Amplification of the PLAG-family genes-PLAGL1 and PLAGL2-is a key feature of the novel tumor type CNS embryonal tumor with PLAGL amplification.

Pediatric central nervous system (CNS) tumors represent the most common cause of cancer-related death in children aged 0-14 years. They differ from their adult counterparts, showing extensive clinical and molecular heterogeneity as well as a challenging histopathological spectrum that often impairs accurate diagnosis. Here, we use DNA methylation-based CNS tumor classification in combination with copy number, RNA-seq, and ChIP-seq analysis to characterize a newly identified CNS tumor type. In addition, we report histology, patient characteristics, and survival data in this tumor type. We describe a biologically distinct pediatric CNS tumor type (n = 31 cases) that is characterized by focal high-level amplification and resultant overexpression of either PLAGL1 or PLAGL2, and an absence of recurrent genetic alterations characteristic of other pediatric CNS tumor types. Both genes act as transcription factors for a regulatory subset of imprinted genes (IGs), components of the Wnt/ß-Catenin pathway, and the potential drug targets RET and CYP2W1, which are also specifically overexpressed in this tumor type. A derived PLAGL-specific gene expression signature indicates dysregulation of imprinting control and differentiation/development. These tumors occurred throughout the neuroaxis including the cerebral hemispheres, cerebellum, and brainstem, and were predominantly composed of primitive embryonal-like cells lacking robust expression of markers of glial or neuronal differentiation (e.g., GFAP, OLIG2, and synaptophysin). Tumors with PLAGL1 amplification were typically diagnosed during adolescence (median age 10.5 years), whereas those with PLAGL2 amplification were diagnosed during early childhood (median age 2 years). The 10-year overall survival was 66% for PLAGL1-amplified tumors, 25% for PLAGL2-amplified tumors, 18% for male patients, and 82% for female patients. In summary, we describe a new type of biologically distinct CNS tumor characterized by PLAGL1/2 amplification that occurs predominantly in infants and toddlers (PLAGL2) or adolescents (PLAGL1) which we consider best classified as a CNS embryonal tumor and which is associated with intermediate survival. The cell of origin and optimal treatment strategies remain to be defined.

Flow cytometric analysis of cerebrospinal fluid improves detection of leukaemic blasts in infants with acute lymphoblastic leukaemia.

Infants with acute lymphoblastic leukaemia (ALL) have a high frequency of central nervous system (CNS) involvement. Flow cytometric analysis of cerebrospinal fluid (CSF) was recently demonstrated to be a sensitive method for detecting CNS involvement in childhood ALL. In the present study, CSF from 14 infants was collected at routine lumbar punctures and analysed by multicolour flow cytometry. At initial diagnosis, leukaemic blasts were detected in CSF by flow cytometry in 11 patients (78·6%) compared to seven patients (50%) by cytospin. Larger studies are needed to determine if CSF flow cytometry has prognostic value in infant ALL.

Enhanced recovery, enrichment and detection of Mycobacterium marinum with the Portable Microbe Enrichment Unit (PMEU).

The use of PMEU significantly accelerated the growth of otherwise slowly growing Mycobacterium sp. Compared to the static reference cultures, M. marinum was detected after 24-48h of cultivation in the PMEU Spectrion(®) equipped with infrared (IR) sensors. Parallel static cultures did not exhibit or indicate mycobacterial growth within these time limits, and essentially no growth was found in them. The PMEU approach could provide a powerful tool for the rapid diagnosis and determination of environmental and clinical isolates of slow-growing species of mycobacteria. This approach also provides a method for improving diagnostics for M. tuberculosis and other pathogenic mycobacteria, including their antibiotic resistant forms, which represents a significant health problem worldwide and in Africa in particular.

[Childhood and adolescent cancer was cured--how to support health in adulthood?]. / Lapsuudessa tai nuoruudessa sairastettu syöpä parani--miten tukea terveyttä aikuisena?

The number of long-term survivors after cancer therapy in childhood and young adulthood is increasing. Accordingly, life-long follow-up of significant health problems related to the given cancer therapy is needed as only one third of the survivors will remain free of any physical or psychosocial late effects. At present, national activity is needed to establish a uniform follow-up clinic service to support education, diagnostics, therapy and rehabilitation of these long-term adverse effects after cancer therapy at young age.

Removal of Bovine Insulin From Cow's Milk Formula and Early Initiation of Beta-Cell Autoimmunity in the FINDIA Pilot Study.

