Postoperative pain assessment in children: a pilot study of the usefulness of the analgesia nociception index.

BACKGROUND: The ability to perform objective pain assessment is very important in paediatric patients. The goal of this study was to investigate the relationship between the analgesia nociception index (ANI), which is based on the heart rate variability, and objective measurements of pain intensity in young or cognitively impaired children, after surgical or imaging procedures (control group) under general anaesthesia. METHODS: On arrival in the recovery room and subsequently at 5-10 min intervals, the level of pain was rated using the FLACC pain scale (0-10). The ANI values (0-100; 0 indicating the worst pain) were recorded simultaneously. The area under the receiver operating characteristic curve (AUC) and grey zone approach were used to evaluate the performance of the ANI to detect patients with FLACC >4. Instantaneous ANI values were compared with ANI values averaged over 256 s periods of time. RESULTS: All children in the surgical group (n=32) developed moderate-to-severe pain (FLACC >4). Children in the control group (n=30) exhibited minimal pain. Instantaneous ANI values were lower in children of the surgical group than in the control group [52 (sd16) vs 69 (16), P<0.001]. The AUC for the 256 s ANI recording period [0.94 (95% confidence interval 0.85-0.99)] was significantly higher than for instantaneous ANI (P<0.05). When measured for a period of 256 s, an ANI cut-off value of 56 (grey zone [58-60]) was most predictive of a FLACC ≥4. CONCLUSIONS: The ANI may provide an objective measurement of acute postoperative pain, which is correlated with that measured on a FLACC scale in young or cognitively impaired children.

A socio-ecological approach promoting physical activity and limiting sedentary behavior in adolescence showed weight benefits maintained 2.5 years after intervention cessation.

BACKGROUND: Obesity in youth remains a major public health issue. Yet no effective long-term preventive strategy exists. We previously showed that a school-based socio-ecological approach targeting behavior and social/environmental influences on physical activity (PA) prevented 4-year excessive weight gain in 12-year olds. In this study, we investigated if this efficacy persists 30 months after intervention cessation. METHODS AND FINDINGS: The program targeted students, family, school and the living environment to promote/support PA and prevent sedentary behavior (SB). A total of 732 students from eight randomized middle schools completed the 4-year trial. At the 30-month post-trial follow-up, body mass index (BMI), fat mass index (FMI), leisure PA (LPA), home/school/workplace active commuting, TV/video time (TVT), and attitudes toward PA were measured in 531 adolescents. The beneficial effects of the intervention on the excess BMI increase (+0.01 vs +0.34 kg m(-2) in the intervention and control groups, respectively) and on the overweight incidence in initially non-overweight students (4.3% vs 8.6%; odds ratio=0.48 (95% confidence interval: 0.23-1.01)) were maintained at the post-trial follow-up. LPA was not maintained at the level achieved during the trial. However, we still observed a prevention of the age-related decrease of the adolescents' percentage reporting regular LPA (-14.4% vs -26.5%) and a higher intention to exercise in the intervention group. The intervention promoted lower TVT (-14.0 vs +13.6 min per day) and higher active commuting changes (+11.7% vs -4.8%). Trends in higher BMI reduction in students with high initial TVT and in the least wealthy group were noted. TVT changes throughout the follow-up predicted excess BMI and FMI changes. CONCLUSIONS: Long-term multilevel approach targeting PA and SB prevents excessive weight gain up to 30 months after intervention cessation. The efficacy may be higher in the most sedentary and least wealthy adolescents. Healthy PA-related behavior inducing long-lasting weight effects can be promoted in youth providing that an ecological approach is introduced in the prevention strategy.

Spatial accessibility to physical activity facilities and to food outlets and overweight in French youth.

