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Introduction NHS England announced a new nurse angiographer role with the purpose of reducing hospital waiting lists for elective angiography. Methods Development of a standardised operating procedure for training in Radial Arterial Access (RAA), guidance on how to minimise the risk and recognise complications and post procedural limb management. Obtaining approval at the new procedures committee. Training program 50 consultant supervised nurse led procedures for (RAA), establishing expertise in 100 further procedures, followed by roll out of the program to other nursing staff. Results During training (RAA) cannulation was successful on 1st attempt in 42 out of 50 (success rate allowing for two attempts 84%), with no complications recorded. Successful access by a more experienced operator N = 7, failed (RAA) N = 1. The following 100 were performed independently on unselected patients (success rate 81%; (1st pass 70)), with no complications, need for alternate access routes, N = 1. The following table details the success rate and patient comfort score recorded during 104 procedures performed by a nurse specialist and trainee registrars. Conclusion Nurse-led (RAA) is safe and feasible and has the potential to improve catheter laboratory efficiency, patient satisfaction, standardise procedure training and improve staff retention. Our next steps will be to train more nurses in radial access and progress to nurse led angiography. Conflict of Interest None

Background Despite major treatment attempts, the prognosis for pediatric diffuse intrinsic pontine gliomas (DIPGs) remains dismal. Gliomas are highly vascularized tumors, suggesting that the prevention of vessel formation by anti-angiogenic treatment might be effective. Procedure Forty-one pediatric patients with DIPG were treated according to the Angiocomb protocol, starting with radiotherapy combined with topotecan and followed by anti-angiogenic triple medication consisting of thalidomide, etoposide, and celecoxib. Overall survival, radiological response, quality of life, requirement of corticosteroids, and adverse effects were monitored. Eight patients treated with only radiotherapy were used as controls. Results For study patients, the 12 and 24 months overall survival was 61% and 17%, respectively. The median overall survival was 12 months (range 4–60 months). Four radiological complete responses were seen, of which two were transient. Radiologically, 56% of the tumors reduced in size and 78% in signal intensity. Study patients were able to visit school or daycare and walk for a significantly longer time compared to controls (Log Rank 0.036 and 0.008, respectively). Adverse effects were generally minor. Conclusions The Angiocomb protocol created a noticeable share of long-term survivors and was well tolerated, suggesting that anti-angiogenic therapy for patients with DIPG should be studied more in the future. Pediatr Blood Cancer 2014;61:1603–1609. © 2014 Wiley Periodicals, Inc.

Cel Celem pracy byla ocena wybranych czynnikow ryzyka mających wplyw na przebieg procesu dojrzewania u dzieci po alogenicznym przeszczepie krwiotworczych komorek macierzystych (allo-HSCT). Material i metody Prospektywnym badaniom poddano grupe 21 pacjentow (13 dziewczynek, 8 chlopcow) w wieku 10–20 lat (mediana 14). Dzieci leczone byly z powodu: ostrej bialaczki limfoblastycznej (ALL) – 5 pacjentow, ostrej bialaczki nielimfoblastycznej (ANNL) – 4 pacjentow, przewleklej bialaczki szpikowej (CML) – 5 pacjentow, zespolu mielodysplastycznego (MDS) – 2 pacjentow, chloniaka nieziarniczego (NHL) – 1 pacjent, mlodzienczej bialaczki mielomonocytowej (JMML) – 1 pacjent, ciezkiej postaci anemii plastycznej (SAA) – 1 pacjent, niedokrwistości Blackfana-Diamonda – 1 pacjent oraz miesaka prązkowanokomorkowego (RMS) – 1 pacjent. Typ zastosowanego przeszczepu: HLA ID SIB (11), HLA MM REL (3), MUD-HSCT (7). Najcześciej stosowano wysokodawkowaną chemioterapie w postaci: busulfan/melfalan (BU/MEL-2); busulfan/cyklofosfamid/etopozyd(BU/CY/VP-5); busulfan/cyklofosfamid/ATG (BU/CY/ATG 4); 3 dzieci poddano napromienianiu calego ciala (total body irradiation–TBI). Napromienianie OUN przeprowadzono u 5 pacjentow: 18 Gy (4), 24 Gy (1). U 7 dzieci zastosowano kortykoterapie >28 dni przed przeszczepem i u 10 po przeszczepie. Analizie poddano wyniki nastepujących badan: stezenia gonadotropin – hormon folikulotropowy (FSH), hormon luteinotropowy (LH); prolaktyny (PRL); estradiolu, testosteronu, wolnej 3,5,3’-trijodotyronine (fT3), wolną tyroksyne (fT4), przeciwciala przeciw peroksydazie tarczycowej (ATPO), testy: z tyreoliberyną (TRH) i z gonadoliberyną (GnRH). Wyniki U 12 dzieci (9 dziewczynek, 3 chlopcow) stwierdzono nieprawidlową funkcje gonad od 10 do 72 miesiecy (średnio 28 miesiecy po przeszczepie): FSH 17,9–170 mIU/ml (średnio 39,89 mIU/ml), LH 6–40,1 mIU/ml (średnio 12,78 mIU/ml), estradiol Wnioski Nieprawidlowa funkcja gonad jest czestym nastepstwem stosowania chemioterapii wysokodawkowanej: cyklofosfamidu i busulfanu. Nasze badania nie wykazaly wplywu melfalanu i TBI na wystąpienie zaburzen dojrzewania plciowego, w odroznieniu od doniesien innych autorow.