OBJECTIVE To test whether weaning to a bovine insulin-free cow's milk formula (CMF) reduces type 1 diabetes mellitus-associated autoantibodies in children at genetic risk. DESIGN Randomized, double-blind pilot trial (Finnish Dietary Intervention Trial for the Prevention of Type 1 Diabetes [FINDIA]). SETTING Three pediatric hospitals in Finland from May 15, 2002, to November 22, 2005. PARTICIPANTS A total of 1113 infants with HLA-conferred susceptibility to type 1 diabetes were randomly assigned to receive study infant formulas; 908 children provided at least 1 follow-up blood sample (last follow-up, June 2009). INTERVENTION The CMF (n = 389), whey-based hydrolyzed formula (WHF) (n = 350), or whey-based FINDIA formula essentially free of bovine insulin (n = 365) during the first 6 months of life whenever breast milk was not available. MAIN OUTCOME MEASURES Primary outcome was beta-cell autoimmunity monitored at ages 3, 6, and 12 months and then annually until age 3 years. Autoantibodies to insulin, the 65-kDa isoform of glutamic acid decarboxylase, and the tyrosine phosphatase-related IA-2 molecule were screened, and islet cell autoantibodies and autoantibodies to zinc transporter 8 were analyzed in infants whose primary screening test results were positive. RESULTS In the intention-to-treat analysis, 6.3% of children in the CMF group, 4.9% of those in the WHF group, and 2.6% of children in the FINDIA group were positive for at least 1 autoantibody by age 3 years. The odds ratios were 0.75 (95% CI, 0.37-1.54) in the WHF group and 0.39 (0.17-0.91) in the FINDIA group when compared with the CMF group. In the treatment-received analysis, the corresponding odds ratios were 0.81 (95% CI, 0.37-1.76) and 0.23 (0.08-0.69). CONCLUSION In comparison with ordinary CMF, weaning to an insulin-free CMF reduced the cumulative incidence of autoantibodies by age 3 years in children at genetic risk of type 1 diabetes mellitus. TRIAL REGISTRATION clinicaltrials.gov Identifier: NCT01055080.

Enterobacterial microflora in infancy - a case study with enhanced enrichment.

OBJECTIVE: To validate PMEU (Portable Microbe Enrichment Unit) method for monitoring the composition and development of infantile intestinal enterobacterial microflora. METHODS: A case study of a boy with neonatal sepsis is presented. During the first 32 months, he was given 19 systemic antibiotic treatments representing seven different antibiotic classes. Seven fecal samples collected at ages from 3.4 to 31.6 months were studied for enterobacterial strains by a combination of enhanced enrichment culture in the PMEU and plate culture. The identification and phenotypic characterization of the isolates was performed by biochemical tests. RESULTS: 37/51 (73%) of the enterobacterial isolates were detected only after the PMEU enrichment. In most samples Escherichia coli strains were predominating, but also several Enterobacter, Klebsiella, Pantoea and Proteus strains could be isolated. It seemed that the antibiotic medications remarkably delayed the development of the intestinal microflora, because first enterobacterial strains were detected only after 6 months of age. CONCLUSIONS: The enrichment step turned out to essentially improve the characterization and monitoring of the intestinal enterobacterial microbiota of infants. Compared to plate culture the amount of isolates was 2.6-fold by the PMEU enrichment. This study gives an idea on the development and succession of microbial communities in the gastrointestinal tract and on the variation of the strains due to the intestinal environmental factors.

Growth and gaseous emissions of pure and mixed small intestinal bacterial cultures: Effects of bile and vancomycin.

Simultaneous cultivations in anaerobiosis, aerobiosis and with microaerobic gas mixture were used to clarify the bile (oxgall) effects on the pure and mixed cultures of enterobacterial strains in simulations in Portable Microbe Enrichment Unit (PMEU) linked with ChemPro100i((R)) gas detector. The effects of vancomycin were evaluated in aerobic cultures. Growth and metabolic activity of cultures were also followed by measuring sugar consumption, pH alterations, and colony counts on BD CHROMagar Orientation plates. Results showed that the two fermentatively different strains of facultative anaerobes, Escherichia coli E 17 and Klebsiella mobilis ATCC 13048 grew in balance regardless of oxygen level, bile acid concentration or other components of the mixed cultures, Bacillus cereus or Staphylococcus aureus. When the evaporations of the mixed cultures of E. coli, K. mobilis and S. aureus were compared with the emissions of the corresponding pure cultures by ChemPro100i((R)) gas sensing detector, the pure cultures of bile resistant E. coli and K. mobilis produced more gaseous components than the mixed culture indicating that these organisms cooperate and use the substrate more effectively together than separately. A survey of the aseptic bacterial isolations from the bile tract in a big University Hospital, (Salzburg, Austria) during 3 years, showed that these bacterial groups dominated. Only 13.24% of the 287 patient samples were sterile, and around 180 strains of both E. coli and Klebsiella/Enterobacter groups were found amongst 973 isolates from 249 patients (together 35.57%). Enterococcus sp. accounted for 246 isolates being the largest group of strains (24.25% of all the isolates). In anaerobiosis it was shown that Klebsiella neutralized the acids produced in the mixed acid fermentation of the E. coli. The ethanol produced from both groups evaporated in the gas stream of the PMEU culturing step and its formation also removes excess acidity from the cultures. The synergistic behaviour and symbiotic function between E. coli and Klebsiella/Enterobacter strains is suggested.

