Myeloperoxidase and adenosine-deaminase levels in the pleural fluid leakage induced by carrageenan in the mouse model of pleurisy.

BACKGROUND: Although myeloperoxidase (MPO) and adenosine-deaminase (ADA) levels are markers of activated leukocytes, both enzymes have not been currently addressed in inflammation models. AIMS: This study evaluates whether the concentrations of these enzymes are significantly correlated with the content of leukocytes in a pleurisy model. METHODS: The pleurisy was induced by carrageenan (1%) in mice, and the parameters analyzed 4 and 48 h after. RESULTS: After the induction of inflammation (4h), MPO and ADA levels peaked in parallel to neutrophils (p<0.01). Regarding the second phase of pleurisy (48 h), the highest concentrations of ADA were detected in parallel to the highest levels of mononuclears (p<0.01). At this time, MPO levels and neutrophils remained elevated, although at lower levels than those found at 4 h. A significant positive correlation was found among neutrophiLs and MPO, and mononuclears and ADA (p<0.01). CONCLUSIONS: These findings support the evidence that both enzymes are markers of the inflammatory process, and provide new tools for a better understanding of the immunoregulatory pathways that occur in inflammation

Mediadores e mecanismos envolvidos na pleurisia induzida pela carragenina em camundongos /

Tese (Doutorado) - Universidade Federal de Santa Catarina, Centro de Ciências Biológicas.O modelo da [pleurisia] induzida pela [carragenina] (1%), apresenta resposta inflamatória bifásica (4 e 48 h) na cavidade pleural de [camundongos]. A L-NOARG, o HOE 140, o NPC 17731, a diacereína, a indometacina, o meloxicam, a nabumetona e a dexametasona administrados 30 min antes da [carragenina], inibiram a migração celular e os níveis da [mieloperoxidase] na primeira fase (4 h) desta resposta inflamatória. Todas as drogas, com exceção da diacereína, inibiram a exsudação. Além disso, a diacereína, a nabumetona, a L-NOARG e a L-NAME inibiram também os níveis de [nitrito/nitrato]. Em relação à segunda fase (48 h), observou-se que a dexametasona e a diacereína, inibiram os leucócitos, enquanto que a terfenadina reduziu apenas a exsudação. Nenhuma das drogas estudadas reduziu os níveis da [mieloperoxidase] ou do [nitrito/nitrato]. O NPC 17731, o HOE 140 e o NPC 18884 inibiram tanto a exsudação como a migração celular induzida pela [carragenina], 4 h. Neste modelo experimental observa-se a participação de [citocinas] com efeitos pró-inflamatórios (TNFa, IL-1b, IL-8) e antiinflamatórios (IL-6, IL-10), com base nos resultados de que tanto anticorpos-policlonais como citocinas-recombinantes, administrados diretamente na cavidade pleural juntamente com a [carragenina], reproduziram parcialmente seus efeitos pró-inflamatórios ou antiinflamatórios. Neste modelo de inflamação, o PDTC e a sulfasalazina (inibidores do fator de transcrição nuclear [NF-kB]), foram eficazes em inibir a resposta inflamatória induzida pela [carragenina] (4 h). Além disso, a associação de doses subliminares de drogas antiinflamatórias e dos inibidores (PDTC ou sulfasalazina) que isoladamente não foram eficazes, quando associados, mostraram efeito antiinflamatório sinergístico, inibindo tanto a migração celular como a exsudação na [pleurisia] induzida pela [carragenina] (4 h). Nestes experimentos observou-se também aumento da expressão de células [CD18] positivas, na cavidade pleural e no pulmão, na primeira fase (4 h) da resposta inflamatória induzida pela [carragenina]. Além disso, no modelo experimental estudado, não se observou resposta inflamatória sistêmica caracterizada por aumento da permeabilidade vascular nos pulmões e no baço, em que se pese o aumento da molécula de adesão [CD18] em células pulmonares

Aufarbeitung der Schulmusikerziehung der DDR - eine Bilanz nach zehn Jahren. Wege und Perspektiven

An drei Befunde in Sachen Aufarbeitung der Musikpädagogik in der DDR schließt der Autor seine Bilanz an. Er will in diesem Beitrag die Versuche wissenschaftlichen und nicht-wissenschaftlichen Nachdenkens über die schulmusikalische Vergangenheit in der DDR benennen und ordnen, kritisch würdigen, auf Defizite aufmerksam machen und auf Perspektiven hinweisen. (DIPF/Orig.

Analysis of local and systemic inflammatory responses induced by polymicrobial peritonitis in mice.

