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Abstract Acute myeloid leukemia (AML) is a heterogeneous and complex disease, and treatments for this disease have not been curative for the majority of patients. In younger patients, internal tandem duplication of FLT3 (FLT3-ITD) is a common mutation for which two inhibitors (midostaurin and gilteritinib) with varied potency and specificity for FLT3 are clinically approved. However, the high rate of relapse or failed initial response of AML patients suggests that the addition of a second targeted therapy may be necessary to improve efficacy. Using an unbiased large-scale CRISPR screen, we genetically identified BCL2 knockout as having synergistic effects with an approved FLT3 inhibitor. Here, we provide supportive studies that validate the therapeutic potential of the combination of FLT3 inhibitors with venetoclax in vitro and in vivo against multiple models of FLT3-ITD-driven AML. Our unbiased approach provides genetic validation for co-targeting FLT3 and BCL2 and repurposes CRISPR screening data, utilizing the genome-wide scope toward mechanistic understanding.

This paper examines teacher education research in the field of physical education and the relationships between research, specific frameworks, and practice. The strengths and limitations of selected frameworks, through several theoretical perspectives including behavioural analysis, occupational socialization, and critical rationalization, are explored. These perspectives have demonstrated how research has influenced the teaching of physical education. Professional development, as it relates to the physical education teacher (school subject specialist) is discussed in the context of teacher development. [ABSTRACT FROM AUTHOR]

Relapse rates in high-risk neuroblastoma remain exceedingly high. The malignant cells that are responsible for relapse have not been identified, and mechanisms of therapy resistance remain poorly understood. Here, we used single nucleus RNA sequencing and bulk whole genome sequencing to identify and characterize the residual malignant persister cells that survive chemotherapy from a cohort of 20 matched diagnosis and definitive surgery tumor samples from patients treated with high-risk neuroblastoma induction chemotherapy. We show that persister cells share common mechanisms of chemotherapy escape including suppression of MYCN activity and activation of NF-κB signaling, the latter is further enhanced by cell-cell communication between the malignant cells and the tumor microenvironment. Overall, our work dissects the transcriptional landscape of cellular persistence in high-risk neuroblastoma and paves the way to the development of new therapeutic strategies to prevent disease relapse.

Background: There is limited evidence regarding the prevalence and impact of iron deficiency (ID) in children with dilated cardiomyopathy (DCM)., Methods: Retrospective single-center review of all children between 2010 and 2020 with a diagnosis of DCM and complete iron studies. ID was defined as ≥2 of ferritin <20 μg/liter, iron <9 μmol/liter, transferrin >3 g/liter, or transferrin saturation (TSat) <15%. Clinical and laboratory indices and freedom from a composite adverse event (CAE) of mechanical circulatory support (MCS), heart transplant, or death were compared between children with and without ID., Results: Of 138 patients with DCM, 47 had available iron studies. Twenty-nine (62%) were iron deficient. Children with ID were more likely to be receiving inotropes (17, 59%, p = 0.005) or invasive/noninvasive ventilation (13, 45%, p = 0.016) than those who were iron replete. They had a higher incidence of anemia (22, 76%, p = 0.004) and higher NT-proBNP (1,590 pmol/liter, IQR 456-3,447, p = 0.001). Children with ID had significantly less freedom from the CAE at 1-year (54% ± 10%), 2-years (45 ± 10), and 5-years (37% ± 11%) than those without (p = 0.011). ID and anemia were the only significant predictors of the CAE on univariate Cox regression., Conclusions: ID is highly prevalent in children with DCM. Iron studies are undermeasured in clinical practice, but ID is associated with severe heart failure (HF) and an increased risk of the CAE. The need for iron replacement therapy should be considered in children who present in HF with DCM., (Crown Copyright © 2023. Published by Elsevier Inc. All rights reserved.)