OBJECTIVE: Some characteristics of the built environment have been associated with obesity in youth. Our aim was to determine whether individual and environmental socio-economic characteristics modulate the relation between youth overweight and spatial accessibility to physical activity (PA) facilities and to food outlets. DESIGN: Cross-sectional study. SUBJECTS: 3293 students, aged 12 ± 0.6 years, randomly selected from eastern France middle schools. MEASUREMENTS AND METHODS: Using geographical information systems (GIS), spatial accessibility to PA facilities (urban and nature) was assessed using the distance to PA facilities at the municipality level; spatial accessibility to food outlets (general food outlets, bakeries and fast-food outlets) was calculated at individual level using the student home address and the food outlets addresses. Relations of weight status with spatial accessibility to PA facilities and to food outlets were analysed using mixed logistic models, testing potential direct and interaction effects of individual and environmental socio-economic characteristics. RESULTS: Individual socio-economic status modulated the relation between spatial accessibility to PA facilities and to general food outlets and overweight. The likelihood of being overweight was higher when spatial accessibility to urban PA facilities and to general food outlets was low, but in children of blue-collar-workers only. The odds ratio (OR) (95% confidence interval) for being overweight of blue-collar-workers children compared with non-blue-collar-workers children was 1.76 (1.25-2.49) when spatial accessibility to urban PA facilities was low. This OR was 1.86 (1.20-2.86) when spatial accessibility to general food outlets was low. There was no significant relationship of overweight with either nature PA facilities or other food outlets (bakeries and fast-food outlets). CONCLUSION: These results indicate that disparities in spatial accessibility to PA facilities and to general food outlets may amplify the risk of overweight in socio-economically disadvantaged youth. These data should be relevant for influencing health policies and urban planning at both a national and local level.

Perceived barriers and facilitators for general practitioner-patient communication in palliative care: a systematic review.

While effective general practitioner (GP)-patient communication is required for the provision of good palliative care, barriers and facilitators for this communication are largely unknown. We aimed to identify barriers and facilitators for GP-patient communication in palliative care. In a systematic review seven computerized databases were searched to find empirical studies on GP-patient communication in palliative care. Fifteen qualitative studies and seven quantitative questionnaire studies were included. The main perceived barriers were GPs' lack of availability, and patients' and GPs' ambivalence to discuss 'bad prognosis'. Main perceived facilitators were GPs being available, initiating discussion about several end-of-life issues and anticipating various scenarios. Lack of availability and failure to discuss former mistakes appear to be blind spots of GPs. GPs should be more forthcoming to initiate discussions with palliative care patients about prognosis and end-of-life issues. Empirical studies are needed to investigate the effectiveness of the perceived barriers and facilitators.

Seroprevalence of West Nile virus infection in solid organ transplant recipients.

BACKGROUND: Of people infected with mosquito-borne West Nile virus (WNV), <1% develop neuroinvasive disease (NID). Population studies suggest that people older than 65 years may be at higher risk for neurologic symptoms. It has been suggested that solid organ transplant (SOT) recipients are also at higher risk for WNV NID, but definitive serologic and epidemiologic data are lacking. METHODS: A serologic screening survey, using a US Food & Drug Administration-approved enzyme-linked immunosorbant assay to detect WNV immunoglobulin-G (IgG) antibody responses in cohorts of SOT recipients and non-immunocompromised controls, was undertaken at a large Midwestern university organ transplant center in the aftermath of the summer 2003 WNV regional outbreak. Hemagglutination-inhibition testing was used to confirm WNV IgG-positive results and differentiate them from positive results caused by Saint Louis encephalitis virus, another flavivirus that is endemic in the Midwestern US. FINDINGS: The rate of WNV IgG-seropositive responses did not differ between SOT recipients and non-immunocompromised controls, and were 12% and 10%, respectively. Retrospective chart review showed no documented WNV NID in the seropositive SOT recipients, suggesting an incidence of WNV NID may be as low as 0.7% in this population. INTERPRETATION: Asymptomatic WNV infection is common among immunocompromised SOT patients, occurring as often as it does in non-immunocompromised controls. Our data indicated that severe WNV NID is less frequent in SOT patients, contrary to what has been suggested in other studies.

Synthesis of globin RNA in enucleated differentiating murine erythroleukemia cells.