Despite major treatment attempts, the prognosis for pediatric diffuse intrinsic pontine gliomas (DIPGs) remains dismal. Gliomas are highly vascularized tumors, suggesting that the prevention of vessel formation by anti-angiogenic treatment might be effective.Forty-one pediatric patients with DIPG were treated according to the Angiocomb protocol, starting with radiotherapy combined with topotecan and followed by anti-angiogenic triple medication consisting of thalidomide, etoposide, and celecoxib. Overall survival, radiological response, quality of life, requirement of corticosteroids, and adverse effects were monitored. Eight patients treated with only radiotherapy were used as controls.For study patients, the 12 and 24 months overall survival was 61% and 17%, respectively. The median overall survival was 12 months (range 4-60 months). Four radiological complete responses were seen, of which two were transient. Radiologically, 56% of the tumors reduced in size and 78% in signal intensity. Study patients were able to visit school or daycare and walk for a significantly longer time compared to controls (Log Rank 0.036 and 0.008, respectively). Adverse effects were generally minor.The Angiocomb protocol created a noticeable share of long-term survivors and was well tolerated, suggesting that anti-angiogenic therapy for patients with DIPG should be studied more in the future.

Background The presence of metabolically important genetic polymorphisms may affect treatment efficacy in patients with malignancies. The objective of this prospective multicenter study was to evaluate the role of selected polymorphisms of genes associated with metabolism of chemotherapeutic drugs as prognostic markers in children with acute lymphoblastic leukemia. Procedure Genotyping for the presence of 7 genetic variants in 403 patients and analysis of death cases were performed. Results Thirty-one children died before reaching remission maintenance phase. Genetic analysis revealed in this group increased frequency of homozygosity for c.677C>T polymorphism of the MTHFR gene (26% vs. 8% in the survivors; OR 4.09; 95% CI 1.67–10; adjusted for multiple testing P = 0.028). Conclusion Our data suggest that modification of anti-leukemic treatment should be considered in patients homozygous for c.677C>T polymorphism. Pediatr Blood Cancer 2009;52:364–368. © 2008 Wiley-Liss, Inc.

The aim was to determine whether outcome of unrelated donor transplantation for severe aplastic anemia has improved in recent years and whether this is due to patient selection or better transplant technology. We analyzed 498 patients transplanted during 1990-2005. By running univariate regression models dichotomizing year of transplantation we defined 1998 as the year of the most significant change in survival. Five-year survival increased from 32+/-8% before 1998 to 57+/-8% after 1998 (P