Neuroblastomas have neuroendocrine features and often show similar gene expression patterns to small cell lung cancer including high expression of delta-like ligand 3 ( DLL3 ). Here we determine the efficacy of rovalpituzumab tesirine (Rova-T), an antibody drug conjugated (ADC) with a pyrrolobenzodiazepine (PBD) dimer toxin targeting DLL3, in preclinical models of human neuroblastoma. We evaluated DLL3 expression in RNA sequencing data sets and performed immunohistochemistry (IHC) on neuroblastoma patient derived xenograft (PDX), human neuroblastoma primary tumor and normal childhood tissue microarrays (TMAs). We then evaluated the activity of Rova-T against 11 neuroblastoma PDX models using varying doses and schedules and compared anti-tumor activity to expression levels. DLL3 mRNA was differentially overexpressed in neuroblastoma at comparable levels to small cell lung cancer, as well as Wilms and rhabdoid tumors. DLL3 protein was robustly expressed across the neuroblastoma PDX array, but membranous staining was variable. The human neuroblastoma array, however, showed staining in only 44% of cases, whereas no significant staining was observed in the normal childhood tissue array. Rova-T showed a clear dose response effect across the 11 models tested, with a single dose inducing a complete or partial response in 3/11 and stable disease in another 3/11 models. No overt signs of toxicity were observed, and there was no treatment-related mortality. Strong membranous staining was necessary, but not sufficient, for anti-tumor activity. Rova-T has activity in a subset of neuroblastoma preclinical models, but heterogeneous expression in these models and the near absence of expression seen in human tumors suggests that any DLL3-targeting clinical trial should be only performed with a robust companion diagnostic to evaluate DLL3 expression for patient selection., Competing Interests: Disclosure of Potential Conflicts of Interest None.

The increasing interest in the possible role of antioxidant vitamins in many disease states means that methods of assessing vitamin intakes which are suitable for large-scale investigations are now required. The suitability of the food-frequency questionnaire, which was developed by the Medical Research Council - Cardiff Group, for determining dietary intake of antioxidant vitamins in epidemiological studies was investigated in 196 Scottish men. The validity of the dietary data was assessed by comparison with serum vitamin concentrations, and separate analyses were performed for current smokers and non-smokers. The results showed that total energy intake and the percentage of energy derived from sugar were higher in smokers, and that both dietary and serum values of vitamin C, beta-carotene and vitamin E were lower in smokers than non-smokers. After adjustment for serum lipids, energy intake and body mass index, correlation coefficients between dietary and serum vitamins C and E were similar for smokers (r 0.555 and 0.25 respectively) and non-smokers (r 0.58 and 0.32 respectively). Correlation between dietary and serum carotenes was reduced from 0.28 in non-smokers to 0.09 in smokers and correlations for retinol and total vitamin A were weakly significant only for non-smokers. The food-frequency questionnaire assigned greater than 70% of subjects correctly into the upper or lower plus adjacent tertiles of serum vitamin values, with the exception of beta-carotene and total vitamin A for smokers. Thus, the food-frequency questionnaire appeared to be an adequate tool for assigning individuals into tertiles of serum antioxidant vitamins with the main exception of beta-carotene for smokers. Marked differences do occur between the vitamins and between the smoking groups which may reflect reduced accuracy of reporting on the food-frequency questionnaire or differential absorption and metabolism of the vitamins.

Millimolar concentrations of manganese are required for maximal activation of purified galactosyltransferase (lactose synthase, EC 2.4.1.22), the enzyme that catalyzes addition of galactosyl groups to proteins and, in lactose synthesis, to glucose. To examine manganese activation of this enzyme under in vivo conditions, we studied intact, partially purified Golgi membranes from mouse mammary glands. In intact vesicles treated with the divalent cation ionophore, A23187, activation followed Michaelis-Menton kinetics with a Km of 3 microM; maximal activation was achieved below 10 microM manganese. In both detergent-solubilized and leaky vesicles the kinetics of manganese activation were consistent with the presence of two manganese-binding sites with dissociation constants about 40 microM and 20 mM. The difference is consistent with the presence in intact vesicles of an endogenous activator too large to traverse the membrane via A23187; this activator could bind to the low affinity manganese site allowing manganese or another divalent cation such as zinc to activate the enzyme at micromolar concentrations. The Km for UDP-galactose was found to be similar in the vesicular and solubilized preparations at micromolar and millimolar manganese concentrations, respectively, providing additional evidence for this hypothesis.

The volumes and spin-lattice (T1) relaxation times of breast tissues and parenchymal water content were measured non-invasively by magnetic resonance imaging (MRI) in eight healthy women during four to eight consecutive menstrual cycles. Total breast volume, and parenchymal volume, T1 relaxation time and water content were lowest between days 6 and 15. Between days 16 and 28, parenchymal volume, T1 relaxation time and water content rose sharply by 38.9%, 15.1% and 24.5%, respectively, and peaked after day 25. Within 5 days of the onset of menses, parenchymal volume fell sharply by 30.3%, while water content declined by 17.5%. Rising parenchymal volume in the second half of the menstrual cycle is not solely due to increased tissue water content and provides in vivo evidence for both growth and increased tissue fluid at this time.

Prostaglandins E1, E2 and F2 alpha were measured in duodenal secretions from two healthy, fasting male subjects. Secretions were collected by duodenal intubation prior to, and following, stimulation by secretin and cholecystokinin. Prostaglandins were measured with a radioimmunoassay technique. The ranges of concentrations found were PGE1: 2.4-110 ng/ml, PGE2: 0.004-1.51 ng/ml, and PGG2 alpha: 0.44-14.6 ng/ml.

