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Abstract Background In London and the rest of the UK, diseases associated with poor diet, inadequate physical activity and mental illness account for a large proportion of area based health inequality. There is a lack of evidence on interventions promoting healthier behaviours especially in marginalised populations, at a structural or ecological level and utilising a community development approach. The Well London project financed by the Big Lottery 'Wellbeing' Fund and implemented by a consortium of London based agencies led by the Greater London Authority and the London Health Commission is implementing a set of complex interventions across 20 deprived areas of London. The interventions focus on healthy eating, healthy physical activity and mental health and wellbeing and are designed and executed with community participation complementing existing facilities and services. Methods/Design The programme will be evaluated through a cluster randomised controlled trial. Forty areas across London were chosen based on deprivation scores. Areas were characterised by high proportion of Black and Minority Ethnic residents, worklessness, ill-health and poor physical environments. Twenty areas were randomly assigned to the intervention arm of Well London project and twenty 'matched' areas assigned as controls. Measures of physical activity, diet and mental health are collected at start and end of the project and compared to assess impact. The quantitative element will be complemented by a longitudinal qualitative study elucidating pathways of influence between intervention activities and health outcomes. A related element of the study investigates the health-related aspects of the structural and ecological characteristics of the project areas. The project 'process' will also be evaluated. Discussion The size of the project and the fact that the interventions are 'complex' in the sense that firstly, there are a number of interacting components with a wide range of groups and organisational levels targeted by the intervention, and secondly, a degree of flexibility or tailoring of the intervention, makes this trial potentially very useful in providing evidence of the types of activities that can be used to address chronic health problems in communities suffering from multiple deprivation. Trial Registration Current Controlled Trials ISRCTN68175121

Abstract Background School-based breakfast provision is increasingly being seen as a means of improving educational performance and dietary behaviour amongst children. Furthermore, recognition is growing that breakfast provision offers potential as a means of addressing social inequalities in these outcomes. At present however, the evidence base on the effectiveness of breakfast provision in bringing about these improvements is limited. Methods/Design This paper describes the research design of a large scale evaluation of the effectiveness of the Welsh Assembly Government's Primary School Free Breakfast Initiative. A cluster randomised trial, with school as the unit of randomisation was used for the outcome evaluation, with a nested qualitative process evaluation. Quantitative outcome measures included dietary habits, attitudes, cognitive function, classroom behaviour, and school attendance. The study recruited 111 primary schools in Wales, of which 56 were randomly assigned to control condition and 55 to intervention. Participants were Year 5 and 6 students (aged 9–11 years) in these schools. Data were collected for all 111 schools at each of three time points: baseline, 4 month and 12 month follow-up. This was achieved through a repeated cross-sectional survey of approximately 4350 students on each of these occasions. Of those students in Year 5 at baseline, 1975 provided data at one or both of the follow-ups, forming a nested cohort. The evaluation also included a nested process evaluation, using questionnaires, semi-structured interviews and case studies with students, school staff, and local authority scheme coordinators as key informants. Discussion An overview of the methods used for the evaluation is presented, providing an example of the feasibility of conducting robust evaluations of policy initiatives using a randomised trial design with nested process evaluation. Details are provided of response rates and the flow of participants. Reflection is offered on methodological issues encountered at various stages through the course of the study, focusing upon issues associated with conducting a randomised trial of a government policy initiative, and with conducting research in school settings. Trial registration Current Controlled Trials ISRCTN18336527

We report here the case of an infant who developed life-threatening rigid-chest syndrome after receiving an accidental overdose of methadone. The child responded to narcotic reversal. Pediatric physicians should be aware of this possible complication. [ABSTRACT FROM AUTHOR]

We investigated whether a service-planning document outlining recommendations for what providers should address in treatment (i.e., targets) and the associated clinical techniques they should employ (i.e., practices) influenced the targets and practices that providers reported actually implementing during the subsequent treatment episode. Participants included 94 youths ages 4 to 17 (M = 13.57, SD = 3.59) who received community-based mental health services from the Hawai'i Child and Adolescent Mental Health Division. Data on targets and practices were compared across initial Mental Health Treatment Plans and Monthly Treatment and Progress Summaries. Data were analyzed using two-level, generalized mixed effects models with two-way cross-classification or linear mixed effects models. Providers were more likely to report the use of targets and practices in treatment if they were included within the treatment plan. In addition, the more closely targets addressed during treatment followed the recommended targets from the treatment plan, the more closely implemented practices followed the recommended practices listed in the treatment plan. Furthermore, as providers shifted their focus to different targets, a shift in their use of practices was also evident over time. Last, practices for which there is demonstrated efficacy for particular targets were more likely to be used. Service planning documents appear to help organize care; however, results also suggest possible limitations to the current system. These findings highlight potential areas for improvement in planning and care delivery.

The Revised Child Anxiety and Depression Scale-Parent Version (RCADS-P) is a parent-report questionnaire of youth anxiety and depression with scales corresponding to the DSM diagnoses of separation anxiety disorder, social phobia, generalized anxiety disorder, panic disorder, obsessive-compulsive disorder, and major depressive disorder. The RCADS-P was recently developed and has previously demonstrated strong psychometric properties in a clinic-referred sample (Ebesutani et al., Journal of Abnormal Child Psychology 38, 249-260, 2010b). The present study examined the psychometric properties of the RCADS-P in a school-based population. As completed by parents of 967 children and adolescents, the RCADS-P demonstrated high internal consistency, test-retest reliability, and good convergent/divergent validity, supporting the RCADS-P as a measure of internalizing problems specific to depression and five anxiety disorders in school samples. Normative data are also reported to allow for the derivation of T-scores to enhance clinicians' ability to make classification decisions using RCADS-P subscale scores.