The kinetics of the CH2I + NO2, CH2Br + NO2, and CHBrCl + NO2 reactions have been studied at temperatures between 220 and 360 K using laser photolysis/photoionization mass spectrometry. Decays of radical concentrations have been monitored in time-resolved measurements to obtain reaction rate coefficients under pseudo-first-order conditions. The bimolecular rate coefficients of all three reactions are independent of the bath gas (He or N2) and pressure within the experimental range (2-6 Torr) and are found to depend on temperature as follows: k(CH2I + NO2) = (2.18 +/- 0.07) x 10(-11) (T / 300 K)(-1.45) (+/- 0.22) cm3 molecule(-1) s(-1) (220-363 K), k(CH2Br + NO2) = (1.76 +/- 0.03) x 10(-11) (T/300 K)(-0.86) (+/- 0.09) cm3 molecule(-1) s(-1) (221-363 K), and k(CHBrCl + NO2) = (8.81 +/- 0.28) x 10(-12) (T/300 K)(-1.55) (+/- 0.34) cm3 molecule(-1) s(-1) (267-363 K), with the uncertainties given as one-standard deviations. Estimated overall uncertainties in the measured bimolecular reaction rate coefficients are about +/-25%. In the CH2I + NO2 and CH2Br + NO2 reactions, the observed product is formaldehyde. For the CHBrCl + NO2 reaction, the product observed is CHClO. In addition, I atom and iodonitromethane (CH2INO2) or iodomethyl nitrite (CH2IONO) formations have been detected for the CH2I + NO2 reaction.

Allogeneic hematopoietic cell transplantation (HCT) in children with myelodysplastic syndrome (MDS) remains a challenge due to the toxic conditioning regimens administered to minimize the risk of relapse in the HLA-matched or of graft rejection in the HLA-mismatched settings. In the absence of matched sibling donors, alternative donors such as unrelated and/or partially matched family sources remain risky, yet the only available, options. Herein we report the results of HCT from alternative donors in 14 children with different subtypes of MDS (juvenile myelomonocytic leukemia [JMML] n = 9; myelodysplastic syndrome [MDS] refractory anemia n = 3; MDS refractory anemia with excess of blasts in transformation n = 2) transplanted at our institution. The median time from diagnosis to HCT was 9 months (range 4 to 90 months). The variety of HCT types included: unrelated peripheral blood progenitor cell transplantation (PBPCT) (n = 2), partially matched family donor T-cell-repleted BMT/PBPCT (n = 6), and haploidentical T-cell-depleted PBPCT (n = 6). Five of 14 patients remain alive at 7 to 37 months posttransplant (including two patients after partially matched family donor BMT, two patients after haploidentical T-cell-depleted-PBPCT, and one after unrelated-PBPCT, respectively). The major complications were: primary graft failure in the haploidentical T-cell-depleted-setting or graft-versus-host disease (GvHD) in T-cell-repleted partially matched family or unrelated settings, respectively. Despite the high transplant-related mortality rate in this series, allogeneic HCT from alternative donors remains an interesting solution for children with MDS who lack matched sibling donors. Due to improved immune reconstitution, despite an increased risk of GvHD, T-cell-repleted transplants from single HLA-mismatched family donors remain a valuable option for children without matched donors. Splenectomy prior to HCT may positively affect the posttransplant course in patients with overt splenomegaly for example those afflicted with JMML.

Abstract 4007 Refractory cytopenia in childhood (RCC) is the most common subtype of childhood MDS. It is characterized by persistent cytopenia with Disclosures: No relevant conflicts of interest to declare.

