Establishing a model for childhood obesity in adolescent pigs.

OBJECTIVE: Rising worldwide prevalence of obesity and metabolic diseases in children has accentuated the importance of developing prevention and management strategies. The objective of this study was to establish a model for childhood obesity using high-fat feeding of adolescent pigs, as pigs have a longer developmental period and are physiologically more similar to humans than rodents. METHODS: Crossbred pigs were fed a high-fat diet (HFD) or low-fat diet (n = 6/treatment) from postnatal day 49 to 84. On postnatal day 84, an oral glucose tolerance test was performed, jugular blood sampled to determine lipopolysaccharide levels and plasma lipids, intestinal digesta collected to characterize microbial and metabolite composition and back fat and intestinal tissue assayed for gene expression. RESULTS: Five-week HFD increased weight gain and back fat thickness, caused dyslipidaemia and impaired glucose tolerance and increased expression of genes in back fat suggesting inflammation. HFD pigs had distinct proximal colon microbiota with 48% reduction (P < 0.05) in Bacteroidetes and increased expression of pro-inflammatory genes interleukin-18 and tumour necrosis factor in ileum (P < 0.05). CONCLUSIONS: These findings indicate that adolescent pigs should be considered a suitable model for childhood obesity, because short-term HFD feeding is sufficient to induce obesity and glucose intolerance, recapitulating disease characteristics in adolescent pigs.

Acute megakaryoblastic leukemia in infants with t(1;22)(p13;q13) abnormality.

Six infants with acute megakaryoblastic leukemia and a translocation (1;22)(p13;q13) were studied. There were five female infants and one male infant, and the age at initial examination varied from 0.8 to 6.5 months (median, 2.3 months). All the patients had hepatosplenomegaly and anemia (6 to 8.3 g/dL), and four patients had thrombocytopenia (9,000 to 63,000/mm3). The bone marrow showed prominent fibrosis in five cases and reticulin fibrosis in one patient at presentation. Crush artifact often made the histologic sections difficult to interpret, but typical megakaryoblasts could be identified in the smears. Biopsy specimens of the liver and lymph node were suggestive of a nonhematopoietic malignant condition because of the cohesiveness of the tumor cells, stromal fibrosis, and the prominent sinusoidal and vascular pattern of infiltration. Immunophenotyping of peripheral blood mononuclear cells was helpful in identifying the blasts as belonging to the megakaryoblastic lineage. Using a panel of mononclonal antibodies, it was also possible to confirm the nature of the infiltration in paraffin sections and to differentiate it from other childhood small round cell tumors, especially neuroblastoma in paraffin sections (typical staining pattern: CD45-, CD43+, vW Factor, Ulex europeus I+, CD20-, CD45RO-, synaptophysin-, chromogranin-, cytokeratin-, desmin-). This special type of infantile acute leukemia can be recognized with confidence if one is aware of its clinical features, peculiar pathologic characteristics, the morphologic features and immunophenotype of the megakaryoblasts, and the unique cytogenetic abnormality.

Prevalence of myeloperoxidase gene expression in infant acute lymphocytic leukemia.

The enzyme myeloperoxidase (MPO; donor: H2O2 oxidoreductase, EC1.11.1.7) is a well-established marker of myeloid differentiation. Most myeloid leukemias express MPO enzyme activity at the light microscopic level, whereas lymphoid leukemias characteristically lack such expression. However, the diagnostic significance of MPO RNA expression or of immunohistochemically detectable MPO protein expression in leukemic blasts is unclear. We studied the prevalence and diagnostic significance of MPO RNA and protein expression in 57 cases of MPO enzyme-negative infant B-precursor acute lymphocytic leukemia (ALL), since the blast cells in this condition have been reported to show a high incidence of coexpression of myeloid-associated antigens. MPO expression was compared with other clinical and laboratory parameters. Of the cases examined, 56% showed detectable MPO expression at the RNA or protein level or both. Most positive cases showed MPO protein in many leukemic blasts, whereas a few cases showed substantial MPO protein expression in only a few blast cells. MPO expression showed no significant correlation with other markers of myeloid differentiation. Leukemic lymphoblasts in infant ALL frequently express MPO at the RNA or protein level; this expression does not imply an overall myeloid phenotype. The leukemic blasts in infant ALL may derive from an immature hematopoietic precursor cell not fully committed to lymphoid differentiation.