Objective: To estimate nursing workload from the patient acuity level (PAL) assigned to patients in a pediatric intensive care unit (PICU) and to determine its influence on unplanned extubations., Design: Prospective cohort study., Setting: The 19-bed PICU of an urban, university-affiliated, tertiary children's hospital., Patients: All patients admitted to the PICU., Interventions: None., Measurements and Main Results: The study encompassed 2,193 nursing shifts and 1,919 admissions to the PICU over 24 months. The shift census averaged 12.0 patients (range 5-18) and was staffed by 9.4 nurses (range 4-16) for an average patient/nurse ratio of 1.3 +/- 0.2. Patients were assigned a PAL of 1-7 based on a classification system derived from time studies of 12 general nursing tasks. The total PALs per shift divided by the number of nursing staff yielded an average assignment of 5.8 +/- 0.7 PALs. Forty unplanned extubations (0.76 unplanned extubations/100 ventilator days) were observed during the study period. Logistic regression revealed positive associations between unplanned extubations and patient/nurse ratio (p = .03) and the shift PAL/nurse ratio (p = .01). The likelihood of an unplanned extubation when nurses covered >6.3 PALs was 3.8 times higher than during those shifts when they covered <5.3 PALs., Conclusions: The likelihood of an unplanned extubation increased with higher patient/nurse and patient acuity/nurse ratios. Successful interventions to reduce the frequency of this medical error may need to address both nurse demand methodology and workload.

In peanuts, a mechanism of resistance to fungal infection is reportedly due to the synthesis of stilbene phytoalexins, which are antibiotic, low molecular weight metabolites. The phytoalexin-associated response of different peanut genotypes to exogenous invasion in the field has not been investigated and may be useful for breeding resistant peanut cultivars. Five peanut genotypes, Georgia Green, Tifton 8, C-99R, GK-7 High Oleic, and MARC I, which differ in resistance to major peanut diseases, were investigated for their ability to produce phytoalexins under field conditions in South Georgia in 2001 and 2002. Five known peanut phytoalexins, trans-resveratrol, trans-arachidin-1, trans-arachidin-2, trans-arachidin-3, and trans-3'-isopentadienyl-3,5,4'-trihydroxystilbene, were quantitated. The phytoalexins were measured in peanuts of different pod maturity (yellow, orange, brown, and black) with or without insect pod damage (externally scarified or penetrated). Kernels from insect-damaged pods of C-99R and Tifton 8 genotypes had significantly higher concentrations of phytoalexins than other genotypes. The same genotypes were the most resistant to tomato spotted wilt virus and late leaf spot, while MARC I, which is highly susceptible to these diseases, produced very low concentrations of phytoalexins. However, there was no significant difference in phytoalexin production by undamaged peanut pods of all tested genotypes. trans-Arachidin-3 and trans-resveratrol were the major phytoalexins produced by insect-damaged peanuts. In damaged seeds, the concentrations of trans-3'-isopentadienyl-3,5,4'-trihydroxystilbene were significantly higher in Tifton 8 as compared to other genotypes. There was an association between total phytoalexin production and published genotype resistance to major peanut diseases. Stilbene phytoalexins may be considered potential chemical markers in breeding programs for disease-resistant peanuts.

Preharvest aflatoxin contamination has been identified by the peanut industry as a serious issue in food safety and human health because of the carcinogenic toxicity. Drought stress is the most important environmental factor exacerbating Aspergillus infection and aflatoxin contamination in peanut. The development of drought-tolerant peanut cultivars could reduce aflatoxin contamination and would represent a major advance in the peanut industry. In this study, we identified a novel PLD gene in peanut (Arachis hypogaea), encoding a putative phospholipase D (PLD, EC 3.1.4.4). The completed cDNA sequence was obtained by using the consensus-degenerated hybrid oligonucleotide primer strategy. The deduced amino acid sequence shows high identity with known PLDs, and has similar conserved domains. The PLD gene expression under drought stress has been studied using four peanut lines: Tifton 8 and A13 (both drought tolerant) and Georgia Green (moderate) and PI 196754 (drought sensitive). Northern analysis showed that PLD gene expression was induced faster by drought stress in the drought-sensitive lines than the drought tolerance lines. Southern analysis showed that cultivated peanut has multiple copies (3 to 5 copies) of the PLD gene. These results suggest that peanut PLD may be involved in drought sensitivity and tolerance responses. Peanut PLD gene expression may be useful as a tool in germplasm screening for drought tolerance.

ABSTRACT Infection of peanut (Arachis hypogaea) seed by Aspergillus flavus and A. parasiticus is a serious problem that can result in aflatoxin contamination in the seed. Breeding resistant cultivars would be an effective approach to reduce aflatoxin accumulation. The objective of this study was to investigate the expression of the pathogenesis-related (PR) protein beta-1,3-glucanase and the isoform patterns in peanut seed inoculated with A. flavus. Peanut genotypes GT-YY9 and GT-YY20 (both resistant to A. flavus infection) and Georgia Green and A100 (both susceptible to A. flavus infection) were used in this study. The activities of beta-1,3-glucanase were similar in the uninfected seed of all genotypes, but increased significantly in the resistant genotypes after inoculation in comparison with the susceptible genotypes. An in-gel (native polyacrylamide gel electrophoresis [PAGE]) enzymatic activity assay of beta-1,3-glucanase revealed that there were more protein bands corresponding to beta-1,3-glucanase isoforms in the infected seed of resistant genotypes than in the infected seed of susceptible genotypes. Both acidic and basic beta-1,3-glucanase isoforms were detected in the isoelectric focusing gels. Thin-layer chromatography analysis of the hydrolytic products from the reaction mixtures of the substrate with the total protein extract or individual band of native PAGE revealed the presence of enzymatic hydrolytic oligomer products. The individual bands corresponding to the bands of beta-1,3-glucanase isoforms Glu 1 to 5 were separated on the sodium dodecyl sulfate-PAGE, resulting in two bands of 10 and 13 kDa, respectively. The sequences of fragments of the 13-kDa major protein band showed a high degree of homology to conglutin, a storage protein in peanut seed. Conglutin is reported as a peanut allergen, Ara h2. Our data provide the first evidences for peanut having beta-1,3-glucanase activities and the association with the resistance to A. flavus colonization in peanut seed. We have not directly demonstrated that conglutin has beta-1,3-glucanase activity.

