Differential expression of miRNAs in rhabdomyosarcoma and malignant rhabdoid tumor.

Alveolar rhabdomyosarcoma (RMA) and malignant rhabdoid tumor (MRT) have a frequent metastatic spread and a poor prognosis. Aberrant miRNA expression is often found in metastatic tumors. The aim of this study was to identify specific miRNA expression patterns in these tumors. We analyzed the expression of miRNAs in RMA and MRT in tissue samples and in the rhabdomyosarcoma (RMS) cell lines (Rh30 and RD). Selected target miRNAs were modulated with mimic or inhibitor oligonucleotides. Functional analysis was monitored by flow cytometry and migration assays. A set of 107 differentially expressed miRNAs showed tissue-specific clustering of RMA and MRT. Comparison with the Sarcoma microRNA Expression Database revealed RMA- and MRT-specific miRNAs. Metastatic invasion associated miRNA miR-9 was overexpressed in RMA. miR-200c-inhibiting migration-was lower expressed in RMA than in MRT. Transient transfection of RMS cells with a miR-200c mimic and miR-9( inhibitor did neither increase the expression of the known target E-cadherin nor decrease migration. Expression of E-cadherin could be induced in RD cells using decitabine, but demethylation did not influence cell migration. Despite a comparable high rate of metastatic invasion pediatric RMA and MRT show a different pattern of miRNA expression possibly allowing risk stratification.

PET/MR imaging and optical imaging of metastatic rhabdomyosarcoma in mice.

UNLABELLED: The combination of PET and MR imaging synergizes molecular and morphologic information, allowing better diagnosis in cancer patients. The diagnosis of tumor recurrence in rhabdomyosarcoma is extremely challenging and could be improved with PET/MR imaging. The aim of this study was to validate PET/MR imaging in a disseminated rhabdomyosarcoma mouse model. METHODS: One million alveolar (Rh30) and embryonal (RD) rhabdomyosarcoma cells with stably transfected mCherry and Gaussia luciferase were injected intraperitoneally into NOD/LtSz-scid-IL2Rγnull mice. Nine animals were treated with vincristine (0.75 µg/g/d). Tumor growth was monitored on the basis of serum luciferase activity, optical imaging (OI) of the fluorescent protein mCherry, and sequential PET/MR imaging with 3'-deoxy-3'-(18)F-fluorothymidine ((18)F-FLT) and (18)F-FDG. Immunohistochemical Ki-67 and glucose transporter 1 analysis was used to evaluate tumor cell density and proliferative and metabolic activity. RESULTS: The injection of rhabdomyosarcoma cells led to intraperitoneal tumor growth in 34 of 37 mice (Rh30) and 4 of 9 animals (RD). OI revealed inconsistent results for tumors located near the liver. The detection of tumors in the peritoneal cavity was exclusively possible with sequential PET/MR imaging. PET studies with (18)F-FLT MR imaging were more reliable than (18)F-FDG comparing the tracer uptake and correlation with tumor weight. Treatment with vincristine led to reduced tumor growth, which was efficiently detected with (18)F-FDG PET and MR imaging. Total tumor burden as estimated by PET/MR imaging correlated with the serum luciferase activity. CONCLUSION: We established a unique model of metastatic rhabdomyosarcoma with a high frequency of tumor occurrence and easy monitoring of the tumor growth based on reporter gene expression. The accurate detection of rhabdomyosarcoma requires high soft-tissue contrast provided by the MR imaging and high tracer uptake for PET, which was achieved with (18)F-FLT as the tracer before and (18)F-FDG after treatment with vincristine. PET/MR imaging allows improved diagnosis of experimental rhabdomyosarcoma and therefore might influence clinical therapeutic decisions in the future.

Susceptibility of rhabdomyosarcoma cells to macrophage-mediated cytotoxicity.