In an earlier report (Volloch, V., 1986, Proc. Natl. Acad. Sci. USA., 83:1208-1212) we had presented evidence for the occurrence of the cytoplasmic synthesis of globin mRNA and of RNA complementary to globin mRNA which differed from DNA-dependent transcription by its insensitivity to actinomycin D. In this paper, we describe the use of enucleated differentiating mouse erythroleukemia cells to demonstrate directly the occurrence of cytoplasmic synthesis of both positive- and negative-strand globin RNA. For this purpose, we developed an enucleation procedure which yielded pure cytoplasts from differentiated mouse erythroleukemia cells in the absence of cytochalasin B and selectively permeabilized the cytoplasts to small molecules by treatment with dextran sulfate and saponin. The permeabilized cytoplasts incorporated [3H]dUTP into positive- and negative-strand globin RNA and experiments with mercurated nucleotide substrate suggested that this process involved de novo RNA synthesis rather than limited terminal nucleotide addition. Globin RNA synthesis required Mg++, was inhibited by Mn++, and was unaffected by the addition of Zn++. Studies of its response to inhibitors of DNA-dependent RNA synthesis showed that it differed from that process in its insensitivity to actinomycin D and alpha-amanitin, but that like many other macromolecular biosynthetic reactions it was inhibited by rifamycin AF/ABDP and aurintricarboxylic acid. These observations provide additional evidence for the occurrence of cytoplasmic RNA-dependent RNA synthesis in differentiated cells and show permeabilized enucleated cells to be a useful experimental system for further studies of the characteristics of that process.

Successful overweight prevention in adolescents by increasing physical activity: a 4-year randomized controlled intervention.

BACKGROUND: Population-based studies directed at promoting physical activity in youth have shown limited success in obesity prevention. OBJECTIVE: To assess whether an intervention integrating environmental changes to induce sustained changes in physical activity, prevents overweight in adolescents. DESIGN: Four-year randomized trial started in 2002 in eight middle schools of Eastern France. The intervention, randomized at school level, was designed to promote physical activity by changing attitudes through debates and attractive activities, and by providing social support and environmental changes encouraging physical activity. SUBJECTS: Nine hundred and fifty four 12-year-old six-graders. MEASUREMENTS: Body mass index (BMI), body composition, physical activity by questionnaire, plasma lipids and glucose, insulin resistance. RESULTS: Intervention students had a lower increase in BMI (P=0.01) and age- and gender-adjusted BMI (P<0.02) over time than controls. The differences across groups of the age- and gender-adjusted BMI changes (95% confidence interval (CI)) were -0.29 (-0.51; -0.07) kg/m2 at 3 years, -0.25 (-0.51; 0.01) kg/m2 at 4 years. An interaction with baseline weight status was noted. The intervention had a significant effect throughout the study in initially non-overweight adolescents (-0.36 (-0.60;-0.11) kg/m2 for adjusted BMI at 4 years), corresponding to a lower increase in fat mass index (P<0.001). In initially overweight adolescents, the differences observed across groups at 2 years (-0.40 (-0.94; 0.13) kg/m2 for adjusted BMI) did not persist over time. At 4 years, 4.2% of the initially non-overweight adolescents were overweight in the intervention schools, 9.8% in the controls (odds ratio=0.41 (0.22; 0.75); P<0.01). Independent of initial weight status, compared with controls, intervention adolescents had an increase in supervised physical activity (P<0.0001), a decrease of TV/video viewing (P<0.01) and an increase of high-density cholesterol concentrations (P<0.0001). CONCLUSION: Enhancing physical activity with a multilevel program prevents excessive weight gain in non-overweight adolescents. Our study provides evidence that prevention of obesity in youth is feasible.

Adiponectin is associated with lipid profile and insulin sensitivity in French adolescents.

AIM: The favourable relationship of adiponectin with the metabolic profile demonstrated in adults has been less studied in youths. The aim of this study was to examine cross-sectional and longitudinal associations between adiponectin and various metabolic risk factors in 12-year-old adolescents. METHODS: Subjects were participants in a randomized controlled study to promote physical activity (PA). Cross-sectional associations were assessed at entry in 2002 among 647 PA-exposed and control first-level students (49% male, 11.6+/-0.6 years of age). Longitudinal analyses involved 288 control students surveyed in 2002 and 2004. Baseline measurements included fasting serum adiponectin and anthropometric indices (body mass, waist size, body fat [BF] by bioimpedance), insulin concentration, homeostasis model assessment (HOMA), high-density lipoprotein (HDL) cholesterol, triglycerides (TG), soluble TNF-alpha receptor 1 (sTNF-alpha R1) and high-sensitivity C-reactive protein. Analyses were performed with generalized linear mixed-effects models, taking into account correlations among adolescents in the same school. RESULTS: Cross-sectionally, plasma adiponectin was inversely associated with obesity indices, especially waist size (P<10(-2)), HOMA (P<0.03), insulin (P<0.04), TG (P<10(-2)) and sTNF-alpha R1 (P<0.05), and positively related to HDL cholesterol (P<10(-4)), after adjusting for age, gender, sexual maturity, sports participation and adiposity when relevant. Longitudinally, a higher baseline adiponectin level was associated with a more favourable two-year change in TG (P<0.05), even after accounting for baseline TG, and two-year BF and insulin changes. CONCLUSION: The findings of this study suggest a favourable relationship between adiponectin and both metabolic profile and subsequent changes in TG level in young adolescents.