ABSTRACT Late leaf spot disease caused by Cercosporidium personatum is one of the most destructive foliar diseases of peanut (Arachis hypogaea) worldwide. The objective of this research was to identify resistance genes in response to leaf spot disease using microarray and real-time polymerase chain reaction (PCR). To identify transcripts involved in disease resistance, we studied the gene expression profiles in two peanut genotypes, resistant or susceptible to leaf spot disease, using cDNA microarray containing 384 unigenes selected from two expressed sequenced tag (EST) cDNA libraries challenged by abiotic and biotic stresses. A total of 112 spots representing 56 genes in several functional categories were detected as up-regulated genes (log(2) ratio > 1). Seventeen of the top 20 genes, each matching gene with known function in GenBank, were selected for validation of their expression levels using real-time PCR. The two peanut genotypes were also used to study the functional analysis of these genes and the possible link of these genes to the disease resistance trait. Microarray technology and real-time PCR were used for comparison of gene expression. The selected genes identified by microarray analysis were validated by real-time PCR. These genes were more greatly expressed in the resistant genotype as a result of response to the challenge of C. personatum than in the susceptible genotype. Further investigations are needed to characterize each of these genes in disease resistance. Gene probes could then be developed for application in breeding programs for marker-assisted selection.

In the United States, insecticide is used extensively in the production of sweet corn due to consumer demand for zero damage to ears and to a sweet corn genetic base with little or no resistance to ear-feeding insects. Growers in the southern United States depend on scheduled pesticide applications to control ear-feeding insects. In a study of quantitative genetic control over silk maysin, AM-maysin (apimaysin and methoxymaysin), and chlorogenic acid contents in an F2 population derived from GE37 (dent corn, P1A1) and 565 (sh2 sweet corn, p1a1), we demonstrate that the P1 allele from field corn, which was selected against in the development of sweet corn, has a strong epistatic interaction with the a1 allele in sh2 sweet corn. We detected that the p1 gene has significant effects (P < 0.0001) not only on silk maysin concentrations but also on AM-maysin, and chlorogenic acid concentrations. The a1 gene also has significant (P < 0.0005) effects on these silk antibiotic chemicals. Successful selection from the fourth and fifth selfed backcrosses for high-maysin individuals of sweet corn homozygous for the recessive a1 allele (tightly linked to sh2) and the dominant P1 allele has been demonstrated. These selected lines have much higher (2 to 3 times) concentrations of silk maysin and other chemicals (AM-maysin and chlorogenic acid) than the donor parent GE37 and could enhance sweet corn resistance to corn earworm and reduce the number of applications of insecticide required to produce sweet corn.

Performance of experimental Bacillus thuringiensis (Bt) MON events alone and pyramided with MON810 were evaluated over 3 yr in Georgia and Alabama. Ability of events to prevent whorl defoliation by the fall armyworm, Spodoptera frugiperda (J. E. Smith), and natural ear feeding damage by the corn earworm, Helicoverpa zea (Boddie) was assessed. In each year, near-isogenic hybrids with novel single transformation events and crosses pyramided with the MON810 event were compared with the standard single MON810 event and nontransformed susceptible control. Events were tested for resistance to whorl damage by manual infestations of fall armyworm and ear damage by natural infestations of corn earworm. All Bt events tested reduced fall armyworm whorl damage ratings per plant compared with the susceptible hybrid. All Bt treatments also had considerably less ear infestation and damage by corn earworm compared with the nontransgenic isoline. The MON841, MION849, and MON851 events reduced ear damage by H. zea but were not as effective as other novel events and were not advanced for further testing after the 1999 season. Pyramiding events compared with single events did not improve control of fall armyworm whorl damage, but they generally did prevent more ear damage by corn earworm. The MON84006 event singly and pyramided with MON810 had superior control of whorl-stage damage by S. frugiperda and ear damage by H. zea compared with MON810. Deployment of new events and genes could provide additional tools for managing the potential for insect resistance to Bt toxins. Furthermore, improved control of whorl and ear infestations by H. zea and S. frugiperda would increase the flexibility of planting corn, Zea mays L., and permit double cropping of corn in areas where these pests perennially reach damaging levels.

Host-plant resistance is an efficient, economical and environmentally benign approach used to manage many pests and diseases of agricultural crops. After nearly a century of research, the resources and tools have become more refined, but the basic tasks in breeding for resistance have not changed. Resistance must be identified, incorporated into elite germplasm, and deployed in a form useful to growers. In some instances, biotechnology has expedited this process through incorporating a foreign gene(s) for resistance into elite germplasm. The USDA Agricultural Research Service (ARS) has made significant contributions in the development of germplasm with resistance to insects, nematodes and plant diseases. Because resistant plant varieties are an essential component of sustainable production systems, ARS is committed to developing techniques and germplasm to help meet this goal.