The prognosis of advanced stage rhabdomyosarcoma (RMS) is still sobering. In recent years, outcome has not been further improved by conventional therapy. Therefore, novel treatment options such as macrophage-directed immunotherapy have to be investigated. The aim of this study was to analyze the phagocytosis of RMS cells by macrophages and to modulate the susceptibility using monoclonal antibodies and cytotoxic drugs.   Expression of the macrophage activating ligand calreticulin and CD47, the counterpart of the inhibitory receptor SIRPα, was analyzed with Affymetrix mRNA expression arrays and immunohistochemistry on 11 primary RMS samples. Results were verified in two RMS cell lines using flow cytometry and immunocytochemistry. Macrophage cytotoxic activity was quantified by a MTT colorimetric assay in co-culture experiments of RMS cells with monocyte-derived, GM-CSF stimulated macrophages. Gene expression analysis and immunohistochemistry revealed a high expression of CD47 and calreticulin in alveolar and embryonal RMS tissue specimens. Extracellular expression of CD47 on RMS cell lines was confirmed by flow cytometry, whereas calreticulin was exclusively detected in the endoplasmatic reticulum. After co-culturing of RMS cells with macrophages, viability dropped to 50-60%. Macrophage-mediated cytotoxicity was not influenced by a blocking antibody against CD47. However, susceptibility was significantly enhanced after pre-treatment of RMS cells with the anthracycline drug doxorubicin. Furthermore, translocation of calreticulin onto the cell surface was detected by flow cytometry. The immunologic effect of doxorubicin may improve the efficacy of adoptive cellular immunotherapy and chemotherapy of childhood RMS.

Noninvasive ventilation for respiratory distress syndrome: a randomized controlled trial.

CONTEXT: Strategies for reducing exposure to endotracheal ventilation through the use of early noninvasive ventilation has proven to be safe and effective, but the option with the greatest benefits needs to be determined. OBJECTIVE: To determine, in infants with respiratory distress syndrome, if early nasal intermittent positive-pressure ventilation (NIPPV) compared with nasal continuous positive airway pressure (NCPAP) decreases the need for mechanical ventilation. PATIENTS AND METHODS: In this single-center, randomized controlled trial, infants (gestational ages 26 to 33/7 weeks) with respiratory distress syndrome were randomly assigned to receive early NIPPV or NCPAP. Surfactant was administered as rescue therapy. The primary outcome was the need for mechanical ventilation within the first 72 hours of life. RESULTS: A total of 200 infants, 100 in each arm, were randomly assigned. Rates of the primary outcome did not differ significantly between the NIPPV (25%) and NCPAP (34%) groups (relative risk [RR]: 0.71 [95% confidence interval (CI): 0.48­1.14]). In posthoc analysis, from 24 to 72 hours of life, significantly more infants in the NIPPV group remained extubated compared with those in the NCPAP groups (10 vs 22%; RR: 0.45 [95% CI: 0.22­0.91]). This difference was also noted in the group of infants who received surfactant therapy, NIPPV (10.9%), and NCPAP (27.1%) (RR: 0.40 [95% CI: 0.18­0.86]). CONCLUSIONS: Early NIPPV did not decrease the need for mechanical ventilation compared with NCPAP, overall, in the first 72 hours of life. However, further studies to assess the potential benefits of noninvasive ventilation are warranted, especially for the most vulnerable or preterm infants.

Cetuximab promotes immunotoxicity against rhabdomyosarcoma in vitro.

Multidrug resistance is a common problem in the treatment of childhood rhabdomyosarcoma (RMS). Therefore, novel treatment regimes such as immunotherapy have to be evaluated. The aim of this study was to detect possible targets on RMS cells and to investigate whether corresponding humanized antibodies could be used to treat RMS. Screening for potential targets common for different subtypes of RMS was carried out with Affymetrix mRNA expression arrays on 12 primary RMS samples. Subsequent pathway analysis revealed the epidermal growth factor receptor (EGFR) as a potential target for antibody therapy. Expression of EGFR and binding of its specific antibody Cetuximab to embryonal RMS cell lines RD and A-204 and alveolar RMS Rh30 were monitored by flow cytometry. Cetuximab activity was quantified by proliferation assay on RMS cells, and by antibody dependent cellular cytotoxicity assay with peripheral blood mononuclear cells (PBMCs). Gene expression analysis revealed a high expression of EGFR in all embryonal RMS compared with alveolar RMS. The EGFR specific antibody, Cetuximab binds to Rh30 and to RD but not to A-204 cells. Proliferation of these cells was influenced neither by Cetuximab nor by the growth factor EGF. However, cell dependent cytotoxicity of PBMCs to RMS cells such as Rh30 and RD was enhanced specifically by Cetuximab. These promising Cetuximab effects justify analysis under in vivo conditions using suitable models.