ICAPS: a multilevel program to improve physical activity in adolescents.

OBJECTIVES: ICAPS (Intervention Centred on Adolescents' Physical activity and Sedentary behaviour) is aimed at preventing excessive weight gain and cardiovascular risk in adolescents by promoting physical activity (PA) with an emphasis on recreational and daily-life PA, with a lifelong perspective. DESIGN: Randomized study designed to last for four years. Study cohort constituted of 954 first-level students (91% of eligible pupils), aged 11.7 +/- 0.6 y (mean +/- SD) from four pairs of schools randomly selected in eastern France, after sociogeographical stratification. In each pair, intervention status was randomised at school-level. The program, not limited to school settings, involves multiple partners with three objectives: 1) changing attitudes through debates and access to attractive activities during breaks and after-school hours, 2) encouraging social support, 3) providing environmental conditions that enable PA. Adapted times and places, open participation, emphasis on fun, meeting with others and absence of competitive aspects are used to reduce usual barriers to PA. Accessibility and safety are permanent concerns. RESULTS: Prevalence of overweight was 23.7%. High participation rates were attained (50% participated in at least one weekly activity). At six-month, the proportion of intervention adolescents not performing supervised PA out of academic PA was reduced by half (36% to 17% vs 42% to 42% in controls P < 10-4); the proportion of those spending > 3 h/day in sedentary occupations decreased (34% to 28% vs 27% to 36%; P < 10-4). CONCLUSION: These data demonstrate the feasibility of implementing a multilevel PA intervention program in adolescents. Six-month results document increased PA and decreased sedentary behaviour.

Signal amplification by rolling circle amplification on DNA microarrays.

While microarrays hold considerable promise in large-scale biology on account of their massively parallel analytical nature, there is a need for compatible signal amplification procedures to increase sensitivity without loss of multiplexing. Rolling circle amplification (RCA) is a molecular amplification method with the unique property of product localization. This report describes the application of RCA signal amplification for multiplexed, direct detection and quantitation of nucleic acid targets on planar glass and gel-coated microarrays. As few as 150 molecules bound to the surface of microarrays can be detected using RCA. Because of the linear kinetics of RCA, nucleic acid target molecules may be measured with a dynamic range of four orders of magnitude. Consequently, RCA is a promising technology for the direct measurement of nucleic acids on microarrays without the need for a potentially biasing preamplification step.

Intrinsic resistance to methotrexate in human soft tissue sarcoma cell lines.

A human fibrosarcoma cell line, HT-1080, and four new cell lines (HS-16, HS-28, HS-30, and HS-42) were established from untreated patients with mesenchymal chondrosarcoma, peripheral nerve sheath sarcoma, malignant hemangiopericytoma, and mixed mesodermal tumor, respectively, and were used for analysis of mechanisms of intrinsic resistance to methotrexate. All four new cell lines were resistant to methotrexate as determined by inhibition of thymidylate synthase in whole cells and by growth inhibition, as compared with HT-1080, a methotrexate sensitive cell line. Methotrexate uptake, level of dihydrofolate reductase, and inhibition of this enzyme by methotrexate in the four cell lines were comparable to HT-1080 cells. However, levels of long chain polyglutamates (glu3-5) of methotrexate achieved after a 24-h incubation with this drug were much lower in the four new cell lines as compared to the HT-1080 cell line (5- to 20-fold lower). The low levels of methotrexate polyglutamates formed is likely the major cause of intrinsic methotrexate resistance in these new sarcoma cell lines.

Dyspnoea: Pathophysiology and a clinical approach.