Myeloperoxidase gene expression in infant leukemia: a Pediatric Oncology Group Study.

A high incidence of co-expression of myeloid-associated antigens in infant B-precursor Acute Lymphocytic Leukemia (B-ALL) has been reported, but the significance of this finding is uncertain. To further assess myeloid differentiation and its prognostic significance in this disease, we investigated the frequency of myeloperoxidase (MPO) gene expression in the blast cells from 43 infants with B-ALL registered in a Pediatric Oncology Group (POG) Pilot Study for Treatment of Infant ALL, utilizing a molecular probe for detection of MPO messenger RNA (mRNA) by Northern blot hybridization and a monoclonal antibody to detect MPO-protein by immunohistochemical staining. Sufficient RNA for Northern blot was extracted from 32 bone marrow or blood samples. In two cases, MPO mRNA was determined by a reverse transcriptase-polymerase chain reaction assay and was negative in both cases. MPO-specific transcripts (MPO+) were present in 19 of 34 (56%) samples analyzed. Immunoreactive MPO protein was positive in 13 of the 20 (65%) patients studied. No correlation was found between MPO gene expression and clinical or laboratory features, karyotypic patterns or clinical outcome. The high frequency of MPO gene expression demonstrated in this study suggests that leukemogenic events in many cases of infant B-ALL appear to involve a pluripotent stem cell not yet fully committed to lymphoid differentiation.

Sequential quantitative EEG analysis in acute lymphocytic leukemia of children.

Potential brain toxicity is a major concern in the treatment of acute lymphocytic leukemia with cranial irradiation or intrathecal methotrexate. We used quantitative EEG analysis based on the Fourier transform to study 13 children at the time of diagnosis, after induction, and following consolidation which included extended intrathecal chemotherapy. None had detectable CNS infiltration by leukemia. Nonetheless, initial EEG frequencies were markedly depressed compared to expected values for age (p less than .001), and improved dramatically after induction (p less than .001). Following consolidation, EEG frequencies remained significantly lower than predicted from a control population (p less than .05). Quantitative EEG is a sensitive procedure that appears useful in assessing subtle neurologic effects of acute leukemia and its treatment.

Splenic infarction at low altitude in a child with hemoglobin S-C disease.

We describe a 15-year-old black boy with hemoglobin S-C disease living in Atlanta (altitude 1,034 ft), with no prior history of aircraft or mountain travel, who developed splenic infarction. The clinical picture was characterized by severe left upper quadrant abdominal pain, fever, splenomegaly, and hematologic and scintigraphic evidence of functional asplenia. The diagnosis was suggested by liver/spleen scintigraphy and further confirmed by ultrasonography and computerized tomography (CT) of the spleen. Treatment consisted of analgesics, intravenous fluids, and short-term antibiotic therapy. The child recovered without sequelae.

Cisplatin in children: hearing loss correlates with iris and skin pigmentation.

Pigmentation is reported to affect cisplatin-induced ototoxicity in adult humans. The hearing loss is worse in people with brown irises, than in those with blue irises. We assessed the hypothesis that cisplatin-treated children with dark irises suffer more deterioration in auditory thresholds than do those with less pigmentation. For the 19 children whose data met the requirements of this observational retrospective study, we found a weak correlation (Spearman's r = 0.50; p < 0.05) of high frequency hearing loss (at 4000 Hz) and pigmentation. Blue or hazel-eyed children averaged 2.9 dB worsening at 4000 Hz, in contrast to 14.2 dB worsening for brown or black-eyed children. Pigmentation may account for some of the individual susceptibility to cisplatin ototoxicity. We suggest that iris colour be included in future reports of cisplatin-related hearing loss.