Objective: To find a body measurement that would serve as an index for determining the length of femoral venous catheter to be inserted to achieve a position near the right atrium., Methods: A candidate index measurement was chosen, and radiographic measurements of routine femoral venous catheter placements were compared with the placement that may have resulted from use of the index in a group of patients. In a subsequent group, the candidate index was used to choose catheter insertion length, the accuracy of which was again evaluated from routine placement radiographs., Results: The first series of radiographic measurements predicted that use of the sternal-umbilical-puncture (SUP) index would result in acceptable and accurate catheter tip placement. This was confirmed in the second group of patients in which 11 of 12 catheter tips were within 1 cm of the target position., Conclusions: The use of the SUP index ensures acceptable accuracy in estimating the required insertion length of femoral catheter when tip placement near the right atrium is the clinical goal.

Aflatoxin, produced by Aspergillus flavus, is one of the most toxic and carcinogenic substances known and contaminates many agricultural commodities such as corn, peanuts, cottonseed, and tree nuts. The challenge to breeders/plant pathologists is to identify lines that have resistance to aflatoxin production. Maize population GT-MAS:gk has been identified and released as a germplasm with resistance to aflatoxin contamination. In the present study, we assessed genetic divergence in the GT-MAS:gk population using restriction fragment length polymorphism (RFLP) DNA markers to survey 11 selfed inbred lines and conducted field evaluations for the dissimilarities in aflatoxin production among these inbred lines in comparison with a sister population, GT-MAS:pw.nf. The 11 selfed inbred lines were assayed for DNA polymorphism using 113 RFLP markers in 10 linkage groups covering 1,518.2 centimorgans (cM; unit of gene or chromosome size). Considerable variation among the inbreds was detected with RFLP markers, of which 42 probe-enzyme combinations gave 102 polymorphic bands. Cluster analysis based on genetic similarities revealed associations and variations among the tested lines. Three polymorphic groups were distinguished by cluster analysis. Two years of field evaluation data showed that aflatoxin concentrations among the lines were significantly different in both years (P < 0.001). Maturity data were also different. Thus, this study demonstrates that the maize population GT-MAS:gk is heterogeneous and that individuals may be different in resistance to A. flavus infection and aflatoxin production. Therefore, the most resistant lines should be inbred to increase homogeneity, and resistance should be confirmed through progeny testing.

Study Objective: To determine whether adding IV theophylline to an aggressive regimen of inhaled and IV beta-agonists, inhaled ipratropium, and IV methylprednisolone would enhance the recovery of children with severe status asthmaticus admitted to the pediatric ICU (PICU)., Design: A prospective, randomized, controlled trial. Asthma scoring was performed by investigators not involved in treatment decisions and blinded to group assignment., Setting: The PICU of an urban, university-affiliated, tertiary-care children's hospital., Patients: Children with a diagnosis of status asthmaticus who were admitted to the PICU for < or = 2 h and who were in severe distress, as indicated by a modified Wood-Downes clinical asthma score (CAS) of > or = 5., Interventions: All subjects initially received continuous albuterol nebulizations; intermittent, inhaled ipratropium; and IV methylprednisolone. The theophylline group was also administered infusions of IV theophylline to achieve serum concentrations of 12 to 17 microg/mL. A CAS was tabulated twice daily., Measurements and Results: Forty-seven children (median age, 8.3 years; range, 13 months to 17 years) completed the study. Twenty-three children received theophylline. The baseline CASs of both groups were similar and included three subjects receiving mechanical ventilation in each group. All subjects receiving mechanical ventilation and theophylline were intubated before drug infusion. Among the 41 subjects who were not receiving mechanical ventilation, those receiving theophylline achieved a CAS of < or = 3 sooner than control subjects (18.6 +/- 2.7 h vs 31.1 +/- 4.5 h; p < 0.05). Theophylline had no effect on the length of PICU stay or the total incidence of side effects. Subjects receiving theophylline had more emesis (p < 0.05), and control patients had more tremor (p < 0.05)., Conclusions: Theophylline safely hastened the recovery of children in severe status asthmaticus who were also receiving albuterol, ipratropium, and methylprednisolone. The role of theophylline in the management of asthmatic children in impending respiratory failure should be reexamined.

Maysin, a C-glycosylflavone in maize silk, has insecticidal activity against corn earworm, Helicoverpa zea (Boddie), larvae. Sweet corn, Zea mays L., is a vulnerable crop to ear-feeding insects and requires pesticide protection from ear damage. This study was conducted to identify maize chromosome regions associated with silk maysin concentration and eventually to transfer and develop high silk maysin sweet corn lines with marker-assisted selection (MAS). Using an F2 population derived from SC102 (high maysin dent corn) and B31857 (low maysin sh2 sweet corn), we detected two major quantitative trait loci (QTL). It was estimated that 25.6% of the silk maysin variance was associated with segregation in the genomic region of npi286 (flanking to p1) on chromosome 1S. We also demonstrated that a1 on chromosome 3L had major contribution to silk maysin (accounted for 15.7% of the variance). Locus a1 has a recessive gene action for high maysin with the presence of functional p1 allele. Markers umc66a (near c2) and umc105a on chromosome 9S also were detected in this analysis with minor contribution. A multiple-locus model, which included npi286, a1, csu3 (Bin 1.05), umc245 (Bin 7.05), agrr21 (Bin 8.09), umc105a, and the epistatic interactions npi286 x a1, a1 x agrr21, csu3 x umc245, and umc105a x umc245, accounted for 76.3% of the total silk maysin variance. Tester crosses showed that at the a1 locus, SC102 has functional A1 alleles and B31857 has homozygous recessive a1 alleles. Individuals of (SC102 x B31857) x B31857 were examined with MAS and plants with p1 allele from SC102 and homozygous a1 alleles from B31857 had consistent high silk maysin. Marker-assisted selection seems to be a suitable method to transfer silk maysin to sweet corn lines to reduce pesticide application.

