How to minimise the effect of tumour cell content in detection of aberrant genetic markers in neuroblastoma

Background:Clinical heterogeneity reflects the complexity of genetic events associated with neuroblastoma (NB). To identify the status of all described genetic loci with possible prognostic interest, high-throughput approaches have been used, but only with tumour cell content >60%. In some tumours, necrotic, haemorrhagic and/or calcification areas influence the low amount of neuroblasts. We evaluated the effect of tumour cell content in the detection of relevant aberrant genetic markers (AGM) diagnosed by fluorescence in situ hybridisation (FISH) on tissue microarrays (TMA) in NB.Methods:Two hundred and thirty-three MYCN non-amplified primary NB included in 12 TMAs were analysed.Results:Presence of AGM reduced event-free survival (EFS) (P=0.004) as well as overall survival (OS) (P=0.004) of patients in the whole cohort. There were no differences in prognostic impact of presence of AGM according to tumour cell content.Conclusion:We propose the use of FISH to diagnose AGM of all NB samples having the above-mentioned areas to determine patient risk

Segmental chromosomal alterations have prognostic impact in neuroblastoma: a report from the INRG project

Background: In the INRG dataset, the hypothesis that any segmental chromosomal alteration might be of prognostic impact in neuroblastoma without MYCN amplification (MNA) was tested. Methods: The presence of any segmental chromosomal alteration (chromosome 1p deletion, 11q deletion and/or chromosome 17q gain) defined a segmental genomic profile. Only tumours with a confirmed unaltered status for all three chromosome arms were considered as having no segmental chromosomal alterations. Results: Among the 8800 patients in the INRG database, a genomic type could be attributed for 505 patients without MNA: 397 cases had a segmental genomic type, whereas 108 cases had an absence of any segmental alteration. A segmental genomic type was more frequent in patients >18 months and in stage 4 disease (P<0.0001). In univariate analysis, 11q deletion, 17q gain and a segmental genomic type were associated with a poorer event-free survival (EFS) (P<0.0001, P=0.0002 and P<0.0001, respectively). In multivariate analysis modelling EFS, the parameters age, stage and a segmental genomic type were retained in the model, whereas the individual genetic markers were not (P<0.0001 and RR=2.56; P=0.0002 and RR=1.8; P=0.01 and RR=1.7, respectively). Conclusion: A segmental genomic profile, rather than the single genetic markers, adds prognostic information to the clinical markers age and stage in neuroblastoma patients without MNA, underlining the importance of pangenomic studies

The C. Elegans Heterochronic Gene lin-46 Affects Developmental Timing at Two Larval Stages and Encodes a Relative of the Scaffolding Protein Gephyrin

The succession of developmental events in the C. elegans larva is governed by the heterochronic genes. When mutated, these genes cause either precocious or retarded developmental phenotypes, in which stage-specific patterns of cell division and differentiation are either skipped or reiterated, respectively. We identified a new heterochronic gene, lin-46, from mutations that suppress the precocious phenotypes caused by mutations in the heterochronic genes lin-14 and lin-28. lin-46 mutants on their own display retarded phenotypes in which cell division patterns are reiterated and differentiation is prevented in certain cell lineages. Our analysis indicates that lin-46 acts at a step immediately downstream of lin-28, affecting both the regulation of the heterochronic gene pathway and execution of stage-specific developmental events at two stages: the third larval stage and adult. We also show that lin-46 is required prior to the third stage for normal adult cell fates, suggesting that it acts once to control fates at both stages, and that it affects adult fates through the let-7 branch of the heterochronic pathway. Interestingly, lin-46 encodes a protein homologous to MoeA of bacteria and the C-terminal domain of mammalian gephyrin, a multifunctional scaffolding protein. Our findings suggest that the LIN-46 protein acts as a scaffold for a multiprotein assembly that controls developmental timing, and expand the known roles of gephyrin-related proteins to development

Detection and treatment of HIV and hepatitis virus infections in Swiss correctional facilities