Patient and provider perspectives on quality and health system effectiveness in a transition economy: evidence from Ukraine.

Facing a severe population health crisis due to noncommunicable diseases, Ukraine and other former Soviet republics and Eastern European countries have a pressing need for more effective health systems. Policies to enhance health system effectiveness should consider the perspectives of different stakeholder groups, including providers as well as patients. In addition, policies that directly target the quality of clinical care should be based on objective performance measures. In 2009 and 2010 we conducted a coordinated series of household and facility-level surveys to capture the perspectives of Ukrainian household members, outpatient clinic patients, and physicians regarding the country's health system overall, as well as the quality, access, and affordability of health care. We objectively measured the quality of care for heart failure and chronic obstructive pulmonary disease using CPV(®) vignettes. There was broad agreement among household respondents (79%) and physicians (95%) that Ukraine's health system should be reformed. CPV(®) results indicate that the quality of care for common noncommunicable diseases is poor in all regions of the country and in hospitals as well as polyclinics. However, perspectives about the quality of care differ, with household respondents seeing quality as a serious concern, clinic patients having more positive perceptions, and physicians not viewing quality as a reform priority. All stakeholder groups viewed affordability as a problem. These findings have several implications for policies to enhance health system effectiveness. The shared desire for health system reform among all stakeholder groups provides a basis for action in Ukraine. Improving quality, strengthening primary care, and enhancing affordability should be major goals of new health policies. Policies to improve quality directly, such as pay-for-performance, would be mutually reinforcing with purchasing reforms such as transparent payment mechanisms. Such policies would align the incentives of physicians with the desires of the population they serve.

Emperipolesis of neutrophils by dysmorphic megakaryocytes.

Neutrophil engulfment by megakaryocytes was observed within 20 to 30% of megakaryocytes from two children: one with metastatic rhabdomyosarcoma, the other with fever of unknown origin. Other cell types and neutrophil precursors were not observed within megakaryocytes. Only late megakaryocytes were involved in the process, and often these cells appeared vacuolated or degenerating at the light and electron microscope level. Ultrastructurally the engulfed neutrophils were intact and were within the open canalicular system of the megakaryocyte cytoplasm. No evidence of neutrophil granule exocytosis could be demonstrated in ultrastructural morphologic and peroxidase preparations; however, many neutrophils appeared to be endocytosing portions of the megakaryocyte cytoplasm. The phenomenon could not be transferred to normal marrow incubated with patient serum or plasma. Thus, our patients differ from previous observations of emperipolesis in: 1) the extreme frequency of the observation; 2) the selective involvement of neutrophils; and 3) the association of the anomaly with dysmorphic and/or disrupted megakaryocytes. These observations are consistent with a neutrophil response to altered and/or injured megakaryocytes.

Plasma dopa and catecholamines in the diagnosis and follow-up of children with neuroblastoma.

Plasma dopa and the catecholamines--dopamine, norepinephrine, and epinephrine--were assayed by a radioenzymatic method in 15 children with active neuroblastoma and in eight others without evidence of disease to assess the value of these determinations in the diagnosis and management of the tumor. Thirty-four children with solid tumors and hemopoietic malignancies served as our controls. Elevated plasma dopa levels were observed in 13 children with active neuroblastoma (87%); dopamine and norepinephrine were elevated in 1/4 of these patients. In the group of children with neuroblastoma without evidence of disease, dopa and catecholamine levels were within the range observed in the controls. Total urinary catecholamines, homovanillic acid (HVA) and/or vanilmandelic acid (MVA) were elevated in 11 of the 15 (73%) neuroblastoma patients with active disease. While serial plasma dopa determinations correlated with the course of the disease in practically all patients and thus were useful in their follow-up, the catecholamines were of limited value in assessing tumor status. Our results suggest that plasma dopa, assayed by a radioenzymatic method, may be more reliable than the traditional urinary catecholamine determinations in the diagnosis of neuroblastoma, and it appears useful in the management of this disease.

