Severe iron-deficiency anemia as initial manifestation of pulmonary hemosiderosis in a child / Anemia ferropriva grave como manifestaçao inicial de hemossiderose pulmonar em criança

ABSTRACT Idiopathic pulmonary hemosiderosis is a potentially fatal disease that results from episodes of alveolar hemorrhage of unknown origin. The clinical spectrum is varied, and anemia may constitute the only manifestation of illness, preceding other signs and symptoms by several months. We present the case of a 4 year-old child presenting with fever, vomiting and prostration, associated with pallor. He had microcytic and hypochromic anemia refractory to iron therapy. Gastrointestinal bleeding was ruled out after negative extensive etiological investigation. Subsequently, pulmonary infiltrates suggestive of alveolar hemorrhage were observed in the chest radiography. The cytological exam of the bronchoalveolar lavage showed hemosiderin-laden macrophages. After the etiological study, the diagnosis of idiopathic pulmonary hemosiderosis was made by exclusion. He was initiated on corticosteroid therapy, later associated to an immunosuppressive agent, with subsequent correction of anemia and of the radiological pattern. The patient is currently asymptomatic.

RESUMO A hemossiderose pulmonar idiopática é uma doença potencialmente fatal que cursa com episódios de hemorragia alveolar de etiologia desconhecida. As manifestações clínicas são variadas, e a anemia pode constituir o único sinal de doença, precedendo em vários meses os outros sinais e sintomas. Apresenta-se o caso de criança de 4 anos, com febre, vômitos e prostração, associados à palidez. Apresentava anemia microcítica e hipocrômica, refratária à terapêutica com ferro. A hipótese diagnóstica de sangramento gastrintestinal foi excluída, após investigação etiológica extensa, inconclusiva. Posteriormente, em radiografia torácica, foram observados infiltrados sugestivos de hemorragia alveolar. O exame citológico do lavado broncoalveolar mostrou macrófagos com depósitos de hemossiderina. Após estudo etiológico, assumiu-se, por exclusão, o diagnóstico de hemossiderose pulmonar idiopática. Foi iniciada terapêutica com corticoides, associada posteriormente a imunossupressor, com correção subsequente da anemia e do padrão radiológico, encontrando-se, atualmente, assintomático.

Citologia dos lavados traqueobrônquico (LTB) e broncoalveolar (LBA) de bezerros holandeses sadios durante o primeiro mês de vida / Cytology of tracheobronchial and bronchoalveolar lavage in healthy Holteins calves during the first month of life

O período neonatal dos bezerros é um momento crítico para adaptação do recém-nascido à vida extra uterina e o sistema respiratório, um dos mais exigidos funcionalmente, é frequentemente afetado por enfermidades, redundando no prejuízo direto da sua função e acarretando perdas econômicas importantes na pecuária. O ponto básico para reduzir estas perdas, é representado pela adequada avaliação clínica dos neonatos, todavia o diagnóstico baseado exclusivamente no exame físico é muito difícil de ser estabelecido. O uso de exames complementares como a citologia do trato respiratório torna-se uma ferramenta diagnóstica importante nestes casos, porém faz-se necessário, padronizar seus achados frente às diferentes técnicas empregadas para a sua obtenção. Assim, o presente estudo propôs-se acompanhar as variações dos constituintes celulares da região traqueobrônquica e broncoalveolar obtidos por lavados respiratórios pelos métodos de traqueocentese e por colheita nasotraqueal respectivamente, durante o primeiro mês de vida de bezerros sadios. Observou-se alteração no quadro citológico ao longo do tempo, quando a região traqueobrônquica foi lavada, expresso por diminuição da porcentagem de macrófagos alveolares, com aumento de neutrófilos, possivelmente, por maior irritação local provocada pela técnica, que se repetiu sequencialmente e/ou por maior estimulo de microorganismos inalados depositados nesta região. Na região broncoalveolar, não encontraram-se variações nos constituintes celulares em função do tempo. Os resultados permitiram a conclusão que a população celular da região traqueobrônquica modificou-se ao longo das semanas de vida dos bezerros, possivelmente pela técnica empregada e/ou fisiologia normal da região, sendo representadas por maiores magnitudes de neutrófilos. De modo diverso, na região broncolaveolar, as células evidenciaram um comportamento estável durante o primeiro mês de vida dos bezerros neonatos, apresentando predomínio numérico dos macrófagos alveolares.