Objective: To describe the effects of inhaled nitric oxide on oxygenation and ventilation in patients with acute, hypoxic respiratory failure and to characterize those who respond to low doses with a significant improvement in PaO2., Design: Prospective dose response trial of inhaled nitric oxide. Patients who demonstrated a > or =15% improvement in PaO2 were randomized to receive conventional mechanical ventilation with or without prolonged inhaled nitric oxide., Setting: Pediatric intensive care unit of a tertiary care children's hospital serving as a regional referral center for respiratory failure., Patients: Pediatric patients with an acute parenchymal lung disease requiring mechanical ventilation, an F(IO2) of > or =0.5, a positive end-expiratory pressure of > or =7 cm H2O, and whose PaO2/FIO2 ratio was < or =160., Interventions: PaO2, PaCO2, pH, heart rate, blood pressure, and methemoglobin were recorded at baseline and after inhaling 1, 5, 10, and 20 ppm of nitric oxide. Peak expiratory flow rate and mean airway resistance were measured while subjects received 0 and 20 ppm of inhaled nitric oxide. Patients were followed up until extubation or death., Measurements and Main Results: Twenty-six patients (median age, 2.6 yrs [range, 1 mo-18.2 yrs]) were enrolled in the study. PaO2 increased (p< .001) and Pa(CO2) fell (p< .0001) from baseline with the administration of inhaled nitric oxide. There was no statistical difference among 1, 5, 10, and 20 ppm with regard to effects on oxygenation. Sixteen patients (62%) responded to inhaled nitric oxide with a > or =15% improvement in PaO2; 14 of these responses occurred at a dose of 1 or 5 ppm. Response to inhaled nitric oxide was not associated with age, length of intubation, presence of primary lung disease, chest radiograph, or illness severity. Among patients weighing < or =20 kg, responders showed a greater fall in mean airway resistance (p < .05) than nonresponders. Mortality was not influenced by prolonged inhaled nitric oxide when analyzed by intention to treat. Patients receiving prolonged inhaled nitric oxide at doses of < or =20 ppm maintained methemoglobin levels of <3.0% and circuit concentrations of NO2 of <1 ppm., Conclusions: Inhaled nitric oxide at doses of < or =5 ppm improves the oxygenation and (to a lesser extent) ventilation of most children with acute, hypoxic respiratory failure. The unpredictable response of patients necessitates individualized dosing of inhaled nitric oxide, starting at concentrations of < or =1 ppm. Inhaled nitric oxide at < or =20 ppm may exert a small salutary effect on bronchial tone. The benefits of prolonged inhaled nitric oxide remain unknown.

This study examined the distribution of two antifungal proteins, ribosome-inactivating protein (RIP) and zeamatin, in maize kernel tissues. Proteins were extracted from endosperm (including aleurone layer) and embryo tissues of imbibed maize kernels. Western blot analyses revealed that RIP-like protein was present at higher levels in endosperm than in embryo tissues, whereas zeamatin-like protein was more concentrated in embryo tissues than in endosperm tissues. However, there were three protein bands in the endosperm and two bands in the embryo that reacted to anti-RIP antibody in Western blot analyses. Tissue prints were conducted to localize the antifungal proteins. Imbibed kernels were cut longitudinally and transversely and blotted onto nitrocellulose membranes. Using antibodies against maize RIP and zeamatin, RIP was found primarily in the aleurone layer of the endosperm and glandular layer of scutellum, whereas zeamatin was located mainly in the kernel embryo. These results provide insight into the potential functions of these antifungal proteins, especially since the presence of RIP and zeamatin within maize kernels uniquely protects kernels from pathogens.

Many of the lepidopterous insects which attack sweet corn, Zea mays L., are susceptible to insecticidal proteins produced by Bacillus thuringiensis ssp. kurstaki (Berliner) (Btk). Transgenic sweet corn expressing a synthetic cry gene for production of a Btk-insecticidal protein may provide a more environmentally acceptable means of sweet corn production. Eight transgenic sweet corn hybrids containing a synthetic gene for CryIA(b) protein production (BT11 event) were evaluated for resistance to the corn earworm, Helicoverpa zea (Boddie), and fall armyworm, Spodoptera frugiperda (J. E. Smith). Laboratory tests revealed that all Btk sweet corn hybrids were highly resistant to leaf and silk feeding by neonate 3 and 6 d old corn earworm larvae. Ear damage in the field to the Btk sweet corn hybrids caused by corn earworm was negligible. All Btk sweet corn hybrids, except Btk 95-0901, were moderately resistant to leaf and silk feeding by the fall armyworm. Survival and weight gain were reduced when neonates were fed excised whorl leaves of the Btk plants. Weight gain, but not survival, was reduced when 3- and 6-d-old fall armyworm larvae were fed excised whorl leaves of the Btk plants. Btk sweet corn hybrids appear to be ideal candidates for use in integrated pest management (IPM) programs for both the fresh and processing sweet corn markets, and their use should drastically reduce the quantity of insecticides currently used to control these pests in sweet corn. With appropriate cultural practices, it is highly unlikely that Btk sweet corn will contribute to the development of resistance to Btk proteins in these insects because of the high toxicity of the Cry proteins expressed in these sweet corn hybrids and the harvest of sweet corn ears from fields before larvae can complete development.