The chromium concentration was determined in 261 samples of breast milk collected by manual expression from forty-five American women. Cr was measured in whole, liquid milk by graphite-furnace atomic absorption spectrophotometry, using the method of standard additions. The mean Cr content of the breast milk samples was 0.30 ng/ml. The range of individual values was 0.06-1.56 ng/ml and did not change significantly with duration of lactation. The Cr content of breast milk from these American women is similar to levels reported recently in milk from Finnish women.

Selenium concentrations were measured in tissue samples obtained at autopsy from 45 New Zealand residents aged 4 months to 74 yr. Materials included liver, kidney cortex, skeletal muscle, heart, lung, and spleen. Samples of liver and brain were obtained from 16 fetuses (gestational ages, 27-42 wk). Except for kidney cortex, which contained high levels, concentrations of selenium were similar to those reported in other low-selenium areas, and lower than values obtained in selenium-adequate localities.

Within-feed and between-breast differences in the concentrations of sodium, potassium, chloride, calcium, magnesium, lipid, protein, lactose, glucose, urea nitrogen, creatinine, zinc, and copper were examined in milk samples from 10 women. The average lipid content doubled in the interval from the beginning of the feed to the end. The composition of the aqueous phase of milk, as determined by the major osmotically active constituents, did not vary significantly within the feed. For these components as well as for lipid a small mid-feed sample of milk gave the same mean composition as the pooled, pumped contents of one breast suggesting that such a sample is adequate for determination of milk composition in population studies. Sporadic, inconsistent differences in the composition of the milk from the right and left breasts were observed. It is suggested that mastitis may contribute to these differences. It is recommended that samples routinely be taken from both breasts and analyzed for sodium and chloride to rule out episodes of mastitis or other local phenomena which sporadically alter milk composition.

Uptake of zinc with human milk, cow's milk, and four infant formulas investigated using a standard zinc loading test. Female subjects consumed 25 mg of zinc with the milk or formula, the amount of which was calculated to provide 5 gm of protein, after an eight-hour fast. Blood samples were taken prior to (base line) and at 30-minute intervals for three hours after consumption of zinc. Plasma zinc response was calculated as the area under the curve change in plasma zinc from the base line value, plotted against time over three hours. The plasma response with human milk was significantly greater (P less than .005) than with cow's milk and all the formulas. The response with cow's milk and a cow's milk-based formula was one third that with human milk; responses with a soy-based and two casein hydrolysate-based formulas were lower. These results are compatible with the concept, strongly suggested by indirect evidence, that the bioavailability of zinc with human milk is better than with cow's milk or infant formulas.

1. Concentrations of copper, manganese, zinc, nickel, cadmium and lead were measured in samples of liver, kidney, brain, heart, lung, skeletal muscle and vertebral bone from forty foetuses of 23-43 weeks gestation. 2. Cu concentrations in the liver were up to 100 times those in other tissues, but only those in the brain showed a significant increase with gestational age. 3. Mn concentrations were similar in all tissues; the over-all range was 0.35-9.27 microgram/g dry matter (DM). 4. Concentrations of Zn in the liver were much higher than in other tissues and decreased with gestational age, whereas levels in skeletal muscle increased. 5. In all tissues Ni concentrations were within the range 0.04-2.8 microgram/g DM and levels in kidney and muscle decreased significantly with age. 6. Cd was detected in most of the tissue samples and concentrations were within the range 0.01-0.58 microgram/g DM. 7. Concentrations of Pb, where it was detected, varied from 0.1 to 2.4 microgram/g DM in the soft tissues and from 0.4 to 4.3 microgram/g DM in the bone samples.

The concentrations of iron, copper, zinc, manganese, nickel, cadmium and lead are reported for milk foods and supplements offered to infants before the introduction of mixed feeding. Levels of most of the trace elements in samples of breast milk and homogenised cows' milk were within the ranges reported overseas. Most of the milk products should supply adequate amounts of iron, copper, manganese and nickel to the young infant although zinc intakes may be marginal. Concentrations of cadmium and lead were well below the maximum values permitted by law.

The information currently available on the levels of trace elements in New Zealand autopsy material is reviewed. Concentrations of copper, manganese, zinc, selenium and cadmium have been reported in a variety of tissues from fetuses, infants and adults, nickel and lead in some fetal tissues and adult liver, and arsenic, bismuth, cobalt, mercury, silver and thallium in adult livers. Except for Se levels, which are markedly lower, and Cu levels which are at the lower end of the range for adults, levels of trace elements are generally similar to those reported from other countries.