OBJECTIVES: The aim of the study was to obtain an overview on diagnostic and therapeutic activities concerning hepatitis A, B, C virus and HIV in Swiss prisons.METHODS: A standardized questionnaire was sent to 91 prisons in the German and Italian speaking parts in October 2004; 41 institutions (45%) answered the questionnaire.RESULTS: In almost all prisons serological examinations were not done routinely, but were provided when demanded by inmates or recommended by the medical service. Vaccination against hepatitis A or B infection and initiation of antiviral therapy was possible in most institutions.CONCLUSIONS: Most of the prisons investigated offered diagnostic and antiviral treatment for hepatitis virus and HIV infections. A reported problem was the discontinuation of ongoing treatments or vaccination cycles after discharge. In some cases deficient funding was an obstacle

International consensus for neuroblastoma molecular diagnostics: report from the International Neuroblastoma Risk Group (INRG) Biology Committee

Neuroblastoma serves as a paradigm for utilising tumour genomic data for determining patient prognosis and treatment allocation. However, before the establishment of the International Neuroblastoma Risk Group (INRG) Task Force in 2004, international consensus on markers, methodology, and data interpretation did not exist, compromising the reliability of decisive genetic markers and inhibiting translational research efforts. The objectives of the INRG Biology Committee were to identify highly prognostic genetic aberrations to be included in the new INRG risk classification schema and to develop precise definitions, decisive biomarkers, and technique standardisation. The review of the INRG database (n=8800 patients) by the INRG Task Force finally enabled the identification of the most significant neuroblastoma biomarkers. In addition, the Biology Committee compared the standard operating procedures of different cooperative groups to arrive at international consensus for methodology, nomenclature, and future directions. Consensus was reached to include MYCN status, 11q23 allelic status, and ploidy in the INRG classification system on the basis of an evidence-based review of the INRG database. Standardised operating procedures for analysing these genetic factors were adopted, and criteria for proper nomenclature were developed. Neuroblastoma treatment planning is highly dependant on tumour cell genomic features, and it is likely that a comprehensive panel of DNA-based biomarkers will be used in future risk assignment algorithms applying genome-wide techniques. Consensus on methodology and interpretation is essential for uniform INRG classification and will greatly facilitate international and cooperative clinical and translational research studies

MicroRNA-155 is induced during the macrophage inflammatory response

The mammalian inflammatory response to infection involves the induction of several hundred genes, a process that must be carefully regulated to achieve pathogen clearance and prevent the consequences of unregulated expression, such as cancer. Recently, microRNAs (miRNAs) have emerged as a class of gene expression regulators that has also been linked to cancer. However, the relationship between inflammation, innate immunity, and miRNA expression is just beginning to be explored. In the present study, we use microarray technology to identify miRNAs induced in primary murine macrophages after exposure to polyriboinosinic:polyribocytidylic acid or the cytokine IFN-{beta}. miR-155 was the only miRNA of those tested that was substantially up-regulated by both stimuli. It also was induced by several Toll-like receptor ligands through myeloid differentiation factor 88- or TRIF-dependent pathways, whereas up-regulation by IFNs was shown to involve TNF-{alpha} autocrine signaling. Pharmacological inhibition of the kinase JNK blocked induction of miR-155 in response to either polyriboinosinic:polyribocytidylic acid or TNF-{alpha}, suggesting that miR-155-inducing signals use the JNK pathway. Together, these findings characterize miR-155 as a common target of a broad range of inflammatory mediators. Importantly, because miR-155 is known to function as an oncogene, these observations identify a potential link between inflammation and cancer

Aplicación de parámetros farmacocinéticos/farmacodinámicos al empleo terapéutico de amoxicilina en gatos domésticos

La amoxicilina (AMX) es un antibiótico betalactámico que incluye en su espectro microorganismos grampositivos (estafilococos y estreptococos) y gramnegativos (algunas enterobacterias). La AMX puede administrarse por vía parenteral como formulaciones solubles de liberación y acción rápida o, como formulaciones de liberación sostenida. Los intervalos posológicos habitualmente recomendados son cada 12 h para las formulaciones solubles y, cada 48 h para las formulaciones de depósito. La eficacia clínica de la AMX es "tiempo dependiente" y el indicador empleado para predecir el éxito terapéutico es el T>CIM (tiempo que las concentraciones plasmáticas se encuentran por encima de la CIM del patógeno a tratar). El éxito terapéutico se obtendría con un T CIM ≥40% del intervalo posológico. El propósito de este trabajo es determinar el T CIM para la AMX luego de su administración intravenosa (IV), intramuscular en forma soluble (IMs) e, intramuscular como suspensión de depósito (IMd) a gatos domésticos.Facultad de Ciencias Veterinaria