Dyspnoea, also known as shortness of breath or breathlessness, is a subjective awareness of the sensation of uncomfortable breathing. It may be of physiological, pathological or social origin. The pathophysiology of dyspnoea is complex, and involves the activation of several pathways that lead to increased work of breathing, stimulation of the receptors of the upper or lower airway, lung parenchyma, or chest wall, and excessive stimulation of the respiratory centre by central and peripheral chemoreceptors. Activation of these pathways is relayed to the central nervous system via respiratory muscle and vagal afferents, which are consequently interpreted by the individual in the context of the affective state, attention, and prior experience, resulting in the awareness of breathing. The clinical evaluation and approach to the management of dyspnoea are directed by the clinical presentation and underlying cause. The causes of dyspnoea are manifold, and include a spectrum of disorders, from benign to serious and life-threatening entities. The pathophysiology, aetiology, clinical presentation and management of dyspnoea are reviewed.

Combining nucleic acid amplification and detection.

Major recent advances in molecular amplification in the past year were initial validation of two new amplification technologies (rolling circle amplification and Invader), a significant increase in the number of molecular diagnostic assays, achievement of amplification directly on microarrays (by strand displacement amplification and rolling circle amplification), and description of two new read-out probes (Scorpions and nanoparticles).

Decreased polyglutamylation of methotrexate in acute lymphoblastic leukemia blasts in adults compared to children with this disease.

We compared blast cells from adult and pediatric patients with untreated acute lymphoblastic leukemia (ALL) (as separated groups of T-lineage cell and B-lineage cell ALL) to determine if methotrexate (MTX) polyglutamate formation in adult patients might be a contributing cause to the known difference in clinical outcome, since MTX is a key drug in chemotherapy regimens. Adult B-lineage cell ALL blasts and blasts from the patients with T-lineage cell ALL accumulated lower amounts of total MTX and polyglutamates, especially long-chain MTX polyglutamates (glu3-6) than pediatric B-lineage cell ALL blasts. In view of the importance of polyglutamylation of MTX as a determinant of cytotoxicity of this drug, decreased formation of MTX polyglutamates is likely a contributing cause to the lower cure rate in adult ALL and T-lineage cell ALL as compared to childhood B-lineage cell ALL.

Effects of ß2 agonists on post-thoracotomy pain incidence.

BACKGROUND: Pre-clinical research has shown ß2 -adrenoceptors to be essential for the antiallodynic action of antidepressant drugs in murine models of neuropathic pain and that sustained treatment with ß2 -agonists has an antiallodynic action. Here, we clinically investigated whether chronic ß2 -agonist treatments may influence the incidence of post-thoracotomy chronic pain, defined as pain that recurs or persists along a thoracotomy scar more than 2 months after surgery, either neuropathic or non-neuropathic. METHODS: We conducted an epidemiological study on patients operated by thoracotomy. Demographic data, medical history and treatments concomitant to the surgery were recorded at a follow-up visit. Information on perioperative treatments was collected from the anaesthesia records and confirmed by the patients. In patients with pain at the surgery level, post-thoracotomy chronic pain was assessed by clinical examination and numeric scale. Physical examination and DN4 questionnaire were used to discriminate neuropathic and non-neuropathic chronic pain at scar level. RESULTS: One hundred and eighty-nine patients were included. Eighty-one patients reported persisting thoracic pain, with neuropathic characteristics in 58 of them (30% of the 189 patients). The most common chronic drugs during the perioperative period were inhaled ß2 -agonists (28.6%). The chronic use of ß2 -agonists was an independent predictor of thoracic neuropathic pain (but not of non-neuropathic pain) and was associated with a five-fold decrease in the relative incidence of neuropathic pain [OR = 0.19 (0.06-0.45)]. CONCLUSIONS: These data suggest a possible influence of chronic ß2 -agonist treatments on neuropathic pain secondary to thoracotomy. This apparent preventive effect of ß2 -agonist treatments should warrant controlled clinical trials.

Comparison of the expression of a mutant dihydrofolate reductase under control of different internal promoters in retroviral vectors.