Abstract 4100 Introduction: Among drug-modifying elements age, sex, coexistent diseases, interactions with other drugs, as well as genetically determined altered drug response are included. Analysis of selected gene polymorphisms may lead to an increase treatment safety and efficacy in developing individual therapies. The objectives of the study: 1. Determination of the frequency of the allele variants: TPMT*2, *3A and *3C in population of Polish children treated for acute leukemia. 2. Analysis of the influence of TPMT genetic polymorphism on the occurrence of therapeutic and/or adverse effects such as hematological disorders, hepatotoxicity and nephrotoxicity after thiopurine administration. 3. Determination of the significance of pharmacogenetic profile for dosage optimizing of 6-mercaptopurine and 6-tioguanine. 4. Determination of the leukemia relapse risk in relation to TPMT polymorphism. Patients and Methods: The study group consisted of 210 patients (121 boys, 89 girls) treated for acute leukemia (167 ALL, 43 AML), with median age 7 (range 1-18) yrs, at following time points: on diagnosis, during intensive chemotherapy and after therapy cessation. Adverse treatment effects during the entire observation period including 6 particular stages was classified according to WHO toxicity scale in each child. Either bone marrow or peripheral blood samples were used for the analyses. DHPLC and RFLP was used for genotype determination for TPMT on selected polymorphisms: TPMT*2,*3A,*3C. DNA fragment amplification was used with relevant starters, followed by HpyF10VI (MwoI) and XmiI (AccI) restricting enzymatic digestion. RFLP marking results were validated by DHPLC. Results: 1. For TPMT IM phenotype (“medium” metabolizer) was detected in 7.65% patients (n=16) and PM phenotype (“slow” metabolizer) in 0.47% cases (n=1), the most frequent polymorphic variant being TPMT*3A (72%). 2. Analysis of changes in selected morphology indicated that in TPMT *2, *3A or *3C polymorphism carriers' group there is a significant decrease in white blood count (p=0.0025), platelet count (p=0.0014) and neutrophil count (p=0.019) in comparison with individuals with the wild variant (TPMT*1/*1). In patients with TPMT allelic variants, longer periods of thrombocytopenia in relation to other hematologic parameters were observed (re-induction period: p=0.041, maintenance therapy: p=0.03). In patients with IM phenotype, significantly lower haemoglobin levels were observed only in the end stage of chemotherapy (p=0.032). In patients with ALL and IM the observed risk of leukopenia, (p=0.006), neutropenia (p=0.015) and thrombocytopenia (p=0.011) as well as hepatotoxicity: GOT (p=0.009), LDH (p=0.0014) during the entire treatment period and particular study points (WBC: re-induction p=0.004; PLT: re-induction p=0.045 and maintenance therapy p=0.043) was significantly more frequent. 4. In TPMT polymorphic variant carriers, the relapse of primary disease was statistically less frequent (p=0.0341F). 5. Death risk was not relevant to TPMT genotype. Conclusions: 1. Frequency of detected TPMT single nucleotide polymorphism in children treated for acute leukemia is consistent with the published data for European population. 2. Thiopurine administration is related to the increase in hematological toxicity (leukopenia, neutropenia and thrombocytopenia) in TPMT heterozygotes. 3. Treatment with mercaptopurine or thioguanine should be regarded as an indication for genotyping. 4. TPMT single nucleotide mutation occurrence in children is related to lower risk of relapse in acute leukemia and might be included in the risk group stratification. Disclosures: No relevant conflicts of interest to declare.

Abstract 1715 Poster Board I-741 Introduction Gene polymorphism coding for drug-metabolizing enzymes may cause individual differences in effectiveness and toxicity of many medications, including cytostatics. Although intensified treatment resulted recently in better prognosis of leukemia, many adverse effects are still observed during therapy resulting from nonspecific activity and narrow therapeutic index of anti-cancer drugs. The objectives of the study: 1. Determining the frequency of selected allele variants: CYP2C9, CYP2C19 in population of Polish children treated for acute leukemia. 2. Analysis of the role of CYP2C9 and CYP2C19 in cyclophosphamide metabolism. 3. Determininig the influence of carrying selected CYP2C9 and C19 polymorphism on cyclophosphamide metabolism in form of increased adverse reaction risk (according to WHO score), relapse and other infringements in treatment protocols and procedures following the use of standard doses of alkylating agents. Patients: 184 patients (106 boys, 78 girls) were examined, age between 1 and 18 (median age 7 years, average age 7.64 years, SD=5.3) treated for acute leukemia (ALL:167, AML:17) at various time points: on diagnosis, during cytostatic treatment and after therapy cessation. All patients were treated with current uniform treatment protocols and procedures. Adverse reactions in each child was recorded according to WHO toxicity scale, during the entire length of observation period and in particular stages (6 periods). Methods Either bone marrow or peripheral blood samples were used for the analyses. DNA isolation procedure was carried out with the use of standard tests. CYP2C9*2,*3 and CYP2C19*2 allelic variant determination was carried out with real-time PCR with double-marked probe hydrolysis with the use of Pre-Developed TaqMan Assay Reagents for Allelic Discrimination sets. PCR products fluorescence was carried out on 7900HT Fast Real-Time PCR device. Results 1. For CYP2C9, IM phenotype was present in 22.3% patients (n=41) and PM phenotype in 7.6% cases (n=14), the most frequently detected polymorphic variant being CYP2C9*2 (75.3%). In case of CYP2C19, IM phenotype occurred in 22.3% patients (n=41) and PM phenotype in 7.1% (n=13). 2. Toxicity risk analysis in patients with acute leukemia did not reveal statistically significant differences depending on CYP2C9 genotype. In patients with CYP2C19*2 allelic variant, statistically lower level of alkaline phosphatase (p=0.013) was observed during the entire treatment and in particular stages (I: p=0.0096; II: p=0.022; III: p=0.038; IV:p=0.0024; V:p=0.044). Lower alkaline phosphatase levels were observed only in patients with ALL (p=0.026). Lower transaminase levels after completion of intensive treatment phase were observed in patients with CYP2C19 mutations (GOT, p=0.044). In patients with CYP2C19 and CYP2C9 polymorphic variants no dependence of risk of adverse reactions with correlation to cyclophosphamide and its dose was observed. 3. In patients with CYP2C9 and CYP2C19 polymorphism no significant relation was discovered. 4. Death risk was not relevant to CYP2C9 or CYP2C19 genotype. Conclusions 1. Occurrence of slow metaboliser phenotype for CYP2C9 and CYP2C19 is more frequent in patients with acute leukemia than in the untreated population. 2. CYP2C9 and CYP2C19 cytochromes are involved in cyclophosphamide metabolism but do not play a major role in drug metabolism. 3. CYP2C9 and CYP2C19 single nucleotide polymorphism is correlated neither with an increased risk of adverse reactions or death during therapy, nor with a relapse of acute leukemia in children. Disclosures No relevant conflicts of interest to declare.