Fatores de risco para o desenvolvimento de sepse neonatal tardia em uma unidade de terapia intensiva / Risk factors for the development of tlate-oset neonatal sepsis in an intensive care unit

Objetivo: identificar os fatores de risco maternos e neonatais para o desenvolvimento da sepse neonatal tardia. Casuística e métodos: foi realizado um estudo transversal, que avaliou um grupo de 100 rcém-nascidosno período de outubro de 2006 a junho de 2007 internados na UTI neonatanal do Hospital Universitário da Universidade Federalde Alagoas...

Objective: to Identify mother and neonatal risk factors for the development of late-onset neonatal sepsis. Case study and methods: a transversal study was carried and from Ovtober 2006 to June 2007,evaluating a group of 100 newborns admitted at the Neonatal Intensive Care Unit od Alagoas Federal University Hospital...

Perfil epidemiológico das mães e dos recém-nascidos que foram a óbito no Instituto Materno-Infantil de Pernambuco (IMIP), no período 1991-1992 / The mothers'epidemic profile and of the newly born that were to death in the Maternal Infantile Institute of Pernambuco (IMIP), in the period 1991-1992

Estudo descritivo em 257 recém-nascidos que foram a óbito no Serviço de Neonatologia da Maternidade do Instituto Materno Infantil de Pernambuco (IMIP), no período de 1991 e 1992, a fim de descrever e relacionar o perfil das principais características das mães e dos recém-nascidos. As características maternas analisadas foram: procedência, idade, grau de instrução, assistência pré-natal, tipo de parto e presença de doenças perinatais. Nos recém-nascidos observou-se o peso, idade gestacional, classificação idade-peso, sexo, Apgar no 1§ e 5§, minuto e idade do óbito. Apesar de a maioria das gestantes terem acesso ao pré-natal, apenas 27,1 por cento realizaram mais de quatro consultas e em 56,2 por cento dos casos, uma ou mais doenças estavam presentes. Não houve significância estatística entre idade materna e peso ao nascer e entre assistência pré-natal e frequência de baixo peso, porém nas gestantes que fizeram menos de quatro consultas no pré-natal houve maior incidência desse grupo. Os resultados mostram que é fundamental que se controle o maior número possível de fatores associados ao nascimento de crianças de baixo peso, para que se consiga uma redução da mortalidade neonatal

Marcadores inflamatórios (interleucina-1beta, interleucina-6, interleucina-8, interleucina-10 e proteína C-reativa) em recém-nascidos pré-termo com sepse neonatal precoce e corioamnionite histológica / Inflammatory markers (interleukin-1b, interleukin-6, interleukin-8, interleukin-10 and C- reactive protein) in preterm newborn with early-onset sepsis and histologic chorioamnionitis

Foram estudados 102 recém-nascidos pré-termo (RN), procedentes da maternidade Escola Santa Mônica, em Alagoas, no período de agosto de 2001 a agosto de 2002, cujo objetivo foi analisar o comportamento dos marcadores inflamatórios (IL-1b; IL-6; IL-8; IL-10 e PCR) no RN pré-termo e sua associação com sepse precoce, corioamnionite histológica e mortalidade. Constatou-se na sepse precoce um aumento significativo (p < 0,05) da IL-1 (1o dia de vida); IL-6, IL-8 (3o dia de vida); e IL10 e PCR (1o e 3o dias de vida). Nas placentas com corioamnionite os valores das IL-1b e PCR no 1o dia de vida foram significativos (p < 0,05). Não houve diferença na mortalidade, concluindo-se que a utilização desses marcadores foi eficaz para o diagnóstico da sepse neonatal precoce e de corioamnionite.From August 2001 to August 2002, 102 preterm newborn from Santa Mônica Hospital, in the State of Alagoas, Brazil, were studied. The objective was the analysis of inflammatory markers (IL-1b; IL-6; IL-8; IL-10 and CRP) effects in preterm infant and their association with early-onset sepsis, histologic chorioamnionitis and mortality. It was detected in early-onset sepsis a significant increase (p < 0,05) of IL-1 (1st day of life); IL-6, IL-8 (3rd day of life); and IL-10 and CRP (1st and 3rd day of life). In placents with chorioamnionitis, the values of IL-1b and CRP at the 1st day of life were significative (p < 0,05). There were no difference between the values of these markers and mortality. The conclusion is that the use of these markers was efficient to preterm neonatal sepsis and chorioamnionitis diagnosis...