Aplicación de parámetros farmacocinéticos/farmacodinámicos al empleo terapéutico de amoxicilina en gatos domésticos

La amoxicilina (AMX) es un antibiótico betalactámico que incluye en su espectro microorganismos grampositivos (estafilococos y estreptococos) y gramnegativos (algunas enterobacterias). La AMX puede administrarse por vía parenteral como formulaciones solubles de liberación y acción rápida o, como formulaciones de liberación sostenida. Los intervalos posológicos habitualmente recomendados son cada 12 h para las formulaciones solubles y, cada 48 h para las formulaciones de depósito. La eficacia clínica de la AMX es "tiempo dependiente" y el indicador empleado para predecir el éxito terapéutico es el T>CIM (tiempo que las concentraciones plasmáticas se encuentran por encima de la CIM del patógeno a tratar). El éxito terapéutico se obtendría con un T CIM ≥40% del intervalo posológico. El propósito de este trabajo es determinar el T CIM para la AMX luego de su administración intravenosa (IV), intramuscular en forma soluble (IMs) e, intramuscular como suspensión de depósito (IMd) a gatos domésticos.Facultad de Ciencias Veterinaria

Aplicación de parámetros farmacocinéticos/farmacodinámicos al empleo terapéutico de amoxicilina en gatos domésticos

La amoxicilina (AMX) es un antibiótico betalactámico que incluye en su espectro microorganismos grampositivos (estafilococos y estreptococos) y gramnegativos (algunas enterobacterias). La AMX puede administrarse por vía parenteral como formulaciones solubles de liberación y acción rápida o, como formulaciones de liberación sostenida. Los intervalos posológicos habitualmente recomendados son cada 12 h para las formulaciones solubles y, cada 48 h para las formulaciones de depósito. La eficacia clínica de la AMX es "tiempo dependiente" y el indicador empleado para predecir el éxito terapéutico es el T>CIM (tiempo que las concentraciones plasmáticas se encuentran por encima de la CIM del patógeno a tratar). El éxito terapéutico se obtendría con un T CIM ≥40% del intervalo posológico. El propósito de este trabajo es determinar el T CIM para la AMX luego de su administración intravenosa (IV), intramuscular en forma soluble (IMs) e, intramuscular como suspensión de depósito (IMd) a gatos domésticos.Facultad de Ciencias Veterinaria

Influence of segmental chromosome abnormalities on survival in children over the age of 12 months with unresectable localised peripheral neuroblastic tumours without MYCN amplification.

BACKGROUND: The prognostic impact of segmental chromosome alterations (SCAs) in children older than 1 year, diagnosed with localised unresectable neuroblastoma (NB) without MYCN amplification enrolled in the European Unresectable Neuroblastoma (EUNB) protocol is still to be clarified, while, for other group of patients, the presence of SCAs is associated with poor prognosis. METHODS: To understand the role of SCAs we performed multilocus/pangenomic analysis of 98 tumour samples from patients enrolled in the EUNB protocol. RESULTS: Age at diagnosis was categorised into two groups using 18 months as the age cutoff. Significant difference in the presence of SCAs was seen in tumours of patients between 12 and 18 months and over 18 months of age at diagnosis, respectively (P=0.04). A significant correlation (P=0.03) was observed between number of SCAs per tumour and age. Event-free (EFS) and overall survival (OS) were calculated in both age groups, according to both the presence and number of SCAs. In older patients, a poorer survival was associated with the presence of SCAs (EFS=46% vs 75%, P=0.023; OS=66.8% vs 100%, P=0.003). Moreover, OS of older patients inversely correlated with number of SCAs (P=0.002). Finally, SCAs provided additional prognostic information beyond histoprognosis, as their presence was associated with poorer OS in patients over 18 months with unfavourable International Neuroblastoma Pathology Classification (INPC) histopathology (P=0.018). CONCLUSIONS: The presence of SCAs is a negative prognostic marker that impairs outcome of patients over the age of 18 months with localised unresectable NB without MYCN amplification, especially when more than one SCA is present. Moreover, in older patients with unfavourable INPC tumour histoprognosis, the presence of SCAs significantly affects OS