The intakes of milk and specific nutrients during the first 120 hours after birth were measured in 11 full-term, breast-fed infants. Infants were test weighed at all feeds using an electronic balance, and milk samples were obtained from both breasts one to three times daily. Milk was analyzed for levels of fat, protein, lactose, calcium, sodium, and potassium; energy content was calculated using the Atwater factors. The average (+/- SD) intake of milk in the first 24 hours after birth was 13 +/- 16 g/kg (range, 3 to 32 g/kg), increasing to 98 +/- 47 g/kg (50 to 163 g/kg) and 155 +/- 29 g/kg (110 to 196 g/kg) on days 3 and 5, respectively. Mean daily intakes of energy, lactose, calcium, and potassium were less than 12% of the mean day 5 intake on day 1 and less than 25% of the day 5 intake on day 2. In the first few days after birth, the nutrient intake of the solely breast-fed infant is highly variable and is frequently low.

Molybdenum and nickel were measured in 62 and 46 samples, respectively, of human milk collected from 13 women between delivery and 38 d postpartum. Trace elements were analyzed by graphite-furnace atomic absorption spectrophotometry, with standard additions to whole milk without pretreatment. Molybdenum levels (mean +/- SD) fell from 15.0 +/- 6.1 ng/mL on day 1 to an apparently constant level of 1-2 ng/mL by 1 mo. Nickel concentrations did not change with time; the overall mean was 1.2 +/- 0.4 ng/mL. Intakes of the elements by infants were calculated with test-weighing data on the fully breast-fed infants of these same mothers. By 1 mo, the average daily intake of molybdenum was 1.5 micrograms; that of nickel was 0.8 microgram.

Concentrations of Cu, Mn, Zn, and Cd were measured in 13 different tissues collected at autopsy from 55 New Zealanders, aged 1 week to 74 years. All analyses were done by atomic absorption spectrophotometry. In general, concentrations of Mn and Zn were similar to those reported elsewhere but Cu levels were slightly lower. Concentrations of Cd were low in all tissues except kidney. Median values were in accordance with those reported for other "unexposed" populations. A significant trend of increasing concentrations with age was found for Cu in cartilage, Zn in kidney cortex and medulla, and Cd in all tissues except bone, fat, and hair. Declines with age were observed for Cu in liver, aorta, and skeletal muscle, for Mn in heart, aorta, and cartilage and for Zn in lung and muscle. There were no obvious relationships between tissue trace element levels and cause of death assigned according to three groups: sudden accidental, cardiovascular, or respiratory.

Two 9-week-old, breast-fed premature infants developed acrodermatitis and hypozincemia because of low zinc content in their mothers' breast milk. All symptoms of zinc deficiency disappeared within seven days after the infants were treated orally with zinc and did not recur when zinc was discontinued after 11 months (infant 1) and three weeks (infant 3). After a subsequent term pregnancy, one of the mothers had lower breast milk zinc content (P less than .025) and greater exponential decline (P less than .025) of zinc content through 40 weeks of lactation compared with 34 control subjects. Her second infant (infant 2) had hypozincemia at 7 months of age but did not develop clinical zinc deficiency. When the mothers of the affected infants took oral zinc supplements, there was no increase in their breast milk zinc content. Zinc secretion into breast milk appears to be a controlled process that is independent of maternal zinc intake or serum zinc level. Breast milk may be low in zinc because of defective mammary secretion and this may lead to severe symptomatic zinc deficiency in premature infants.

Zinc, copper, and manganese were measured in milk samples collected longitudinally from 13 highly selected, healthy lactating women. Samples were obtained at least weekly up to 1 mo and then monthly up to 18 mo postpartum or to weaning. Zn concentrations in milk declined throughout lactation from 71.9 +/- 18.3 mumol/L (means +/- SD) at 7 d to 44.3 +/- 10.7 mumol/L at 1 mo and 7.64 +/- 4.59 mumol/L at 12 mo. Cu levels also declined from 9.76 +/- 2.05 mumol/L at 7 d to 3.46 +/- 0.79 mumol/L at 5 mo with little change thereafter. Zn and Cu were not affected by weaning. Mn concentrations declined from 67.4 +/- 23.7 nmol/L at 1 mo to approximately 36 nmol/L by 3 mo and then remained constant to 7 mo when mean levels started to rise. Gradual weaning was associated with large and irregular changes in Mn levels. Total daily outputs in milk averaged 26.0 mumol Zn, 3.9 mumol Cu, and 35 nmol Mn over the first 3 mo postpartum and 13.8 mumol Zn, 2.7 mumol Cu, and 29 nmol Mn over the second 3 mo.