitudinais, amostras populacionais e ensaios clínicos envolvendo marcadores periféricos devem ser incorporados no futuro próximo e reforçar a validade de uma noção de envolvimento multissistêmico no transtorno bipolar.

INTRODUCTION: The understanding of the pathophysiology of bipolar disorder has steadily advanced in the past few years. Thereby, a focus on allostatic load and systemic changes has appeared, with the aim to understand illness progression. Amongst the peripheral markers, molecules that can be widely classified into neurotrophins, oxidadive stress markers, and inflammation markers have been elevated. OBJECTIVE: To describe recent findings regarding the systemic pathophysiology of bipolar disorder, with a special focus on Brazilian studies and to create a coherent view of the current knowledge in the field. METHOD: Narrative review of the literature regarding neurotrophins, oxidative stress, and inflammatory markers in bipolar disorder. RESULTS: A diverse body of evidence, based on both pre-clinical and clinical studies, reveals consistent systemic changes in bipolar disorder. The findings are particularly robust in patients after multiple episodes. Thereby, remarkable changes related to manic and depressive episodes were found in neurotrophins and oxidative damage to lipids. Regarding to immune system alterations, in particular pro-inflammatory states, the literature is less consistent. DISCUSSION: Systemic changes that link bipolar disorder to clinical comorbidity, cognitive dysfunction, disability and early mortality are becoming evident. In the near future, longitudinal studies with population-based samples and clinical trials incorporating biomarkers are needed to shed light upon the notion of a multisystem involvement in bipolar disorder.

Aterosclerosis en sujetos con periodontitis / Association between atherosclerosis and periodontitis

Background: Chronic infl ammation and infections are involved in the development and progression of atherosclerotic vascular disease. Aim: To evaluate the association between periodontitis and early atherosclerosis. Material and Methods: Fifty-three subjects who received periodontal treatment and regular maintenance for at least 10 years, and 55 subjects with periodontitis but without a history of periodontal treatment were studied. Carotid artery intima-media wall thickness (CIMT) was measured with high-resolution B-mode ultrasonography. A blood sample was obtained to measure high sensitivity C-reactive protein, fibrinogen, lipoprotein cholesterol, leukocyte count and erythrocyte sedimentation rate. Covariates included age, gender, smoking, level of education, body mass index and physical activity. The benzoyl-DL-arginine-naphthylamide (BANA) test was used to determine the number of periodontal sites with periodontal pathogens. Results: CIMT value was significantly higher in subjects with periodontitis than those without it (0.775 ± 0.268 and 0.683 ± 0.131 mm respectively, p = 0.027). C-reactive protein, leukocyte count and percentage of sites with periodontal pathogens were also significantly higher in subjects with periodontitis. Regression analysis identified age, periodontitis, and smoking as independent predictors of CIMT. Conclusions: These results suggest that untreated periodontitis is associated with early atherosclerotic carotid lesions and higher levels of infl ammatory markers.

Relationship between fluid-attenuated inversion-recovery (FLAIR) signal intensity and inflammatory mediator's levels in the hippocampus of patients with temporal lobe epilepsy and mesial temporal sclerosis / Correlação da intensidade do sinal em FLAIR e os níveis de mediadores inflamatórios no hipocampo de pacientes com epilepsia do lobo temporal e esclerose mesial temporal

We investigated a relationship between the FLAIR signal found in mesial temporal sclerosis (MTS) and inflammation. Twenty nine patients were selected through clinical and MRI analysis and submitted to cortico-amygdalo-hippocampectomy to seizure control. Glutamate, TNFα, IL1, nitric oxide (NO) levels and immunostaining against IL1β and CD45 was performed. Control tissues (n=10) were obtained after autopsy of patients without neurological disorders. The glutamate was decreased in the temporal lobe epilepsy (TLE) -MTS group (p<0.001), suggesting increased release of this neurotransmitter. The IL1β and TNFα were increased in the hippocampus (p<0.05) demonstrating an active inflammatory process. A positive linear correlation between FLAIR signal and NO and IL1β levels and a negative linear correlation between FLAIR signal and glutamate concentration was found. Lymphocytes infiltrates were present in hippocampi of TLE patients. These data showed an association between hippocampal signal alteration and increased inflammatory markers in TLE-MTS.