Enhanced enrichment and detection of thermotolerant Campylobacter species from water using the Portable Microbe Enrichment Unit and real-time PCR.

An enhanced enrichment using the Portable Microbe Enrichment Unit (PMEU) with the microaerobic bubbling of broths was applied for the detection of thermotolerant Campylobacter species from water. This PMEU enrichment was compared with the conventional static enrichment of the international standard ISO 17995:2005. In addition, Campylobacter detection after enrichment using a real-time PCR detection was compared with colony counts. The tests with stressed Campylobacter jejuni cells in drinking water indicated that the PMEU enrichment yielded a significantly higher number of Campylobacter cells in the Bolton broth compared with the conventional static incubation. Application of the real-time PCR technique shortened the Campylobacter detection time. This combination of method modifications can be used for Campylobacter detection from water and adds methodological repertoire for the rapid survey and management of waterborne outbreaks.

Breastfeeding stimulates total and cow's milk-specific salivary IgA in infants.

Breastfeeding may increase the rate of mucosal maturation and IgA production. We sought to determine the effect of breastfeeding vs. formula-feeding on the maturation of oral mucosa by measuring the salivary total antibodies and cow's milk protein-specific IgA. Fifty-eight saliva samples were collected from 39 healthy, full term infants. At the age of 3 months (n = 25) eight infants received only breast milk and seventeen formula (cow's milk based n = 10, hydrolysed n = 7) and breast milk; and at the age of 6 months (n = 33) eleven received breast milk, seventeen formula and breast milk and five were not breastfed any more (cow's milk based n = 14, hydrolysed n = 8). Total IgA, IgG, IgM and protein, and beta-lactoglobulin specific IgA were measured from saliva with enzyme-linked immunoassay (ELISA). The antibody results were proportioned to total protein. No differences in antibody levels between the feeding groups were found at 3 months of age. At 6 months, total IgA, total IgM and beta-lactoglobulin-specific IgA were higher among the breastfed infants compared to those receiving formula as supplement to breast milk or not breastfed any more (breast milk vs. any formula p = 0.029, p = 0.015, p = 0.058; breast milk vs. cow's milk formula p = 0.025, p = 0.044, p = 0.038). To conclude, breastfeeding stimulated the mucosal immune system to produce IgA to saliva, which is a marker for immunological maturation and likely provides protection against environmental antigens.

Fast detection of bacterial growth by using Portable Microbe Enrichment Unit (PMEU) and ChemPro100i((R)) gas sensor.

The combination of the Portable Microbe Enrichment Unit (PMEU) cultivation and ChemPro100i((R)) Chemical Detector helps the fast detection of hospital infections. The first alarm of the microbe metabolism and growth in the enrichment culture was achieved in 2-3h when pure cultures of strains isolated from neonatal patients were tested. For the detection of minor concentrations of the bacteria, Bacillus, Staphylococcus, Klebsiella and Escherichia strains were diluted to 0.5-600cells per ml. Their preliminary detection was achieved within 4-5h. The combination of enrichment culture (PMEU) and detection of bacterial products by ion mobility spectrometer is a rapid and sensitive method for diagnosis of bacterial growth. This finding warrants further studies with clinical samples.

Aerobic and anaerobic growth modes and expression of type 1 fimbriae in Salmonella.

The aim of this study was to clarify the growth rates of facultatively anaerobic Salmonella enterica serovar Enteritidis strain in aerobic and anaerobic conditions and the expression of type 1 fimbriae in relation to the growth phases. The cultivation was carried out in a Portable Microbe Enrichment Unit (PMEU) where in same conditions one can grow the cells in parallel by modifying, e.g. aerobiosis only. The results obtained show that although the anaerobic metabolism is generally believed to be a slower producer of biomass or metabolites, in these circumstances S. enterica serovar Enteritidis strain gave comparable growth rates in anaerobiosis with nitrogenation as in aerobic cultures with constant aeration. Fimbrial antigens were produced in the beginning of logarithmic phase of the growth cycle both in the aerobic and anaerobic conditions. The fimbria remained in the presence of oxygen. This capability is possibly used for the intrusion of oxygen containing tissues of host body by the invading pathogens. In conclusion S. enterica serovar Enteritidis strain suspensions grow equally well in constant nitrogenation and aeration, and fimbria were produced in both conditions, during the early logarithmic phase but they prevailed in the presence of aeration.