BACKGROUND: Abdominal sepsis induces a local production of proinflammatory mediators that may trigger both septic shock and organ-system dysfunction. AIMS: The present study analyzed exudation, cell migration, and CD11a and CD18 subset cells of both local and systemic responses induced by fecal peritonitis in mice. METHODS: Animals were anesthetized and, after performing a midline incision in the abdomen, the cecum was ligated and punctured twice with a needle. Sham-operated animals were included. Some groups were previously treated with Evans blue dye (intravenously) to further evaluate the amount of tissue and abdominal cavity leakages. RESULTS: Fecal peritonitis triggered a local inflammatory reaction with an increased number of leukocytes and exudation between 6 and 48 h (p < 0.01). Although CD11a/CD18-positive cells in the abdomen peaked after 24h, a significant decrease of them was detected after 48 h (p < 0.05). At the studied period of time (6-48 h), different degrees of exudation in several organs occurred, whereas a significant late recruitment (24 h) of CD11a/CD18 cells into the lungs was observed. CONCLUSIONS: In this model, cell migration and exudation at the site of injury occurred in parallel. However, in the lungs, the recruitment of leukocytes that express CD11a/CD18 adhesion molecules constitutes a non-dependent event in relation to fluid leakage accumulation at this site

Effects of methotrexate upon inflammatory parameters induced by carrageenan in the mouse model of pleurisy.

BACKGROUND: The model of pleurisy induced by carrageenan exhibits a biphasic response (4 and 48 h) and permits the quantification of exudate, cell migration and certain enzymes such as myeloperoxidase (MPO) and adenosine-deaminase (ADA) that are markers of activated leukocytes. AIMS: The present study evaluates whether there exists, in the pleurisy model, a significant inhibition of ADA and MPO enzymes, leukocyte kinetics and other markers of inflammation [nitric oxide (NO) levels, exudation] caused by methotrexate treatment by the intraperitoneal (i.p.) route. METHODS: The pleurisy was induced by carrageenan (1%) in mice, and the parameters were analyzed 4 and 48 h after. RESULTS: After the induction of inflammation (4 h), methotrexate (20 mg/kg, i.p., 24 h before pleurisy induction) inhibited the leukocyte infiltration (p < 0.05), NO levels and MPO activity (p < 0.01), but not ADA activity and fluid leakage (p > 0.05). Regarding the second phase of pleurisy (48 h), methotrexate (40 mg/kg, i.p., 0.5 h before pleurisy induction) inhibited the leukocyte infiltration (p < 0.05), fluid leakage, NO levels (p < 0.01), and ADA and MPO activity (p < 0.05). CONCLUSIONS: These findings support the evidence that the acute administration of methotrexate has an important systemic anti-inflammatory activity in the studied inflammatory model. This effect was due to a significant inhibition on both neutrophil and mononuclear cells, being less marked in relation to exudation 48 h after. In relation to the enzymes studied and to NO levels, the findings support the evidence that methotrexate inhibits both enzymes (MPO and ADA) from leukocytes at the site of injury, thus reflecting the activation of both neutrophils and lymphocytes, respectively. Furthermore, the inhibiting effect on NO in both phases of pleurisy induced by carrageenan (4 and 48 h) indicates that methotrexate acts on constitutive and/or inducible NO synthases by means of different cells of the pleural cavity

Involvement of steroids in anti-inflammatory effects of PK11195 in a murine model of pleurisy.

BACKGROUND: Studies on peripheral benzodiazepine receptor function have yielded a diverse list of activities of which the anti-inflammatory effects need to be further examined. AIMS: To evaluate the role of steroids, nitric oxide and adenosine-deaminase in the anti-inflammatory effect of PK11195. METHODS: Pleurisy was induced by intrapleural injection of carrageenan in mice pre-treated or not with PK11195. Leukocytes, exudation, adenosine-deaminase (ADA) activity and nitric oxide (NO) level were measured. Steroid involvement was evaluated by pre-treatment with D,L-aminogluthetimide before PK11195. RESULTS: Leukocytes, exudation and NO levels were reduced by PK11195 in the early (4 h) phase. In the late (48 h) phase, PK11195 decreased leukocytes and ADA activity. D,L-aminogluthetimide reversed the effect of PK11195 on exudate (4 h), as well as total and differential leukocytes and NO levels (48 h). CONCLUSIONS: Steroids, NO and ADA are implicated in the anti-inflammatory action of PK11195

Activation of Human Neutrophils by the Anti-Inflammatory Mediator Esenbeckia leiocarpa Leads to Atypical Apoptosis