Este estudo foi delineado para investigar a presença de relação entre a intensidade de sinal em FLAIR e níveis de citocinas, óxido nítrico (NO) e glutamato no hipocampo de pacientes com epilepsia do lobo temporal refratária, associada com esclerose mesial (TLE-MTS). Vinte e nove pacientes foram selecionados através de análise clínica e de ressonância magnética (RM) que foram submetidos a cortico-amigdalo-hipocampectomia para o controle das crises. Os níveis de glutamato foram avaliados por HPLC, as citocinas TNFα e IL1β por ELISA e os níveis de NO via NO system. Avaliamos também por imuno-histoquímica a expressão de IL1β e CD45 em tecidos controles e com esclerose. Tecido controle foi obtido após autópsia de indivíduos mortos sem disfunções inflamatórias e neurológicas (n=10). A concentração de glutamato se mostrou reduzida no tecido TLE-MTS (p<0,001) sugerindo aumento na liberação desse neurotransmissor. TNFα e IL1β também apresentaram níveis elevados no hipocampo dos pacientes (p<0,05), demonstrando um processo inflamatório crônico. Houve uma correlação linear positiva entre a intensidade do sinal em FLAIR e os níveis de NO e IL1β. Em contraste, uma correlação linear negativa foi encontrada entre a intensidade do sinal em FLAIR e níveis de glutamato no hipocampo com esclerose. Infiltrado linfocitário hipocampal também foi visualizado pela imuno-marcação com CD45 em pacientes com TLE-MTS. Esses dados mostraram uma associação entre alteração de sinal na RM e marcadores inflamatórios em pacientes com TLE-MTS.

Changing Patterns of Acute Phase Proteins and Inflammatory Mediators in Experimental Caprine Coccidiosis

This experiment was conducted to assess the changing patterns and relative values of acute phase proteins and inflammatory cytokines in experimental caprine coccidiosis. Eighteen newborn kids were allocated to 3 equal groups. Two groups, A and B, were inoculated with a single dose of 1x10(3) and 1x10(5) sporulated oocysts of Eimeria arloingi, respectively. The third group, C, received distilled water as the control. Blood samples were collected from the jugular vein of each kid in both groups before inoculation and at days 7, 14, 21, 28, 35, and 42 post-inoculation (PI), and the levels of haptoglobin (Hp), serum amyloid A (SAA), TNF-alpha, and IFN-gamma were measured. For histopathological examinations, 2 kids were selected from each group, euthanized, and necropsied on day 42 PI. Mean Hp concentrations in groups A and B (0.34 and 0.68 g/L) at day 7 PI were 3.2 and 6.3 times higher than the levels before inoculation. The mean SAA concentrations in groups A and B (25.6 and 83.5 microg/ml) at day 7 PI were 4.2 and 13.7 times higher than the levels before inoculation. The magnitude and duration of the Hp and SAA responses correlated well with the inoculation doses and the severity of the clinical signs and diarrhea in kids. These results were consistent with the histopathological features, which showed advanced widespread lesions in group B. In both groups, significant correlations were observed for TNF-alpha and IFN-gamma with SAA and Hp, respectively. In conclusion, Hp and SAA can be useful non-specific diagnostic indicators in caprine coccidiosis.

Correlação entre mediadores inflamatórios na secreção nasofaríngea e no soro de crianças com infecção do trato respiratório inferior por vírus sincicial respiratório e a gravidade da doença / Correlation between inflammatory mediators in the nasopharyngeal secretion and in the serum of children with lower respiratory tract infection caused by respiratory syncytial virus and disease severity