Use of VP-16-213 in the treatment of familial erythrophagocytic lymphohistiocytosis.

Five patients with familial erythrophagocytic lymphohistiocytosis (FEL), aged 6 weeks to 3 years, were treated with VP-16-213. The drug dosage ranged from 100 to 200 mg/m2 administered biweekly until remission was achieved, and then at 1- to 3-week intervals as maintenance therapy. Intrathecal methotrexate was given to two patients with central nervous system involvement. All patients attained remission. Systemic relapses often ensued in all patients when VP-16-213 was delayed because of myelosuppression, or after attempts to lengthen the treatment interval, but they initially responded again to a more intensive chemotherapy schedule. To date, four of the patients died from disseminated disease and terminal infections 15 to 20 months from the time of diagnosis. One child is alive and well 20 months from diagnosis. Three of the dead children had become refractory to the drug. Our observations show that VP-16-213 induces remissions and prolongs survival in FEL. However, since the patients eventually become refractory to the drug and die of the disease, additional forms of therapy are required to improve the outlook of affected children.

Adjuvant chemotherapy for advanced nasopharyngeal carcinoma in childhood.

Seven children with advanced nasopharyngeal carcinoma younger than 20 years of age diagnosed between 1975 and 1986 (inclusive) were treated with a uniform adjuvant chemotherapy regimen, which consisted of vincristine (1.5 mg/m2; day 1), doxorubicin (45 mg/m2; day 1), 5-fluorouracil (8 mg/kg; days 1 through 5), and cyclophosphamide (7 mg/kg; days 1 through 5). This combination chemotherapy was given for 12 to 24 months after completion of radiation therapy. The radiation doses to the primary sites ranged from 6000 cGy to a maximum of 6800 cGy. The radiation doses for neck prophylaxis ranged from 4500 cGy to a total of 5000 cGy. Involved sites were irradiated to at least an additional boost of 1000 cGy. One patient had an external dose 6000 cGy to the primary site boosted with brachytherapy of 3000 cGy at the surface of an ovoid. After chemotherapy myelosuppression occurred in all patients and was tolerable. All seven patients are surviving, six disease-free, for 22 months to 12 years (median, 4 years). This study suggests that the combination of radiation therapy and chemotherapy as used here has acceptable toxicity and is effective and further suggests that children with nasopharyngeal carcinoma, even in its advanced stage at diagnosis, may be curable.

Chemotherapy for patients with recurrent or refractory neuroblastoma: a POG Phase II study.

PURPOSE: The purpose of this study was to evaluate the efficacy and toxicity of three different salvage regimens (Rx) in children with recurrent or refractory neuroblastoma. PATIENTS AND METHODS: Forty-six children with recurrent or refractory neuroblastoma received treatment according to one of three regimens: Rx 1 (five patients), high-dose cisplatin (HDP) (200 mg/m2) with concurrent sodium thiosulfate (STS) (9.9 g/m2) as a nephroprotectant and etoposide (VP-16) (200 mg/m2/day for 3 days); Rx 2 (22 patients), high-dose carboplatin (HD-CBDCA) (500 mg/m2/day for 2 days) and VP-16 (100 mg/m2/day for 3 days); Rx 3 (19 patients), ifosfamide (1.5 g/m2/day for 3 days) followed by CBDCA (400 mg/m2) on day 4. Chemotherapy was administered every 3-4 weeks. Responses were assessed following four courses with or without surgery. Patients achieving less than a partial response (PR) on their primary treatment were crossed over to the next regimen (i.e., Rx 1 --> Rx 2 <--> Rx 3). RESULTS: Rx 1 was ended early owing to grade 4 nephrotoxicity in two patients following their first course. Ten of 22 evaluated patients (45%) primarily (n = 19) or secondarily (n = 3) treated by Rx 2 responded [five complete response (CR) and five PRs]. Nine of the 23 evaluated patients (39%) on Rx 3 as primary (n = 18) or secondary (n = 5) treatment responded (one CR and eight PRs). Grades 3-4 neutropenia and thrombocytopenia occurred after 80% and 50% of courses administered on Rx 2 and Rx 3, respectively. Central venous line infections were the most commonly documented infections on these regimens. CONCLUSIONS: Rx 2 and Rx 3 are active combinations in patients with recurrent or refractory neuroblastoma and are associated with manageable toxicity. HDP administered as a short i.v. infusion with concurrent STS infusion cannot be safely given to children with neuroblastoma pretreated with cisplatin.