Unlabelled: Gallium-67 has been a controversial tumor-imaging agent in nuclear medicine for decades. This controversy centers on why tumors are variable in gallium-avidity, whether 67Ga uptake is a transferrin-independent or dependent process, and whether tumors and normal tissues differ in mechanism of uptake. If the factors that control uptake of 67Ga were understood better, then efforts to improve oncologic imaging with 67Ga by increasing the tumor activity, or by decreasing the background, may be warranted., Methods: Conventional systems for evaluating the mechanism and control of 67Ga uptake have significant limitations. We have endeavored to circumvent these by developing a pair of transfected cell lines. One cell line has no transferrin receptor. In the other, the human transferrin receptor has been restored by transfection and is over-expressed constitutively, without the necessity to manipulate factors such as cell growth or iron content. The uptake of 67Ga, both as a citrate salt and as a gallium-transferrin complex, was examined in these pairs of cells in vitro. The effect of calcium and of soluble (ionic) iron concentration on 67Ga uptake also was determined. Tumors were grown as explants of these cells in nude mice and comparisons of uptake of 67Ga by these tumors in vivo were made., Results: The in vivo uptake of 67Ga is significantly increased in tumors in which the transferrin receptor is overexpressed, compared to those without a functional transferrin receptor. However, a notable amount of accumulation of 67Ga also occurs, both in vitro and in vivo, by a transferrin-independent route. In vitro experiments demonstrate that the uptake of 67Ga by the transferrin-independent route can be enhanced further to levels that equal or exceed those achieved by the transferrin-dependent route by increasing the content of calcium or iron salts in the incubation medium., Conclusion: Significant transferrin-independent uptake of 67Ga occurs both in vitro and in vivo. This uptake can be stimulated further in vitro, suggesting that in vivo enhancement might also be possible to enhance the utility of the radiometal for tumor imaging.

Somatometric parameters, renal size, and systolic blood pressure (SBP) were studied in 406 patients referred to pediatric nephrology and urology clinics. These patients included 269 females (66%), 67 African Americans (17%), and 87 patients with essential hypertension (21%). Z scores for the study population were comparable to published standards for height, kidney length, and SBP. Weight and body mass index scores were significantly greater than predicted from the standards, especially in the subset of patients with essential hypertension. Age, height, weight, body mass index, kidney length, and SBP all correlated with one another; however, on multiple regression analysis of SBP with the other five independent variables, only weight proved to have a significant correlation. Furthermore, the relationship of kidney length with SBP was positive and hypertensive patients had greater kidney size than published standards. These data do not support reduced kidney size in the population with essential hypertension, nor is there support for a convincing correlation between kidney length and SBP in the general pediatric population. Body weight correlates best with blood pressure. These findings warrant further study in a less-select population. Prevention and treatment of obesity may thus be of prime importance in addressing hypertension in children.

New methods are described that should facilitate high-resolution (5-10 A) image reconstructions from low-dose, low-contrast electron micrographs of frozen-hydrated specimens and processing of large, digital images produced by new imaging devices and modern electron microscopes. Existing techniques for automatic selection of images of individual biological macromolecules from electron micrographs are inefficient or unreliable. We describe the Crosspoint method (CP), which produces good quality solutions with relatively small miss rates and few false hits, and an extension of this method along with a procedure for refining its solution. Two algorithms for processing large images, one based on image subsampling, the other on image decomposition, are described. A large image is first compressed (e.g., by subsampling) and the CP method is applied to the compressed image to produce an initial solution. The information gathered at this stage is used to cut the original image into subimages and then to refine the particle coordinates in each subimage. An interactive environment for experimenting with particle identification methods is described., (Copyright 1997 Academic Press.)

The invasion of peanut (Arachis hypogaea L.) pods and seeds by aflatoxin-forming species of Aspergillus is linked to injury by the lesser cornstalk borer and frequently causes a severe reduction in crop quality. The lesser cornstalk borer is susceptible to the lepidopteran-active Bacillus thuringiensis insecticidal crystal protein. We have introduced a codon-modified Bacillus thuringiensis cryIA(c) gene into peanut using microprojectile bombardment. The toxin-coding region of a Bt cryIA(c) gene was reconstructed for expression in plants and the resulting 3.4 kb gene cassette (promoter: 1.8 kb coding: 3') was directly cloned into the BglII site of plant transformation vectors. The vectors contained the hph gene, conferring resistance to the antibiotic hygromycin. Somatic embryos initiated from immature peanut cotyledons of two cultivars were used as the target for bombardment. DNA from hygromycin-resistant embryogenic cell lines, regenerated plants, and a progeny plant showed the presence and integration of hph and Bt genes by PCR and/or Southern blot analyses. ELISA immunoassay of the CryIA(c) protein from the hygromycin-selected plants showed the expression of CryIA(c) protein up to 0.18% of total soluble protein. Insect feeding bioassay of transformed plants indicated various levels of resistance to the lesser cornstalk borer, from complete larval mortality to a 66% reduction in larval weight. A negative correlation between percent survival or larval weight and the amount of Bt CryIA(c) protein was recorded indicating in general that the higher the protein level the lower the survival or larval weight of the insect. Based on leaf bioassay, transformation of peanut with vectors containing the Bt cryIA(c) gene may be effective in protecting the peanut plants from damage by lepidopteran insect larvae of lesser cornstalk borer.

Electron-density averaging, fast Fourier synthesis and fast Fourier analysis programs have been adapted for parallel-computing systems. These have been linked to perform iterative phase improvement and extension utilizing non-crystallographic symmetry and solvent flattening. Various strategies for parallel algorithms have been tested on a variety of computers as a function of the number of computer nodes. Some experimental timing results are discussed.