OBJETIVO: Avaliar se as concentrações dos mediadores inflamatórios (CCL5, soluble intercellular adhesion molecule type 1 [sICAM-1], TNF-α, IL-6 e IL-10) na secreção nasofaríngea e no soro de crianças com infecção do trato respiratório inferior (ITRI) por vírus sincicial respiratório (VSR) apresentam correlação com os marcadores clínicos de gravidade da doença. MÉTODOS: Entre julho de 2004 e dezembro de 2005, 30 crianças com idade inferior a três meses, diagnosticadas com ITRI por VSR e admitidas em uma UTI neonatal foram incluídas neste estudo. RESULTADOS: Houve uma correlação positiva significante entre a gravidade da doença na admissão hospitalar, determinada por um sistema de escore clínico modificado, e as concentrações de sICAM-1 e de IL-10 na secreção nasofaríngea e de IL-6 no soro dos pacientes. Houve também uma correlação positiva significante entre a concentração de IL-6 no soro e o tempo de oxigenoterapia e a duração da internação. CONCLUSÕES: As concentrações de sICAM-1 e IL-10 na secreção nasofaríngea e de IL-6 no soro determinadas na admissão poderiam ser usadas como marcadores de gravidade da ITRI por VSR. Os níveis de IL-6 determinados no soro na admissão também poderiam ser usados para predizer o prolongamento da oxigenoterapia e da duração da internação.

OBJECTIVE: To determine whether the concentrations of inflammatory mediators (CCL5, soluble intercellular adhesion molecule type 1 [sICAM-1], TNF-α, IL-6 and IL-10) in the nasopharyngeal secretion and in the serum of children with lower respiratory tract infection (LRTI) caused by respiratory syncytial virus (RSV) correlate with the clinical markers of disease severity. METHODS: Between July of 2004 and December of 2005, 30 children less than three months of age, diagnosed with LRTI caused by RSV and admitted to a neonatal ICU, were included in this study. RESULTS: The severity of disease at hospital admission, as determined with a modified clinical scoring system, presented a significant positive correlation with sICAM-1 and IL-10 concentrations in the nasopharyngeal secretion, as well as with IL-6 concentrations in the serum, of the patients. In addition, serum IL-6 concentrations presented a significant positive correlation with the duration of oxygen therapy and with the length of hospital stay. CONCLUSIONS: At hospital admission, the concentrations of sICAM-1 and IL-10 in the nasopharyngeal secretion, as well as the concentration of IL-6 in the serum, could be used as markers of severity in patients with LRTI caused by RSV. The serum levels of IL-6 determined at admission could also be used to predict prolonged oxygen supplementation and hospital stay.

Inflammatory response in a rat model of gastroschisis is associated with an increase of NF-kappaB

Babies with gastroschisis have high morbidity, which is associated with inflammatory bowel injury caused by exposure to amniotic fluid. The objective of this study was to identify components of the inflammatory response in the intestine and liver in an experimental model of gastroschisis in rats. The model was surgically created at 18.5 days of gestation. The fetuses were exposed through a hysterotomy and an incision at the right of the umbilicus was made, exposing the fetal bowel. Then, the fetus was placed back into the uterus until term. The bowel in this model had macro- and microscopic characteristics similar to those observed in gastroschisis. The study was conducted on three groups of 20 fetuses each: gastroschisis, control, and sham fetuses. Fetal body, intestine and liver weights and intestine length were measured. IL-1â, IL-6, IL-10, TNF-á, IFN-ã and NF-kappaB levels were assessed by ELISA. Data were analyzed statistically by ANOVA followed by the Tukey post-test. Gastroschisis fetuses had a decreased intestine length (means ± SD, 125 ± 25 vs 216 ± 13.9; P < 0.005) and increased intestine weight (0.29 ± 0.05 vs 0.24 ± 0.04; P < 0.005). Intestine length correlated with liver weight only in gastroschisis fetuses (Pearson’s correlation coefficient, r = 0.518, P = 0.019). There were no significant differences in the concentrations of IL-1â, TNF-á or IFN-ã in the intestine, whereas the concentration of NF-kappaB was increased in both the intestine and liver of fetuses with gastroschisis. These results show that the inflammatory response in the liver and intestine of the rat model of gastroschisis is accompanied by an increase in the amount of NF-kappaB in the intestine and liver.