Effective adjuvant chemotherapy for advanced nasopharyngeal carcinoma in children: a final update of a long-term prospective study in a single institution.

PURPOSE: The purpose of this study was to determine the efficacy and toxicity of a doxorubicin/cyclophosphamide-based chemotherapy and local radiation therapy in children with locally advanced or metastatic nasopharyngeal carcinoma (NPC). PATIENTS AND METHODS: Twelve patients aged 6 to 20 years old were treated with a chemotherapy regimen comprised of vincristine (1.5 mg/m2) and doxorubicin (45 mg/m2) on day 1 and cyclophosphamide (210 mg/m2) and 5-fluorouracil (240 mg/m2) on days 1 to 5. Chemotherapy was administered every 3 weeks for 1 to 2 years. Radiotherapy to the primary site (59 to 68 Gy) and to the neck (59 to 66 Gy) was given before or after 2 to 4 courses of chemotherapy. RESULTS: All patients achieved a complete response 4 to 16 months from the start of therapy (median 7 months). Nine patients have remained tumor free from 2 to 21 years (median 11 years) from diagnosis. One child was lost to follow-up and one died of tuberculosis; both were disease-free. One child developed a secondary osteosarcoma in the left mandible. Chemotherapy caused grade 4 neutropenia and thrombocytopenia in four patients. There were no therapy-related deaths and the most common late effect of therapy was neck fibrosis, which was observed in all patients. We conclude that the chemotherapy and radiotherapy regimen used in this study is highly effective for children and adolescents with locally advanced NPC and is associated with tolerable toxicity.

Acute myeloid leukemia (AML) in Down's syndrome is highly responsive to chemotherapy: experience on Pediatric Oncology Group AML Study 8498.

The treatment of acute myeloid leukemia (AML) in children with Down's syndrome (DS) has engendered considerable controversy. Because of the concerns for toxicity and increased rate of infections, treatment approaches varied considerably in the past with mixed results. However, experience on the recently completed Pediatric Oncology Group (POG) 8498 AML study suggests that DS children with AML constitute a distinct subgroup that responds well to therapy. Twelve of 285 children on POG 8498 (protocol for newly diagnosed AML) had DS. Children with DS and AML were predominantly male (9 of 12) and were quite younger at diagnosis (< 24 months in 10). The white blood cell count was less than 50 x 10(3)/microL in all 12 and French-American-British types M6 and M7 were frequent (5 of 12). An abnormal cytogenetic marker, in addition to constitutional trisomy 21, was present in 9 of 12 and involved chromosome 8 in 4 of 9. All cases studied (n = 5) were positive for myeloid cell surface markers (CD33, CD13, or CD11b) and, interestingly, were also positive for the CD7 antigen. Chemotherapy included daunorubicin, cytarabine (Ara-C), and 6-thioguanine for remission induction and featured high-dose Ara-C (3 g/m2 per dose) with or without L-asparaginase early in remission. Compared with children without DS, children with DS had a superior event-free survival (EFS at 4 years 100% v 28% +/- 6.2%; P = .003). The EFS remained superior even when compared with non-DS children less than 2 years of age with a white blood cell count less than 10 x 100,000/microL (100% v 48% +/- 17.3%; P = .01).

Natural killer cells in children with malignant solid tumors. Effect of recombinant interferon-alpha and interleukin-2 on natural killer cell function against tumor cell lines.