The mechanism by which 67Ga accumulates in tumors is controversial. The most popular theory is that 67Ga binds to transferrin and gains access to cells by the transferrin receptor. However, substantial evidence suggests that uptake of 67Ga may not be universally mediated by transferrin in tumors. To determine whether transferrin is required for uptake of 67Ga in vivo, we compared the uptake of 67Ga by two types of implanted tumors and by normal tissues in normal and severely hypotransferrinemic strains of Balb/C mice. One type of tumor was strongly gallium-avid in normal mice; the other was not. Uptake of 67Ga by normal soft tissues was markedly less in hypotransferrinemic than in normal mice. Uptake of 67Ga by bone was equivalent in the two types of mice. For the more gallium-avid tumor, uptake of 67Ga was similar and the ratio of tumor-to-background activity was substantially higher in the hypotransferrinemic than in the normal mice. For the less gallium-avid tumor, uptake was significantly less in hypotransferrinemic than in normal mice. These data suggest that uptake of 67Ga by bone and by some tumors may be a transferrin-independent process.

Evidence has been presented both that metallothionein does and does not produce resistance to cisplatin. The metallothionein-enriched cells described in most previous studies have been selected for resistance to heavy metals, such as cadmium, or have been maintained in a medium enriched for the metals. Exposure to toxic metals could alter the cells in many ways. This report addresses the effect of metallothionein content alone, independent of exposure to metals, on cellular resistance to cisplatin. The toxicity of cisplatin was compared in NIH/3T3 cells that vary in their content of metallothionein as a consequence of transfection with a plasmid that results in the constitutive expression of metallothionein. The plasmid contains the bovine papillomavirus genome and the mouse metallothionein-I gene; it is driven by a glucose-regulated protein of 78 kD. Control cells were transfected with a similar plasmid in which the coding sequences for metallothionein were inverted and separated from the promoter, thereby abolishing expression. Expression of metallothionein required neither selection nor maintenance of cells in the presence of heavy metals. Despite large differences between the two types of cells in their cellular content of metallothionein and in their resistance to the toxicity of cadmium, no differences in resistance to cisplatin were observed.

L cells were enriched about 100-fold for catalase by the transcription of the transfected cDNA for human catalase. In spite of substantial enrichment for catalase, the transfected cells (LFN7C/B3) were not resistant to the cytotoxicity of a variety of agents which reduce dioxygen, including paraquat, menadione, adriamycin, phenazine methosulfate, and bleomycin. Instead, they were more sensitive to paraquat, bleomycin, and adriamycin than the untransfected cells from which they derived. Desferrioxamine afforded the cells enriched for catalase modest protection from the toxicities of bleomycin, paraquat, or adriamycin, suggesting that enrichment for iron as a consequence of the increased cellular content of catalase accounted for some of the increased sensitivities. The increased sensitivity of the LFN7C/B3 cells to bleomycin and paraquat was also attributed to the ability of catalase in cells to prevent the drug-induced consumption of O2; by capturing H2O2 before it can escape the cell and converting it to O2, catalase can maintain the concentration of O2 either for repeated rounds of chemical reduction or for direct interaction with the toxin. As predicted by this formulation, the toxicities of paraquat and of bleomycin were increased in an atmosphere of 40% dioxygen in comparison with an atmosphere of 20% dioxygen.

Dihydrofolate reductase-minus mutants of Chinese hamster ovary cells were depleted of glutathione peroxidase by transcription of the transfected bovine cDNA in inverted orientation upstream from the cDNA for dihydrofolate reductase to engender a bicistronic mRNA. In a clone of cells selected for expression of dihydrofolate reductase by the ability to grow in nucleoside-free medium the activity of glutathione peroxidase was reduced to 20% of the activity in the untransfected parental line of cells (DG44). The cells depleted of glutathione peroxidase were more sensitive to the toxicities of paraquat and adriamycin than the untransfected parental cells from which they derived but not more sensitive to bleomycin, menadione, or phenazine methosulfate. That the mildly increased sensitivity to paraquat and adriamycin was the consequence of the diminished cellular content of glutathione peroxidase was confirmed by the increase in sensitivity of untransfected cells after treatment with buthionine sulfoximine, an agent which depletes cells of glutathione. These and other data strongly suggest that the enzymic action of glutathione peroxidase protects cells from the toxicity of paraquat and adriamycin. The toxin which these agents engender is likely to be hydrogen peroxide or another hydroperoxide upon which glutathione peroxidase acts.

Reexamination of serum from a child thought to have died of ethylene glycol poisoning showed that the child had methylmalonic acidemia. The gas chromatographic peak identified as ethylene glycol by a clinical laboratory was actually due to propionic acid. Proof of a metabolic basis for the child's symptoms eventually exonerated his mother of the charge of murder.

To determine the relationship between cellular uptake of cadmium and content of metallothionein, we measured uptake of 109Cd in cells that differed in content of metallothionein (MT). MT cells were derived from NIH/3T3 cells by transfection with a plasmid containing the genome of bovine papilloma virus and the mouse metallothionein-I gene, driven by the promotor for the glucose-regulated protein of 78 kDa. Control cells were similarly transfected with bovine papilloma virus-based plasmids with the gene for metallothionein inverted and thus separated from the promoter (TM), or deleted, along with the promoter (BPA). The number of copies of bovine papilloma virus-based plasmids was similar in MT, TM, and BPA cells, approximately 100 per cell. MT cells were more than 10 times more resistant to the lethal effect of cadmium than were the control cells. Synthesis of metallothionein was 15-fold greater in the MT cells than in the TM or BPA cells. The uptake of 109Cd by the cells enriched in metallothionein was 4-fold less than by the control cells. These data suggest that an increased content of metallothionein may protect some cells from the toxic effects of cadmium, in part, by diminishing uptake of the metal.