Divergent effects of Nitric oxide on airway epithelial cell activation

Nitric oxide (NO*) is a gaseous mediator synthesized by Nitric oxide sinthases. NO* is involved in the modulation of inflammation, but its role in airway inflammation remains controversial. We investigated the role of NO* in the synthesis of the chemok Nes Interleukin-8 and Monocyte Chemotactic Protein-1, and of Intercellular Adhesion Molecule-1 by human airway epithelial cells. normal human bronchial epithelial cells and the bronchial epithelial cell line BEAS-2B were used. Neterleukin-8 (IL-8) and Monocyte Chemotactic Protein-1 (MCP-1) secretion and Intercellular Adhesion Molecule-1 (ICAM-1) expression were measured by ELISA. mRNA was assessed by semiquantitative RTI-PCR. Neterleukin-8 secretion was significantly reduced after 24h incubation with the NO* donor, sodium nitroprusside. The effect was dose-dependent. Similar results were obta Ned with S-Nitroso-N-D,L-penicillam Ne and S-Nitroso-L-glutathione. Inhibition of endogenous NO* with the Nitric oxide synthase inhibitor N-Nitro-L-arg N Ne-methyl-esther caused an increase in IL-8 secretion by lypopolisaccharide- and cytok Ne-stimulated BEAS-2B cells. Sodium nitroprusside also caused a reduction in Monocyte Chemotactic Protein-1 secretion by both cell types. In contrast, Intercellular Adhesion Molecule-1 expression was upregulated by sodium NItroprusside. RTI-PCR results indícate that the modulation of protein levels was paralleled by modification in mRNA levels. NO* has divergent effects on the synthesis of different inflammatory mediators in human bronchial epithelial cells.

Non-asthmatic patients show increased exhaled nitric oxide concentrations

OBJECTIVE: Evaluate whether exhaled nitric oxide may serve as a marker of intraoperative bronchospasm. INTRODUCTION: Intraoperative bronchospasm remains a challenging event during anesthesia. Previous studies in asthmatic patients suggest that exhaled nitric oxide may represent a noninvasive measure of airway inflammation. METHODS: A total of 146,358 anesthesia information forms, which were received during the period from 1999 to 2004, were reviewed. Bronchospasm was registered on 863 forms. From those, three groups were identified: 9 non-asthmatic patients (Bronchospasm group), 12 asthmatics (Asthma group) and 10 subjects with no previous airway disease or symptoms (Control group). All subjects were submitted to exhaled nitric oxide measurements (parts/billion), spirometry and the induced sputum test. The data was compared by ANOVA followed by the Tukey test and Kruskal-Wallis followed by Dunn's test. RESULTS: The normal lung function test results for the Bronchospasm group were different from those of the asthma group (p <0.05). The median percentage of eosinophils in induced sputum was higher for the Asthma [2.46 (0.45-6.83)] compared with either the Bronchospasm [0.55 (0-1.26)] or the Control group [0.0 (0)] (p <0.05); exhaled nitric oxide followed a similar pattern for the Asthma [81.55 (57.6-86.85)], Bronchospasm [46.2 (42.0 -62.6] and Control group [18.7 (16.0-24.7)] (p< 0.05). CONCLUSIONS: Non-asthmatic patients with intraoperative bronchospasm detected during anesthesia and endotracheal intubation showed increased expired nitric oxide.

Capacidade dos biomarcadores inflamatórios em predizer a síndrome metabólica / Inflammation biomarkers capacity in predicting the metabolic syndrome

O processo inflamatório é o elo entre a síndrome metabólica e as doenças cardiovasculares. Para medir o grau da inflamação subclínica, vários biomarcadores inflamatórios têm sido propostos. Este trabalho tem como objetivo revisar as recentes pesquisas das associações entre os biomarcadores inflamatórios e a síndrome metabólica, bem como a capacidade daqueles em predizer a síndrome metabólica. Estes biomarcadores incluem as citocinas pró-inflamatórias, citocinas antiinflamatórias, adipocinas, chemocinas, marcadores de inflamação derivados de hepatócitos, marcadores de conseqüência da inflamação e enzimas. Com esta revisão pode-se integrar o novo conhecimento referente às interações possíveis de mediadores inflamatórios com a síndrome metabólica, visto que estes biomarcadores desempenham vários papéis e seguem diversos caminhos metabólicos.

The inflammatory process is the link between metabolic syndrome and cardiovascular diseases. To measure the degree of subclinical inflammation some inflammatory biomarkers have been considered. This work reviews the recent researches of the associations between inflammatory biomarkers and metabolic syndrome, as well as the capacity in predicting the metabolic syndrome. These biomarkers include pro-inflammatory cytokines, anti-inflammatory cytokines, adipokines, chemokines, inflammation markers derived from hepatocites, the consequence markers of inflammation and enzymes. This review integrates the new knowledge of inflammatory mediators interactions with metabolic syndrome, since these biomarkers play different roles and follow diverse metabolic ways.