Background: Neoplasms in children are uncommon diseases however they have broad impact on the society. Despite novel therapeutic strategies for children suffering from leukemia or solid tumor, the mortality rate of the disease is still high. The endogenous nitric oxide synthesis inhibitor asymmetrical dimethyl-L-arginine (ADMA) is considered to be a strong cardirovascular risk factor and may lead to new therapeutic strategies. Our previous findings indicate that elevated plasma level of ADMA is correlated with worse prognosis in adult patients with NHL. There are no data available pointing at metabolites of NO-pathway nor clinical evidence as prognostic factor of oncological treatment in children. Aim: To investigate the relation between elevated plasma level of NO-pathway metabolites and well established markers of clinical stage of the disease. Material and method: 12 children with acute leukemias (ALL, AML), 13 with solid tumor (PNET, osteosarcoma, Wilms’ tumor, RMS) during hemato-oncological treatment were recruited to a pilot study and N=15 healthy children to a control group. Plasma concentrations of ADMA, SDMA, L-Arg was determined. Relationship to clinical stage of the disease, biochemical and blood markers was tested. Results Table 1: Differences in mean plasma concentrations of arginine, ADMA and SDMA in pts with leukemia vs pts with solid tumor vs control group Control:N=15 Mean ±SD Median ±SEM Leukemia;N=12 Mean±SD Median ±SEM Solid tumour; N=13 Mean±SD Median ±SEM P L-Arg 68,58±24,29 60,19±6,27 31,28±8,63 30,82±2,49 49,59±10,81 49,18±3,00 0,0000 ADMA 0,62±0,10 0,61±0,02 1,17±1,10 0,53±0,32 0,98±1,10 0,46±0,31 NS SDMA 0,44±0,18 0,39±0,04 1,18±1,10 0,80±0,32 1,56±1,02 1,09±0,28 0,0024 ARG/ADMA 112,8±46,28 107,4±11,94 67,40±54,31 69,48±15,68 228,82±451,44 117,35±125,21 NS Table 2:Significant correlations in group of pts with solid tumor and in patients with leukemia LEUKEMIA; N=12 R p ADMA & Ca 0,78 0,008 SDMA & Ca 0,75 0,011 SDMA & CRP −0,78 0,021 Arg/ADMA & K −0,72 0,019 Arg/ADMA & Ca −0,64 0,043 Arg& MONOCYTE 0,65 0,043 ADMA & TOT.BILIRUB. −0,92 0,000 ADMA & TOT.PROTEIN 0,75 0,032 SDMA & EOSINOPHILE 0,73 0,025 SDMA & TOT.BILIRUB −0,82 0,007 Arg/ADMA & TOT.BILIRUB. 0,81 0,009 SOLID TUMOR; N=15 R p Arg &RBC 0,70 0,016 Arg & Hb 0,64 0,035 Arg & LYMPHOCYTE 0,81 0,003 Arg & EOSINOPHILE 0,76 0,011 ADMA &MONOCYTE −0,78 0,008 ADMA & TOT.PROTEIN 0,62 0,040 ADMA & Hb 0,64 0,035 Arg/ADMA &MONOCYTE 0,82 0,004 Arg/ADMA & BASOPHILE 0,65 0,043 Conclusions:A lower plasma level of L-Arg was found both in pts with leukemia and solid tumors. Higher plasma concentration of SDMA was found in patients with leukemia and solid tumor compared to the control group. Further studies are necessary to clarify the significance of these findings.