Thirteen children (ages 9.2 +/- 3.7 years, mean +/- SD) received intravenous doses of labetalol, an alpha 1- and beta-adrenergic blocker, on 15 separate occasions for treatment of hypertension. In 12 of 15 episodes an initial dose of 0.55 +/- 0.34 mg/kg was given; in all 15 a continuous infusion of 0.78 +/- 0.39 mg/kg per hour was utilized for 67.3 +/- 57.1 hours. A significant decrease in systemic blood pressure occurred in all episodes (143/99.1 +/- 17.7/11.1 vs 115.6/72.4 +/- 7.7/9.5; p less than 0.01). A clinically unimportant yet statistically significant decrease in heart rate occurred during labetalol infusion (116.3 +/- 19.8 vs 107.8 +/- 11 beats/min; p less than 0.01). The episodes in children with creatinine clearances greater than 50 (n = 6) were compared with those with creatine clearances less than 20 ml/min per 1.73 m2 (n = 9); similar doses of labetalol were required for control of blood pressure. We conclude that infusion of labetalol is effective for control of blood pressure in children with hypertension, regardless of renal function.

Transhiatal esophagectomy with primary anastomosis to the stomach (gastric pull-up) is an attractive surgical alternative to colic interposition in patients with cancer of the esophagus and hypopharynx. However, the lack of intrinsic gastric peristalsis and complaints by patients ol postprandial regurgitation prompted us to measure the effect of body posture on the rates of gastric emptying in these patients. The rates of solid and liquid gastric emptying were measured in 14 patients who had undergone gastric interposition for esophageal and hypopharyngeal carcinoma. Rates of emptying were measured in both the supine and upright position using a dualisotope radiolabeling technique. In these patients, the rate of gastric emptying of both solids and liquids was significantly slower in the supine position than in the upright position. Emptying in supine patients was also prolonged when compared with supine normal volunteers. Conversely, the upright rate of solid and liquid emptying in the patients was accelerated when compared with published values for upright normal volunteers. We conclude that gastric emptying after gastric interposition is dependent on upright posture after meals., (Copyright © 1991 The Society of Thoracic Surgeons. Published by Elsevier Inc. All rights reserved.)

Paraquat-resistant HeLa cells were selected and characterized to determine the mechanism(s) of the toxic action of paraquat for cultured mammalian cells. From HeLa cells already selected for resistance to the toxicity of 40 microM paraquat more resistant cells were selected in 90 microM paraquat. In these more resistant cells (PQRHM90) one adaptation which occurred at the lesser concentration of paraquat, magnification of the cellular content of both the CuZn- and the Mn-containing superoxide dismutases, did not increase further. Instead, the cellular content of catalase and of glutathione peroxidase increased. The increased cellular content of glutathione peroxidase appeared more likely to have produced increased resistance to paraquat than did the augmented content of catalase because, after cultivation in the absence of paraquat, the cells retained both resistance to 90 microM paraquat and the increased content of glutathione peroxidase, while the content of catalase declined. The cellular content of reduced glutathione in the PQRHM90 cells grown in paraquat was diminished in comparison with that in the cells from which they derived. The data suggest that the PQRHM90 cells resisted the toxicity of paraquat by increasing the cellular activity of glutathione peroxidase as a means of detoxifying the H2O2 produced by paraquat.

Sodium polystyrene sulfonate (SPSS) is commonly administered for the acute and chronic treatment of hyperkalemia. Its oral intake is complicated by poor compliance due to multifaceted reasons. We therefore analyzed a method of reducing potassium (K) in formula by pretreatment with SPSS. If effective, this would bypass complications of enterally administered SPSS and provide low-K formula. Thirteen formulas and nutritional supplements were pretreated with SPSS to determine if one could bind K and provide formulas with decreased K contents. Using an SPSS concentration of 1 g/l mEq K in the formula, 62 +/- 2.6% (P less than 0.01, mean +/- SEM) of the K was removed in 30 min, while the sodium (Na) concentration was increased by 234 +/- 37% (P less than 0.01). Analysis suggests that the disproportionate increase in Na is due to exchange for calcium (Ca) and magnesium (Mg), interaction with proteins, and Na suspended with SPSS in the formula. Thus, SPSS pretreatment of formula is an effective method of making low-K formula, but the increase in Na exceeds the K reduction. Attention to possible complications of increased Na intake as well as decreased Ca and Mg intake is warranted.

Ultrafiltration (UF) using dialysate volumes of less than 20 ml/kg was assessed in 12 critically ill infants and children with hypervolemia as the primary indication for dialysis. Ten of the 12 required mechanical ventilation (O2 requirements 45-100%). Seven, all infants, received at least one pressor intravenously in attempts to maintain systolic blood pressure of 60. Age was 22 +/- 11.5 mos. (mean +/- SE) with weight of 11.1 +/- 4.3 kg (range 2.4-50 kg). Dialysis prescription was as follows: Volume of 10.1 +/- 1.3 ml/kg/exchange; dwell time 30-45 minutes; drain time 15-20 minutes. Glucose concentration ranged from 1.5-4.25%. Drained dialysate volume expressed as a percent of volume instilled was 133.5 +/- 3.4%. When calculated for body weight and time, UF was 3.0 +/- 0.3 ml/kg/hour. UF was not significantly different in patients requiring pressors versus those not requiring blood pressure support. Although minor ventilatory changes were required, significant deterioration of blood gases did not occur during the first 3-6 hours of PD, nor did significant changes in blood pressure develop. Hyperkalemia, present in 1 patient prior to PD, resolved using this regimen. Low volume PD was a viable alternative to hemodialysis or hemofiltration for control of hypervolemia in this small group of very ill infants and children.