Necrotising enterocolitis: the state of the science.

Necrotizing enterocolitis is the most common gastrointestinal emergency of the neonate, affecting 5-10% of infants, yet the pathogenesis remains unclear. Widely accepted risk factors include prematurity, enteral feeds, bacterial colonization and mucosal injury. How these or other yet identified factors come together to create the classic clinical and pathologic features is the subject of much research. The activation of the cytokine cascade, in part by bacterial ligands, appears to play a key role in mucosal injury. Two mediators that may also contribute are platelet activating factor and intestinal toll-like receptors. Short chain fatty acids, the products of bacterial fermentation of carbohydrates, have been thought to cause mucosal injury. Overgrowth of pathogenic bacteria in the face of a decreased commensal population may play a key role. A current focus of clinical research involves probiotics, enterally fed forms of commsenal bacteria. This may set the stage for a healthier intestinal ecosystem and possibly, decreased risk of NEC.

Effect of hyperthermia on experimental acute pancreatitis

BACKGROUD: Recent studies indicate that hyperthermia can change inflammatory mechanisms and protect experimental animals from deleterious effects of secretagogue-induced acute pancreatitis AIM: To evaluate the effects of hyperthermia post-treatment on cerulein-induced acute pancreatitis in rats METHODS: Twenty animals were divided in two groups: group I (n = 10), rats with cerulein-induced acute pancreatitis undergone hyperthermia, and group II (n = 10), animals with cerulein-induced acute pancreatitis that were kept normothermic. In all groups, amylase serum levels, histologic damage, vascular permeability and pancreatic water content were assessed. Acute pancreatitis was induced by administration of two cerulein injections (20 mcg/kg). A single dose of Evans' blue dye was administered along with the second dose of cerulein. All animals also received a subcutaneous injection of saline solution. After this process, animals undergone hyperthermia were heated in a cage with two 100 W lamps. Body temperature was increased to 39.5°C and maintained at that level for 45 minutes. Normothermia rats were kept at room temperature in a second cage RESULTS: Control animals had typical edema, serum amylase activity and morphologic changes of this acute pancreatitis model. Hyperthermia post-treatment ameliorated the pancreatic edema, whereas the histologic damage and the serum amylase level remained unchanged CONCLUSIONS: The findings suggest a beneficial effect of the thermal stress on inflammatory edema in experimental acute pancreatitis.

RACIONAL: Estudos recentes indicam que a hipertermia pode modificar mecanismos inflamatórios e proteger animais experimentais dos efeitos deletérios da pancreatite aguda induzida por secretagogos OBJETIVO: Avaliar a eficácia da hipertermia como tratamento da pancreatite aguda induzida por ceruleína em ratos MÉTODOS: Vinte animais foram divididos em dois grupos: grupo I (n = 10), ratos com pancreatite aguda induzida por ceruleína e submetidos a hipertermia, e grupo II (n = 10), animais com pancreatite aguda induzida por ceruleína mantidos em normotermia. Em todos os grupos foram medidos níveis séricos de amilase, histologia, permeabilidade vascular e conteúdo de água do pâncreas. A pancreatite aguda foi induzida através da administração de duas injeções de ceruleína (20 mcg/ kg). Dose única do corante azul de Evans foi administrada juntamente com a segunda injeção de ceruleína. Todos os animais também receberam 5 mL de solução salina subcutânea. Após a indução, os animais do grupo hipertérmico foram aquecidos com duas lâmpadas de 100 W em gaiola parcialmente isolada. A temperatura corporal foi aumentada para 39,5°C e mantida neste nível por 45 minutos. Os animais controle foram mantidos em uma segunda gaiola em temperatura ambiente RESULTADOS: Os animais controle tiveram edema, danos histológicos e níveis de amilase típicos do modelo de pancreatite aguda leve com ceruleína. O tratamento com hipertermia melhorou o edema pancreático porém não teve efeito nos nível séricos de amilase e no dano histológico pancreático CONCLUSÕES: Os resultados sugerem efeito benéfico da hipertermia no edema inflamatório da pancreatite aguda leve experimental.