Background The increased number of successfully treated pts with HSCT necessitates continuous observation focused on endocrinological complications. Significant risk factors for endocrine dysfunctions are age and nutrition state before HSCT, high dose chemotherapy HDC/T, total body irradiation (TBI), prolonged steroid therapy for GvHD and cranial irradiation (CI) given prior to the transplant maneuver. Objective The aim of this study was to evaluate the occurrence of endocrine complications, in particular disorders of growth and thyroid function, abnormal gonadal function and dysfunction in glucose metabolism in children treated with allo-, and auto-HSCT. Material and methods The investigated group consisted of : - 16 patients after auto-HSCT (7 girls, 9 boys) age at transplant maneuver 3–20 years (average 9,56±) transplanted for AML (5), NHL (3), NBL (3), embryonal cancer (2), medulloblastoma, Ewing sarcoma/PNET, HES. High dose chemotherapy (HDC/T) included: BU/MEL (7), BEAM (3). - 30 patients after allo-HSCT (20 girls, 10 boys) age at transplant manoveur 3–17 years (average 9,56±). Indications were ALL (11), AML (5), CML (6), MDS (2), NHL, JMML, SAA, Blackfan-Diamonds anaemia, SCID, RMS. According to donor availability there were MSD (13) patients, MUD (11) and HLA-mismatched related (6). The preparative regimen consisted of HDC/T usually BU/MEL (3); BU/CY/VP (6); BU/CY/ATG (5), VP/ATG/TBI (3). CI prior to grafting received 19 children: auto-HSCT (6), allo-HSCT (13) and TBI 6 patients. Prolonged high steroid doses (beyond 28 days) received 18 children: auto-HSCT (4) and allo-HSCT (14) before, and 20 after HSCT. Endocrine function was evaluated in average after 24 months in auto-setting and after 26 months in allo-setting. TSH, fT3, fT4, IGF-1, IGFBP-3, HbA1c, FSH, LH, PRL, estradiol and testosterone levels, oral glucose tolerance test, GH test, TRH test, LHRH test (LH, FSH) was performed in each patient. The University Research Committee for Bioethics did consent to this investigation protocol. Results 16 children (11girls and 5 boys) presented with abnormal puberty: 43,75% after auto- and 34,61% after allo-HSCT. Hypothyroidism occurred in 5 patients necessitating thyroid hormone substitution. Hyperthyroidism was not diagnosed. Elevated level of aTPO was noted in one child only. Impaired GH-excretion after stimulation was documented in 14 pts. Growth and maturation stature is still an ongoing observational study in this subgroup. Growth impairment was documented in 8 patients (6 girls, 2 boys) 13 to 70 months after allo- transplantation (average 36 months). Three children from above group received CI. Growth hormone substitution was instituted in 1 girl (ALL, HLA MM REL, CI). (SDS Conclusions Early endocrinological care of children treated with both auto-, as well as allo-HSCT is necessary due to a significant risk for hormonal disorders. Early diagnosis of impaired endocrine function together with early substitution therapy will significantly improve the quality of life after HSCT.