Despite the fact that Esenbeckia leiocarpa, a Brazilian plant, possesses potential anti-inflammatory properties, its effect in neutrophils, key players in inflammation, has never been investigated. In this study, a crude hydroalcoholic extract (CHE) was used to evaluate the potential toxic or agonistic effect of E. leiocarpa in human neutrophils. At a noncytotoxic concentration of 500 μg/mL, CHE increased actin polymerization and cell signaling events, especially p38 MAPK. Its modulatory activity on neutrophil cell apoptosis was investigated by cytology and by flow cytometry and, although CHE increased the apoptotic rate (by cytology) and increased annexin-V binding, it did not, unexpectedly, increase CD16 shedding. CHE increased the degradation of the cytoskeletal proteins gelsolin and paxillin but, surprisingly, not of vimentin. The proapoptotic activity of CHE was reversed by a pan-caspase inhibitor but not by a p38 inhibitor. We conclude that CHE is a novel human neutrophil agonist that induces apoptosis by a caspase-dependent and p38-independent mechanism in an atypical fashion based on its lack of effect on CD16 shedding and vimentin degradation. Since the resolution of inflammation occurs by elimination of apoptotic neutrophils, the ability of CHE to induce neutrophil apoptosis correlates well with its anti-inflammatory properties, as previously reported

Tumour necrosis factor-alpha, interleukin-2 soluble receptor and different inflammatory parameters in patients with rheumatoid arthritis.

BACKGROUND AND AIMS: Although the participation of cytokines in the pathogenesis of rheumatoid arthritis (RA) seems to be unequivocal, their relationship with current serum markers of this disease is not clear. The present study analyses whether there is any correlation between the levels of tumour necrosis factor-alpha (TNF-alpha), interleukin-2 soluble receptor (sIL-2R) and the concentrations of C-reactive protein (CRP), erythrocyte sedimentation rate (ESR) and beta(2)-microglobulin in a group of 21 patients with RA, all rheumatoid factor positive. METHODS: The levels of TNF-alpha and sIL-2R were analysed in association with other parameters of inflammation (ESR, CRP and beta(2)-microglobulin). RESULTS: In comparison with the control group, RA patients presented high median levels of both cytokines, TNF-alpha (6.4 pg/ml) and sIL-2R (56 pmol/L), as well as of ESR (34 mm/h), CRP (0.9 mg/dl) and beta(2)-microglobulin (1.6 mg/dl) (p < 0.01). However, only ESR levels in the RA group significantly differ from the control group (p < 0.01). No correlation was found between the inflammatory parameters. CONCLUSIONS: These results suggested that TNF-alpha and slL-2R levels are up-regulated in RA patients but did not significantly differ from the control group. Due to the chronic course of this disease, other inflammatory markers must be identified in order to provide early therapeutic strategies to these patients

Diagnostic Yield and Benefits of Whole Exome Sequencing in CAKUT Patients Diagnosed in the First Thousand Days of Life

Infancy; Reverse phenotyping; Whole exome sequencingInfancia; Fenotipado inverso; Secuenciación del exoma completoInfància; Fenotipat invers; Seqüenciació de l'exoma completIntroduction Congenital anomalies of the kidney and urinary tract (CAKUT) are the predominant cause of chronic kidney disease (CKD) and the need for kidney replacement therapy (KRT) in children. Although more than 60 genes are known to cause CAKUT if mutated, genetic etiology is detected, on average, in only 16% of unselected CAKUT cases, making genetic testing unproductive. Methods Whole exome sequencing (WES) was performed in 100 patients with CAKUT diagnosed in the first 1000 days of life with CKD stages 1 to 5D/T. Variants in 58 established CAKUT-associated genes were extracted, classified according to the American College of Medical Genetics and Genomics guidelines, and their translational value was assessed. Results In 25% of these mostly sporadic patients with CAKUT, a rare likely pathogenic or pathogenic variant was identified in 1 or 2 of 15 CAKUT-associated genes, including GATA3, HNF1B, LIFR, PAX2, SALL1, and TBC1D1. Of the 27 variants detected, 52% were loss-of-function and 18.5% de novo variants. The diagnostic yield was significantly higher in patients requiring KRT before 3 years of age (43%, odds ratio 2.95) and in patients with extrarenal features (41%, odds ratio 3.5) compared with patients lacking these criteria. Considering that all affected genes were previously associated with extrarenal complications, including treatable conditions, such as diabetes, hyperuricemia, hypomagnesemia, and hypoparathyroidism, the genetic diagnosis allowed preventive measures and/or early treatment in 25% of patients. Conclusion WES offers significant advantages for the diagnosis and management of patients with CAKUT diagnosed before 3 years of age, especially in patients who require KRT or have extrarenal anomalies.This work was supported by grants from the Else Kröner-Fresenius-Stiftung (2018_Kolleg.12, Clinician Scientist Program TITUS at Hannover Medical School to LW) and the Deutsche Forschungsgemeinschaft (MA 9606/1-1 to HM, and KO 5614/2-1 to AC and RGW)