The neonatal calf is a critical moment for adaptation of the newborn to extra uterine life. The respiratory tract is functionally very demanded and often affected by disease, resulting in direct loss of their function and causing serious economic losses in livestock. The basic point to reduce these losses is appropriate clinical evaluation of neonates; but the diagnosis based solely in physical examination is very difficult to establish. The use of complementary analysis such cytology of the respiratory tract becomes an important diagnostic tool; however their findings must be standardized in the face of different techniques employed. This research studied the dynamics of the cellularity of the bronchoalveolar and tracheobronchial region obtained through lung lavage harvested by nasotracheal catheterization technique and tracheocenthesis respectively, during the first month of life of healthy calves. The tracheobronchial cytology was influenced by the time, showing decreased number of alveolar macrophages and greater number of neutrophils, possibly increased by local irritation caused by the technique, which was repeated sequentially, and/or through greater stimulation of inhaled microorganisms deposited in this region. In the bronchoalveolar region no variation in the cellular constituents in function of time was found. The results allowed the conclusion the cell population of the tracheobronchial region has changed over the week-old calves, possibly due to the technique used and/or to the normal region physiology, represented by higher magnitudes of neutrophils. Otherwise, the cells of the broncholaveolar region showed a stable behavior during the first month of life of newborn calves, presenting numerical predominance of alveolar macrophages.

Morphological analyses of caseous necrosis correlated to CD68+ and CD8+ cells in biopsies of tuberculous pleurisy / Análisis morfológico de la necrosis caseosa y su correlación con células CD68+ y CD8+ en biopsias de pleuritis tuberculosa

Tuberculosis is the most prevalent infectious disease in the world. Granuloma formation and caseous necrosis are hallmarks of M. tuberculosis infection and they represent the protective and inflammatory reactions in the infected tissues. The molecular mechanisms that mediate granuloma necrosis are still not well understood. Objectives: To immunolocalize and correlate the amounts of CD68+ macrophages and CD8+ lymphocytes to caseous necrosis extension in granulomas of tuberculous pleurisy. Methods: The study is a retrospective analysis of 30 pleural biopsies with histopathological diagnosis of chronic granulomatous pleurisy with caseous necrosis. These biopsies were classified according to necrosis intensity as minimal (N1), moderate (N2) and intense (N3). The number of granulomas was also observed and categorized as G1 (1 to 4 granulomas per section), G2 (5 to 8 granulomas per section), and G3 (more than 8 granulomas per section). Results: The means of CD68+ cells counts per mm² in N1, N2 and N3 categories of necrosis were 1,287 +/- 254, 1086 +/- 181 and 930 +/- 115 respectively. The means for CD8+ cells were 483.7 +/- 396, 366.3 +/- 43 and 558 +/- 53 cells per mm² in N1, N2 and N3 respectively. Conclusions: There were no significant statistical correlations between necrosis extension and cell counts. In analyzed biopsies, the number of CD68+ cells was significantly higher than the number of CD8+ cells.

La tuberculosis es una de las enfermedades más prevalentes en el mundo. La formación del granuloma junto con la necrosis caseosa son características propias de la infección por M. tuberculosis y representan reacciones inflamatorias y protectoras en los tejidos infectados. No se conocen bien los mecanismos moleculares que median la necrosis en el granuloma. Los objetivos fueron inmunolocalizar y correlacionar la cantidad de macrófagos CD68+ y linfocitos CD8+ con la extensión de la necrosis caseosa, en los granulomas de tuberculosis pleural. Análisis retrospectivo que incluyeron 30 biópsias con diagnóstico histopatológico de tuberculosis pleural granulomatosa crónica con necrosis caseosa. Estas biópsias fueron clasificadas según la intensidad de necrosis como mínima (N1), moderada (N2) e intensa (N3). También se determinó el número de granulomas, que fueron clasificados como G1 (1 a4 granulomas por sección), G2 (5 a 8 granulomas por sección), y G3 (más de 8 granulomas por sección). La cuantificación de células CD68+ por mm² en las categorías N1, N2 y N3 de necrosis fue de 1,287 +/- 254; 1086 +/-181 y 930 +/- 115, respectivamente. La cuantificación de las células CD68+ fue de 483,7 +/- 396; 366,3 +/- 43 y 558 +/- 53 células por mm² para N1, N2 y N3, respectivamente. No hubo correlación estadísticamente significativa entre la extensión de la necrosis y la cuantificación celular. El número de células CD68+ fue significativamente mayor que el número de células CD8+ en las biópsias analizadas.

In vitro potential cytotoxicity of an adhesive system to alveolar macrophages

The purpose of this study was to evaluate the potential cytotoxicity of Adper Single Bond 2 (SB) simplified etch-and-rinse adhesive system in alveolar macrophage cultures, as a function of the post-polymerization time and duration of immersion in the culture medium for preparation of extracts, by observing the levels of nitric oxide (NO) release and cell survival rate (MTT assay). Wistar rat alveolar macrophages were exposed to 200 μL of extracts obtained from 24- or 72-h immersion of adhesive samples in culture medium (RPMI), immediately or 24 h after polymerization. Fresh RPMI and E. coli lipopolysaccharides were used as negative and positive controls, respectively. The cells were placed in a humidified incubator for 24 h. The results were analyzed by the Student's-t test (α=5 percent). The amount of NO produced and viable cells were significantly different (p<0.05) between the experimental and the control groups, showing that, irrespective of the post-polymerization time and duration of immersion in the culture medium, the adhesive system caused intense cytotoxicity to the macrophages. The cytotoxic effects were not statistically different (p<0.05) among the experimental groups. In conclusion, chemical components released from SB in aqueous environment were highly toxic to cell culture and thus an inflammatory pulpal response should be considered during the clinical application of dental adhesives.