Análise de componentes celulares, mediadores inflamatórios e apoptose no processo de rejeição ao enxerto no transplanted e pâncreas-rim / Analysis of the celular components, inflammatory mediators and aoptosis in the process of allograft rejection in pancreas-kidney transplantation

O objetivo do presente estudo, prospectivo e randomizado, é avaliar as respostas hemodinâmicas, gasométricas e imunomoleculares em pacientes submetidos a cirurgia de revascularização miocárdica utilizando-se, em um dos dois grupos, a infusão de azul de metileno durante o período intra-operatório. Ocorreram alterações estatisticamente significativas na resistência vascular sistêmica, na pressão diastólica arterial sistêmica, em parte das citocinas estudadas e no óxido nítrico. A análise dos resultados mostra que no grupo de pacientes que utilizou azul de metileno houve resistência vascular sistêmica mais alta, maiores concentrações das citocinas anti-inflamatórias e menores de óxido nítrico. Houve, no entanto, neste grupo, valores mais altos de IL-8 mas não ocorreram alterações estatisticamente significativas nas dosagens de IL-6. As medidas gasométricas foram semelhantes em ambos os grupos /The objectiv of the present study, prospective and randomized, is to evaluate the hemodynamic, gasometric and immunomolecular response in patients submited to coronary artery bypass surgery with the use, in one of the two groups, of methylene blue infusion during the intraoperative period. It was observed that stastiscally significant diffences ocurred in systemic vascular resistance, dyastolic systemic arterial pressure and in part of the studied cytokines and in the nitric oxide levels. The analysis of the results shows that in the group of patients with methylene blue, systemic vascular resistance was higher, as well as the levels of the anti-inflamatory cytokines and less nitric oxide concentration. Neverthless, there was in this group, higher valvues of IL-8 but there was no difference in IL-6 levels. The gasometric study showed no difference between the groups...

Inflammatory mediators in nasal lavage among school-age children from urban and rural areas in São Paulo, Brazil

CONTEXTO: Alguns estudos demonstram que o processo inflamatório nas vias aéreas nasais poderia refletir ou mesmo afetar as vias aéreas inferiores. Decidimos avaliar indiretamente o estado inflamatório das vias aéreas nasais de dois grupos de escolares com diferente sensibilização aos aeroalérgenos mais comuns. OBJETIVO: Comparar a atividade inflamatória nas vias aéreas nasais, através da determinação de mediadores inflamatórios no lavado nasal em duas populações distintas de crianças em idade escolar. TIPO DE ESTUDO: Estudo transversal. LOCAL: O estudo foi realizado em duas escolas públicas de ensino fundamental, uma em zona urbana e outra em zona rural, no Estado de São Paulo. MÉTODOS: Foram constituídos dois grupos de 40 escolares que apresentam diferentes taxas de sensibilização a aeroalérgenos comuns. Amostras do lavado nasal foram colhidas para determinação de proteína catiônica eosinofílica (ECP) e triptase. Testes não-paramétricos foram usados na análise estatística. RESULTADOS: Níveis significativamente maiores de proteína catiônica eosinofílica foram encontrados nos estudantes da área urbana (p < 0,05). Não houve diferença estatística nos níveis de triptase entre os dois grupos. Observou-se ainda que, na área urbana, as crianças sensibilizadas aos aeroalérgenos apresentaram maiores concentrações de proteína catiônica eosinofílica, o que não foi observado nas crianças da zona rural. DISCUSSAO: A ausência de atividade de mastócitos e a degranulação aumentada de eosinófilos revelaram uma inflamação crônica nas vias aéreas das crianças estudadas. A maior atividade de eosinófilos na zona urbana, coincidindo com a maior sensibilização aos aeroalérgenos, sugere que deve haver algum fator a mais na área urbana que modula a resposta das vias aéreas influenciando a ativação das células inflamatórias locais. CONCLUSAO: Nossos achados não mostraram diferenças nos níveis de triptase no lavado nasal entre os dois grupos estudados. Por outro lado, as crianças da area urbana apresentaram maiores concentrações de proteína catiônica eosinofílica do que aquelas da zona rural. Observamos ainda que, na area urbana, as crianças sensibilizadas por aeroalérgenos apresentaram maiores concentrações de proteína catiônica eosinofílica do que aquelas não sensibilizadas, enquanto esta diferença não foi observada nas crianças da area rural.