Background Preparative regimen consisting of high dose chemotherapy HDC/T and/or cranial irradiation (CI) in combination with prolonged steroid therapy may cause abnormal gonadal function. Objective Evaluate puberty disorders in children occurring after HSCT as single center experience. Material and methods The study group consisting of 21 patients (13 girls and 8 boys) age 10–20 years at grafting (median 14 years) was prospectively evaluated post grafting. The indications for HSCT were: ALL (5), AML (4), CML (5), MDS (2) and NHL, JMML, SAA, Blackfan Diamond anemia, RMS. According to donor availability 11 children had mached sib donors (MSD), 7 matched unrelated donors (MUD) and 3 patients HLA-mismatched related donors. The preparative regimens consisted of HDC/T usually BU/MEL (2pts); BU/CY/VP (5pts); BU/CY/ATG (4pts) and total body irradiation (TBI) received 3 patients with 12 Gy. CI prior to grafting was delivered to 5 children: 18 Gy (4) and 24 Gy (1). Prolonged high doses steroids (beyond day +28) received 7 children prior to- and 10 children after grafting. Endocrine function after allo-HSCT was evaluated from 10 to 42 (median 26 months). Analysis of LH, FSH, PRL, oestradiol and testosterone levels, fT3, fT4, aTPO, TRH test and LHRH test was performed in all study patients. Bone age according to Pyle-Grulich method was estimated. Results: Abnormal gonadal function was present in 12/21 children (9 girls, 3 boys) in average 28 months after HSCT Primary amenorrhea occurred in 3, secondary amenorrhea in 6 girls. Delayed bone age was documented in 2 children in average 30 mths post grafting. No significant correlation was found between children’s age during allo-HSCT and occurrence of puberty disorders. Gender of children had no effect on evalated variables. Time elapsing from HSCT did not influence the occurrence of abnormal gonadal function. No association was found between abnormal gonadal function and TBI, prolonged high doses steroids prior to- and after HSCT. Occurrence of puberty disorders was statistically significantly blunted by conditioning regimen containing Cyclophosphamide (p=0,019) and Busulfan (p=0,0179) while Melfalan did not turn out such a risk factor. PRL levels and thyroid function do not have any determination on occurrence of abnormal gonadal function after HSCT. Conclusion: Abnormal gonadal function as late effect frequently occure after HSCT. High-dose Cyclophoshamide and Busulphan tretaed children often had blunted puberty. Melfalan and TBI, however, did not cause such an effect reported by others. Endocrinological care for patients after HSCT is necessary in order to: - Discover the impaired endocrine function - Initiate correct hormonal substitution therapy - Significantly improve the quality of life after allo- HSCT and - Secondary prevent treatment related complications.

The aim of the study was to analyze the correlation between the number of transplanted CD34+ cells and CD3+ cells and the incidence of severe Graft versus Host Disease (GvHD) in children undergoing allogeneic hematopoietic cell transplantation from HLA-matched or mismatched unrelated donors. 115 patients (median age 11.4, range 0.7 – 31.6; ALL n=41, AML n=22, CML n=21, MDS/JMML n=10, NHL/HD n=4, SAA/FA n=13, SCID/WAS n=4) underwent unmanipulated allogeneic BMT (n=34) or PBSCT (n=81) from unrelated donors between 1999 and 2005. There were 88 matched- and 27 mismatched donor-recipient pairs, respectively (acc. to ALL SZT - BFM 2003 protocol). Among 88 matched pairs, 51 were 10 out of 10 HLA-allele matched (High Resolution Class I and II typed), whereas in another 37 cases 9 out of 10 alleles were matched. In the mismatched group there were 22 pairs with a 8 out of 10 allele-match and 5 pairs with 7 out of 10 allele-match, respectively. No single DRB1* mismatch was accepted. A distinct correlation between the degree of HLA-match and the risk of grades III-IV aGvHD was found (p=0.05), which occurred in 11 out of 27 pts in the mismatched group (40.7%) and in 19 out of 88 pts in the matched group (21.6%). No difference between the 2 groups was observed with regard to the incidence of extensive cGvHD. No correlation was found between the number of transplanted CD34+ cells/kg or CD3+ cells/kg and the incidence of aGvHD grades III–IV or extensive cGvHD. Median numbers of transplanted CD34+ cells/kg and CD3+ cells/kg were 7.7 × 106 (10.9 × 106) and 2.6 × 108 (3.3 × 108) respectively in the group of patients with grades III–IV aGvHD (extensive cGvHD) and 9.3 × 106 (8.8 × 106) and 3.4 × 108 (3.1 × 108) respectively in the group without these complications. Pts in the mismatched group were given slightly higher numbers of CD3+ cells/kg (4.9 vs 3.0 × 108 in the matched group) and similar numbers of CD34+ cells/kg (median 8.1 vs 8.9 × 106 in the matched group). Despite a higher incidence of severe aGvHD in the mismatched group, probability of 5-year survival was in contrast slightly better in the mismatched group (pS = 0.59 vs pS = 0.48 in the matched group, p=NS). Seventeen from 27 pts in the mismatched group remain alive (62.9%). Median follow-up of all 63 surviving pts (55%) is 21 months (range 6 – 69). Neither degree of HLA match, nor the source of stem cells (PB vs BM), nor number of transplanted CD34 (less or more than 8 × 106/kg) or CD3 cells/kg had an impact on survival. In conclusion, there seems to be no correlation between the numbers of transplanted CD34+ cells/kg and/or CD3+ cells/kg and the incidence of severe GvHD in the unrelated donor setting in children. It is safe and feasible to perform allogeneic transplants from unrelated mismatched donors, provided a detailed selection of more than 1 HLA-allele-mismatched donors is performed. This conclusion remains in contrast with the ALL SZT-BFM 2003 protocol, which requires extensive T-cell depletion in case of mismatched donor-recipient pairs. High number of transplanted CD34+ cells seems to be a positive factor influencing engraftment and survival in this constellation. There seems to be not necessary to limit the number of transplanted CD34+ cells to 6 or 8 × 106/kg, which had been previously suggested by studies performed in Europe or USA in adults.