O objetivo deste estudo foi avaliar o potencial de citotoxicidade do sistema adesivo Adper Single Bond 2 (SB), em função dos tempos pós-polimerização e imersão no meio de cultura para preparação dos extratos, observando-se os níveis de liberação de óxido nítrico (NO) e taxa de sobrevivência celular (MTT assay). Macrófagos alveolares de ratos Wistar foram expostos a 200 μL de extratos obtidos a partir da imersão de amostras do adesivo em meio de cultura (RPMI), imediatamente ou 24 h após sua polimerização, onde permaneceram durante 24 ou 72 h. RPMI puro e lipopolissacarídeos de E. coli foram utilizados como controles negativo e positivo, respectivamente. As células foram levadas à incubadora umidificada por 24 h. Os resultados foram submetidos ao teste "t" de Student (α=5 por cento). As quantidades de NO produzido e células sobreviventes foram significativamente diferentes entre os grupos experimentais e grupos controle, mostrando que, independente do tempo pós-polimerização e tempo de elaboração dos extratos, o sistema adesivo causou uma intensa citotoxicidade sobre os macrófagos. Os efeitos citotóxicos não foram estatisticamente diferentes entre os grupos experimentais. Componentes químicos do SB liberados em meio aquoso podem ser altamente citotóxicos para as células em cultura e, portanto, uma resposta inflamatória pulpar deve ser considerada durante a aplicação clínica de adesivos dentinários.

Simvastatin inhibits induction of matrix metalloproteinase-9 in rat alveolar macrophages exposed to cigarette smoke extract

Matrix metalloproteinase-9 (MMP-9) may play an important role in emphysematous change in chronic obstructive pulmonary disease (COPD), one of the leading causes of mortality and morbidity worldwide. We previously reported that simvastatin, an inhibitor of HMG-CoA reductase, attenuates emphysematous change and MMP-9 induction in the lungs of rats exposed to cigarette smoke. However, it remained uncertain how cigarette smoke induced MMP-9 and how simvastatin inhibited cigarette smoke-induced MMP-9 expression in alveolar macrophages (AMs), a major source of MMP-9 in the lungs of COPD patients. Presently, we examined the related signaling for MMP-9 induction and the inhibitory mechanism of simvastatin on MMP-9 induction in AMs exposed to cigarette smoke extract (CSE). In isolated rat AMs, CSE induced MMP-9 expression and phosphorylation of ERK and Akt. A chemical inhibitor of MEK1/2 or PI3K reduced phosphorylation of ERK or Akt, respectively, and also inhibited CSE-mediated MMP-9 induction. Simvastatin reduced CSE-mediated MMP-9 induction, and simvastatin-mediated inhibition was reversed by farnesyl pyrophosphate (FPP) or geranylgeranyl pyrophosphate (GGPP). Similar to simvastatin, inhibition of FPP transferase or GGPP transferase suppressed CSE-mediated MMP-9 induction. Simvastatin attenuated CSE-mediated activation of RAS and phosphorylation of ERK, Akt, p65, IkappaB, and nuclear AP-1 or NF-kappaB activity. Taken together, these results suggest that simvastatin may inhibit CSE-mediated MMP-9 induction, primarily by blocking prenylation of RAS in the signaling pathways, in which Raf-MEK-ERK, PI3K/Akt, AP-1, and IkappaB-NF-kappaB are involved.

Differential activity of protein kinase C in alveolar and peritoneal macrophages.

The characteristics of protein kinase C activity present in guinea pig alveolar and peritoneal macrophages have been compared and examined. The activity is predominantly cytosolic with preference for phosphatidyl serine as cofactor over other phospholipids. K(m) of protein kinase C for ATP is 30.30 and 54.05 microM in alveolar and peritoneal macrophages respectively. Scatchard plot analysis shows heterogenous binding sites for [3H]PDBu in alveolar macrophages in contrast to peritoneal macrophages showing homogeneous type of binding sites. PMA activates protein kinase C in a dose-dependent manner and shows downregulation at higher concentration in both alveolar and peritoneal macrophages. Endogenous proteins of molecular masses 77, 47, 37 and 16.5 kDa in alveolar macrophages and 77, 47, 38 and 15.5 kDa in pertioneal macrophages are phosphorylated by PKC. These findings suggest that alveolar and peritoneal macrophages possess two different types of protein kinase C activities but phosphorylate similar proteins and exhibit functional similarities in these cells.