Natural killer (NK) cells and NK cell activity were determined in three groups (newly diagnosed [n = 21], on therapy [n = 21], and off therapy [n = 18]) of children with various types of malignant solid tumors and in a control group (n = 26) by means of Leu-7 and Leu-11b monoclonal antibodies and a 4-hour 51Cr-release assay, respectively. The erythroleukemia cell line K562 was used as a target cell. The newly diagnosed group included eight patients with localized disease (Stage I-II), ten with bulky but nonmetastatic disease (Stage III), and three with metastases (Stage IV). The mean percent of NK cell activity in the newly diagnosed group was significantly higher than that of the control group. Children with Stage III tumors at diagnosis had higher mean NK cell function than those with Stage I-II and Stage IV. On therapy patients had significantly fewer NK cells and lower NK cell cytotoxicity than those in the other groups studied. We also studied the following: (1) the in vitro effect of recombinant interferon-alpha (rIFN-alpha) and recombinant interleukin-2 (rIL-2) on NK cell function of peripheral blood lymphocytes (PBL) from children with solid malignancies; and (2) the susceptibility of neuroblastoma-derived (CHP-126 and SKNSH) and rhabdomyosarcoma-derived (A-204) cell lines to NK cell lysis. Both rIFN-alpha and rIL-2 enhanced NK cell activity of PBL from children with malignancies and healthy children against K562 and solid tumor cell lines. The enhancing effect or rIL-2 was greater than that of rIFN-alpha. CHP-126 and SKNSH cell lines were susceptible to NK cell lysis mediated by the PBL of children with neuroblastoma and the control group. The A-204 cell line was less sensitive than K562 to NK cell cytotoxicity. Our results suggest a potential therapeutic role for both cytokines in the treatment of malignant solid tumors of childhood.

Clinical features and outcome in childhood T-cell leukemia-lymphoma according to stage of thymocyte differentiation: a Pediatric Oncology Group Study.

The immunophenotypes of lymphoblasts from children with newly diagnosed T-cell acute lymphoid leukemia (T-ALL, n = 101) or T-cell non-Hodgkin lymphoma (T-NHL, n = 31) were analyzed to correlate stage of thymocyte differentiation with clinical features and outcome. The 67 boys and 34 girls with T-ALL were 1 month to 18 years old (median, 8 years) with leukocyte counts ranging from 2 to 810 x 10(9)/L (median, 55 x 10(9)/L). Eighteen of these patients were black, and 70 had a mediastinal mass. Twenty-six boys and five girls with a median age of 9 years (range, 1 to 20 years) had T-NHL. Seven of these patients were black, and 24 had a mediastinal mass. The distributions of thymocyte developmental stages (early [CD7+], intermediate [CD1+ and/or CD4+ and/or CD8+], and mature [CD3+]) in cases of T-ALL and T-NHL were significantly different: 34%, 43%, and 23% v 6%, 62%, and 32% (P = .02). A comparison of the patients' clinical features according to the maturational stage of thymocytes failed to disclose significant differences in the majority of characteristics studied. However, patients with mature-stage T-NHL, with or without the addition of subjects with mature-stage T-ALL, were less likely to have a mediastinal mass (P = .02 for both comparisons). Those with intermediate-stage T-cell malignancy (T-ALL and T-NHL combined) were the subgroup most likely to have a mediastinal mass (P = .01). Response to remission induction therapy was significantly worse in the T-ALL subgroup with an early-stage phenotype: a failure rate of 21% v 0% and 6% for the two more differentiated phenotypic subgroups (P = .007). Event-free survival was not affected by thymocyte maturational stage in cases of either T-ALL or T-NHL. Despite evidence of clinical heterogeneity among the maturational stages of T-cell malignancies in children, these developmental subdivisions do not appear to be critical determinants of outcome once remission is achieved. We conclude that such phenotypes need not be included in the stratification plans for clinical trials using common induction treatment.