Effects of Russell's viper venom on mediator production in cultured human umbilical vein endothelial cells.

Hemodynamic alterations in Russell's viper envenomation are the result of interactions of various vasoactive mediators and perhaps proinflammatory cytokines. Since vascular endothelium is likely to be exposed to high concentrations of the venom and the endothelial cell itself not only plays an important role in the physiologic control of the circulation, but also play a role in inflammation with the synthesis and secretion of proinflammatory cytokines. It was therefore, the objective of this study to determine the effects of Russell's viper venom (RVV) on proinflammatory cytokine production by cultured human umbilical vein endothelial cells (HUVEC) and the release of endothelium-derived substances. Endothelial cells were isolated from freshly obtained human umbilical cord vein and grown in tissue culture to confluence as a homogeneous population. Cells were then incubated at 37 degrees C under 5 per cent CO2 with RVV (0.2, 1.0, 5.0, and 25.0 microg/ml) or lipopolysaccharide (LPS, 10 microg/ml) for 3, 6, 12 and 24 hours. After an indicated time, the levels of endothelin-1 (ET-1); 6-keto-PGF1alpha (a stable metabolite of PGI2) tumor necrosis factor-alpha (TNF-alpha); interleukin-1beta (IL-1beta); and interleukin-6 (IL-6) in supernatants were measured by using ELISA or EIA. The effect of RVV or LPS on cell viability was also measured using MIT assay. The results showed copious amounts of ET-1 production irrespectively with the presence of RVV or LPS. Whereas, production of PGI2 (measured as 6-keto-PGF1alpha, a stable metabolite) was increased significantly higher in the RVV- and LPS-treated EC than in the control EC. However, TNF-alpha and IL-6 productions were not different among these groups. The levels of IL-1beta were very low, although IL-1beta was detectable in the group treated with RVV at a concentration of 25.0 microg/ml. In conclusion, RVV upto 25 microg/ml stimulated PGI2 production by cultured HUVEC. This effect was unlikely related to production of proinflammatory cytokines since LPS or RVV is not sufficient per se to elevate a substantial amount of EC-derived cytokines. The higher amount of IL-6 compared to TNF-alpha and IL-1beta may be produced through other pathways apart from production via a cascade of cytokines. This is the first report showing that RVV up to 25 microg/ml has no effect on prominent proinflammatory cytokine production by HUVEC. However, in blood circulation, the major source of cytokines production is monocyte-macrophage lineage cell. Thus, RVV in blood circulation may activate the production of proinflammatory cytokines mainly from those cells and subsequently induce toxicity.

The significance and technical aspects of quantitative measurements of inflammatory mediators in allergic rhinitis.

The pathophysiology of allergic rhinitis induced by various inhalant allergens through an IgE mediated mechanism, has been well demonstrated. The participation of many important inflammatory cells and mediators released by these cells in the human nasal allergic reaction provides insight into the relationship between the responsiveness to allergen exposure and nasal symptoms of allergic rhinitis. This paper summarizes our previous studies on some important mediators in the nasal secretions of atopic patients during different phases after nasal allergen challenge and during natural allergen exposure. The microsuction technique proves to be an especially useful and reliable nasal sampling method permitting quantitative analysis of important mediators such as histamine, tryptase, leukotriene C4 and eosinophil cationic protein in nasal secretions. The measurement of these mediators during allergic reactions provides accurate data on the activity of some important inflammatory cells (i.e., mast cells, basophils, and eosinophils) and their responses to therapy.

Analise de mediadores pro-inflamatorios na resposta alergica a antigenos de acaro de poeira domestica em camundongos / Pro-inflamatory mediators analisys in the alergic response to mite antigens of domestic dust in rats

Ha um grande contigente de pacientes com rinite alergica na populacao que nao se beneficiam com os tratamentos convencionais, tornando seu tratamento um desafio para o corpo clinico. A fim de descrever os mecanismos imunes e inflamatorios que envolvem o pulmao na broncoprovocacao por Dhermatophagoides pteronyssinus, o presente estudo utilizou de metodos de citometria de fluxo, cultura de celulas e e contagem total e diferencial de celulas e dosagem de NO para a analise do lavado bronco alveolar...