Strengthening the Chain of Survival: Cardiopulmonary Resuscitation Workshop for Caregivers of Children at Risk.

BACKGROUND: Parents and caregivers should receive training regarding pediatric cardiopulmonary resuscitation (CPR) because this knowledge improves survival. We conducted a study as part of a Patient Safety Project to improve caregivers' CPR knowledge and skills. We also aimed to improve the quality of patient care. METHODS: We performed a prospective, longitudinal study in 2013-2014 in a pediatric hospital. We enrolled the caregivers of all patients admitted with a diagnosis of an acute life-threatening event, apnea, or choking. We provided a 45-minute CPR workshop for parents at discharge and evaluated the results using a test before, immediately after, and at 1 and 3 months after the workshop. Participants also completed an evaluation survey about the CPR workshop. RESULTS: We admitted 62 patients [median age, 1 mo (0.5-2 mo)]. We provided 62 pediatric CPR workshops to 106 enrolled relatives. The median score was 5 (CI, 3-6) out of 10 at baseline, which increased to 8 (CI, 7-10) immediately after the workshop (P < 0.01). After 1 and 3 months, the median score was 8 (CI, 6-9; P < 0.01). The severity of the acute life-threatening event episode correlated with a better score (P = 0.02). The utility of the workshop scored 9.9 out of 10. CONCLUSIONS: This CPR workshop significantly increased CPR knowledge and confidence, and this was maintained up to 3 months post-training. Caregiver satisfaction was high.

The Complexity of Antibody Responses Elicited against the Respiratory Syncytial Virus Glycoproteins in Hospitalized Children Younger than 2 Years.

The influence of age and maternal antibodies on the antibody responses to human respiratory syncytial virus (hRSV) glycoproteins in very young children has been a matter of controversy. Both, immaturity of the immune system at very early age and suppression of the host immune response by high level of maternal antibodies have been claimed to limit the host antibody response to virus infection and to jeopardize the use of hRSV vaccines under development in that age group. Hence, the antibody responses to the two major hRSV glycoproteins (F and G) were evaluated in children younger than 2 years, hospitalized with laboratory confirmed hRSV bronchiolitis. A strong negative correlation was found between the titre of circulating ELISA antibodies directed against either prefusion or postfusion F in the acute phase, but not age, and their fold change at convalescence. These changes correlated also with the level of circulating neutralizing antibodies in sera. As reported in adults, most neutralizing antibodies in a subset of tested sera could not be depleted with postfusion F, suggesting that they were mostly directed against prefusion-specific epitopes. In contrast, a weak negative association was found for group-specific anti-G antibodies in the acute phase and their fold change at convalescence only after correcting for the antigenic group of the infecting virus. In addition, large discrepancies were observed in some individuals between the antibody responses specific for F and G glycoproteins. These results illustrate the complexity of the anti-hRSV antibody responses in children experiencing a primary severe infection and the influence of preexisting maternal antibodies on the host response, factors that should influence hRSV serological studies as well as vaccine development.

Halogen mediated voltammetric oxidation of biological thiols and disulfides.

The electrochemical generation of the halides, bromine and iodine, in the presence of biologically relevant organosulfur is demonstrated to result in an analytically useful response. In the case of the iodide/iodine redox couple only the thiol causes an increase in the electrochemical oxidative peak current. Conversely, the formed bromine may catalytically oxidise both thiols and disulfides. Hence, the differing reactivities of the halide ions readily allow discrimination between the closely related thiol and disulphide species. For all of the organosulfur species investigated (glutathione, cysteine and homocysteine) micromolar limits of detection are attainable. In the case of the bromine mediated oxidation this sensitivity at least partially arises from the large catalytic amplification, such that, for each disulphide molecule up to ten electrons may be transferred. Ultimately this bromine oxidation results in the formation of the sulfonate species. For the iodine mediated oxidation of the thiols the oxidation proceeds no further than to the formation of the associated disulfide.