To determine the effect of different promoters on the expression of an altered dihydrofolate reductase (DHFR) gene conferring methotrexate (MTX) resistance in different cell types, double-copy retroviral vectors were constructed carrying a murine mutant DHFR under the control of five different promoters, i.e., human adenosine deaminase (ADA), simian virus 40 (SV40), thymidine kinase (TK), human beta-actin, and cytomegalovirus (CMV). Their expression was compared in NIH-3T3 cells, three human leukemia cell lines, and mouse bone marrow. The variant DHFR is readily expressed from these various promoters in retroviral vectors at a selectable level. In 3T3 cells, the DHFR constructs containing the SV40 promoter conferred the highest levels of resistance to MTX. In K562 and Raji cells, the construct with the TK promoter produced the highest level of resistance. However granulocyte-macrophage colony-forming unit (CFU-GM) colonies from mouse marrow were more resistant to MTX when infected with vectors containing the SV40 promoter and ADA promoter as compared to the other promoter constructs. These studies show that mouse fibroblast cell lines such as NIH-3T3 do not predict the effectiveness of retroviral-mediated gene transfer for marrow progenitor cells, and that the activity of retroviral vector-encoded promoters vary in an unpredictable manner from cell type to cell type. Possible implications for basic gene transfer studies and clinical applications are discussed.

The nucleotide sequence of the yeast ARG4 gene.

The complete nucleotide sequence of a 2296-bp DNA fragment containing the yeast (Saccharomyces cerevisiae) ARG4 gene has been determined. This gene specifies the synthesis of the arginine biosynthetic enzyme, argininosuccinate lyase (EC 4.3.2.1). The sequence contains one major open reading frame of 463 codons, giving a calculated Mr of 52 010 for the protein, in good agreement with the experimentally determined value of 53 000. The sequence upstream from the ARG4 gene shares structural features in common with other yeast genes subject to general amino acid control.

Transfection with a cDNA encoding a Ser31 or Ser34 mutant human dihydrofolate reductase into Chinese hamster ovary and mouse marrow progenitor cells confers methotrexate resistance.

Chinese hamster ovary (CHO) DHFR- cells were converted into the DHFR+ phenotype when they were transfected with a mammalian expression vector carrying human dihydrofolate reductase-encoding cDNAs (DHFR) containing a Ser31 or a Ser34 mutation. Furthermore, transfection of these mutants into wild-type CHO cells resulted in resistance to high levels of methotrexate (MTX), indicating that these human variants can act as dominant selectable markers. Southern blot analysis and polymerase chain reaction amplifications confirmed that the transfected plasmids were integrated into the CHO DNA. Gene copy number analysis revealed that both the Ser3 1 and the Ser3.4 mutants amplifiable when grown in increasing concentrations of MTX. Retrovirus-mediated gene transfer of the Ser31 mutant into mouse marrow progenitor cells also resulted in MTX-resistant CFU-GM (colony-forming unit-granulocyte macrophage) cells.

Effects of nucleoside analog incorporation on DNA binding to the DNA binding domain of the GATA-1 erythroid transcription factor.

We investigate here the effects of the incorporation of the nucleoside analogs araC (1-beta-D-arabinofuranosylcytosine) and ganciclovir (9-[(1,3-dihydroxy-2-propoxy)methyl] guanine) into the DNA binding recognition sequence for the GATA-1 erythroid transcription factor. A 10-fold decrease in binding affinity was observed for the ganciclovir-substituted DNA complex in comparison to an unmodified DNA of the same sequence composition. AraC substitution did not result in any changes in binding affinity. 1H-15N HSQC and NOESY NMR experiments revealed a number of chemical shift changes in both DNA and protein in the ganciclovir-modified DNA-protein complex when compared to the unmodified DNA-protein complex. These changes in chemical shift and binding affinity suggest a change in the binding mode of the complex when ganciclovir is incorporated into the GATA DNA binding site.

Accessibility of selenomethionine proteins by total chemical synthesis: structural studies of human herpesvirus-8 MIP-II.

The determination of high resolution three-dimensional structures by X-ray crystallography or nuclear magnetic resonance (NMR) is a time-consuming process. Here we describe an approach to circumvent the cloning and expression of a recombinant protein as well as screening for heavy atom derivatives. The selenomethionine-modified chemokine macrophage inflammatory protein-II (MIP-II) from human herpesvirus-8 has been produced by total chemical synthesis, crystallized, and characterized by NMR. The protein has a secondary structure typical of other chemokines and forms a monomer in solution. These results indicate that total chemical synthesis can be used to accelerate the determination of three-dimensional structures of new proteins identified in genome programs.