The kinetics of the reactions of CH2Br and CH2I radicals with O2 have been studied in direct measurements using a tubular flow reactor coupled to a photoionization mass spectrometer. The radicals have been homogeneously generated by pulsed laser photolysis of appropriate precursors at 193 or 248 nm. Decays of radical concentrations have been monitored in time-resolved measurements to obtain the reaction rate coefficients under pseudo-first-order conditions with the amount of O2 being in large excess over radical concentrations. No buffer gas density dependence was observed for the CH2I + O2 reaction in the range 0.2–15 × 1017 cm−3 of He at 298 K. In this same density range the CH2Br + O2 reaction was obtained to be in the third-body and fall-off area. Measured bimolecular rate coefficient of the CH2I + O2 reaction is found to depend on temperature as k(CH2I + O2) = (1.39 ± 0.01) × 10−12 (T/300 K)−1.55 ± 0.06 cm3 s−1 (220–450 K). Obtained primary products of this reaction are I atom and IO radical and the yield of I-atom is significant. The rate coefficient and temperature dependence of the CH2Br + O2 reaction in the third-body region is k(CH2Br + O2 + He) = (1.2 ± 0.2) × 10−30 (T/300 K)−4.8 ± 0.3 cm6 s−1 (241–363 K), which was obtained by fitting the complete data set simultaneously to a Troe expression with the Fcent value of 0.4. Estimated overall uncertainties in the measured reaction rate coefficients are about ±25%.

Monosomy 7 (−7) and deletion 7q [del(7q)] are rare in childhood AML and are considered to be associated with a poor outcome. We retrospectively analyzed data on 267 children with de novo AML or RAEB-T (PB or BM blasts > 20%) with −7 or del(7q) with or without additional cytogenetic aberrations (other). Patients were diagnosed between 1985 and 2003. Karyotypes showed −7 (n=90), −7 other (n=82), del(7q) (n=22), and del(7q) other (n=73). All 24 patients with RAEB-T had −7 +/− other. The median age at diagnosis was 7.6 years (range; 0–18.1) with no difference in age distribution between cytogenetic subgroups. The most common additional cytogenetic aberrations were inv(3)/t(3;3) in 19 patients (−7, n=17; del(7q), n=2) and favorable aberrations [t(8;21), inv(16), and t(15;17)] observed in 25 patients (−7, n=3; del(7q), n=22). The 5-year overall probability of survival for the whole group was 39%, SE=3%, and survival was higher in patients with del(7q) +/− other compared with −7 +/− other (52% vs. 30%). The survival of patients with del(7q) and favorable cytogenetic aberrations (n=22) was higher than that of other patients with del(7q) (75% vs. 46%, p=0.026). There was no significant difference in survival for patients with −7 and −7 other. Complete remission (CR) was obtained in 188 patients (−7 +/− other 62%; del(7q) +/− other 88%), of these, 67 received hematopoietic stem cell transplantation (HSCT) in CR1. There was no significant difference (Mantel-Byar test) in survival for patients who received HSCT in CR1 (49%, SE= 7%) and those who received chemotherapy only (51%, SE=5%, 54% for patients surviving at least the median time to HSCT). In conclusion childhood AML subgroups −7 and del(7q) differ in their associated cytogenetic aberrations and response to therapy.