Catalase-like activity of human methemoglobin: a kinetic and mechanistic study.

Hydrogen peroxide triggers a redox cycle between methemoglobin and ferrylhemoglobin, leading to protein inactivation and oxygen evolution. In the present paper, the catalase-like oxygen production by human methemoglobin in the presence of H(2)O(2) was kinetically characterized with a Clark-type electrode. Progress curves showed a pseudo-steady state in the first minutes of the reaction, while double-reciprocal plots were upwardly concave, indicating positive co-operativity dependent upon protein concentration, which is a very unusual kinetic behavior. Addition of superoxide radical scavengers slightly increased activity, suggesting that most oxygen was produced biocatalytically. By considering all the experimental data obtained, a possible mechanism was proposed, including: (a) competition between the one-electron and the two-electron reductions of the oxoferryl free radical species of hemoglobin, giving rise to ferrylhemoglobin and methemoglobin, respectively; (b) competition between the superoxide-dependent inactivation of the protein and its reduction back to the met state. Computer simulations of that model have been performed by numerically integrating the differential equations set describing the mechanism, which was seen to yield predictions of the kinetic parameters variation consistently with the kinetic behavior experimentally observed. We suggest that the catalase-like activity of methemoglobin must predominantly be a biocatalytic reaction that protects the protein against H(2)O(2)-induced suicide inactivation.

Mechanism of acetaminophen oxidation by the peroxidase-like activity of methemoglobin.

Oxidation of acetaminophen by human methemoglobin in the presence of H(2)O(2) has been kinetically studied in the present paper. The drug showed a protective effect against the H(2)O(2)-induced irreversible inactivation of the protein, thus indicating the competition among both ligands, H(2)O(2) and acetaminophen for the protein. The stoichiometry of the reaction is variable and depends on relative initial concentrations of H(2)O(2) and the drug owing to their competitive behavior. In addition and unexpectedly, the protein exhibits non Michaelian kinetics against both acetaminophen and H(2)O(2) under steady-state conditions and shows negative co-operativity with Hill coefficients in the 0.3-0.7 range. Therefore, these data were compared to those obtained with myoglobin under similar experimental conditions, and the same results were observed. This led us to propose a mechanism for the peroxidase-like activity of hemoglobin, which accounts for the experimental results obtained herein. The steady-state rate equation for this mechanism has been obtained and is also consistent with the experimental data, thus indicating the goodness of the model proposed herein. The results presented in this work provide new insights into the oxidation mechanism of acetaminophen.

Computer simulation of the dynamic behavior of the glutathione-ascorbate redox cycle in chloroplasts.

The glutathione-ascorbate redox pathway in chloroplasts is a complex network of spontaneous, photochemical, and enzymatic reactions for detoxifying hydrogen peroxide. This article presents a comprehensive sensitivity analysis of the system. A model has been constructed to simulate oxidative stress conditions, enabling steady-state concentrations of the metabolites involved in the pathway and photochemical and enzymatic fluxes to be calculated. The model includes an electron source whose flux is distributed among three competitive routes (photogeneration of O2-, photoreduction of NADP+ to NADPH, and photoreduction of monodehydroascorbate to ascorbate) and that allows the simulation of variations in NADPH concentration with time. Each enzyme considered is introduced in the model, taking into account its particular catalytic mechanism, including the inactivation of ascorbate peroxidase in the presence of low-ascorbate concentrations. Computer simulations pointed to the great sensitivity of the system to the ratio among fluxes corresponding to ascorbate and NADPH photoproduction and NADPH consumption by the Calvin cycle. Under oxidative stress conditions, the model shows a sequential depletion of antioxidant power in chloroplasts in the order NADPH, glutathione, ascorbate and their recovery in the reverse order. Decreasing levels of glutathione reductase, ascorbate peroxidase, and superoxide dismutase led to the irreversible photoinactivation of ascorbate peroxidase and the subsequent increase in hydrogen peroxide concentration, preceded by a maximum in dehydroascorbate reductase activity.