Evaluation of tick-borne encephalitis DNA vaccines in monkeys.

Tick-borne encephalitis is usually caused by infection with one of two flaviviruses: Russian spring summer encephalitis virus (RSSEV) or Central European encephalitis virus (CEEV). We previously demonstrated that gene gun inoculation of mice with naked DNA vaccines expressing the prM and E genes of these viruses resulted in long-lived homologous and heterologous protective immunity (Schmaljohn et al., 1997). To further evaluate these vaccines, we inoculated rhesus macaques by gene gun with the RSSEV or CEEV vaccines or with both DNA vaccines and compared resulting antibody titers with those obtained by vaccination with a commercial, formalin-inactivated vaccine administered at the human dose. Vaccinations were given at days 0, 30, and 70. All of the vaccines elicited antibodies detected by ELISA and by plaque-reduction neutralization tests. The neutralizing antibody responses persisted for at least 15 weeks after the final vaccination. Because monkeys are not uniformly susceptible to tick-borne encephalitis, the protective properties of the vaccines were assessed by passive transfer of monkey sera to mice and subsequent challenge of the mice with RSSEV or CEEV. One hour after transfer, mice that received 50 microl of sera from monkeys vaccinated with both DNA vaccines had circulating neutralizing antibody levels <20-80. All of these mice were protected from challenge with RSSEV or CEEV. Mice that received 10 microl of sera from monkeys vaccinated with the individual DNA vaccines, both DNA vaccines, or a commercial vaccine were partially to completely protected from RSSEV or CEEV challenge. These data suggest that DNA vaccines may offer protective immunity to primates similar to that obtained with a commercial inactivated-virus vaccine.     

In vitro immunomodulatory properties of tucaresol in HIV infection

The immunomodulatory properties of tucaresol (compound 589C80) were tested on in vitro antigen- and mitogen-stimulated proliferation and cytokine production by peripheral blood mononuclear cells (PBMC) of HIV-infected individuals and healthy controls (HC). Results showed that tucaresol: (1) increases influenza A virus-, gp 160 peptide-, and HLA alloantigen-stimulated proliferation as well as interleukin (IL)-2 and interferon gamma (IFN gamma) production by PBMC of HIV-infected individuals with higher CD4 counts (>500/microl) but had only a marginal immunomodulatory effect on PBMC of patients with lower CD4 counts (<500/microl); (2) did not modify IL-10 production; (3) augmented CD25 expression on mitogen-stimulated T cells of HC but not of HIV-infected individuals; and (4) marginally increased CTL activity. The immunomodulatory properties of tucaresol were confirmed by PCR analyses; additional data showed that tucaresol costimulated CD3-dependent triggering of T cells and that this stimulation was independent of CD28 costimulation. The immunomodulatory effects of tucaresol on T cell functions are characterized by a bell-shaped dose response curve; the action of the compound is optimal in the 100 to 300 microM range. Analyses of mitogen-stimulated apoptosis demonstrated that the lack of effect of tucaresol at higher doses is not the result of increased cell death, suggesting a role of functional impairment. These data confirm that tucaresol can stimulate T helper cell function and enhance the production of type 1 cytokines, thus eliciting cell-mediated immunity, and warrant its potential utility in the therapy of HIV infection.     

Infection of cynomolgus macaques with a recombinant monkeypox virus encoding green fluorescent protein

Monkeypox virus (MPXV) causes a vesiculopustular rash illness resembling smallpox in humans and produces a similar disease in nonhuman primates. To enhance the ability of researchers to study experimental MPXV infections, we inserted a gene encoding green fluorescent protein (GFP) into Monkeypox virus Zaire-79. Wild-type and MPXV-GFP replicated with similar kinetics in cell culture and caused a similar disease when injected intravenously into cynomolgus macaques. In MPXV-GFP-infected animals, examination under fluorescent light facilitated the identification of skin lesions during disease development and internal sites of replication at necropsy. MPXV-GFP could improve the quantitative assessment of antiviral therapy and vaccine efficacy.     

British Association of Critical Care Nurses position statement on prescribing in critical care

Background: Nurses in the UK are now one group of non-medical staff who can prescribe. This practice is evolving for critical care nursing staff who care for critically ill patients during their stay in hospital through ward and outpatient follow-up after admission to critical care.
Aim: The purposes of this paper were to present existing information regarding prescribing to support nurses in critical care currently prescribing and to inform those who are intending to prescribe.
Methods: To develop the position statement, a search of the literature was conducted using key databases. To ascertain the current level and type of prescribing in critical care, a short questionnaire was sent by email to British Association of Critical Care Nursing members, and the results of this are presented in Appendix A.
Outcomes/Results: Evidence was found in relation to the history, context in critical care, educational requirements and issues of consent related to non-medical prescribing.
Conclusions: The position statement is based upon evidence from the literature, National Health Service policy and the Nursing and Midwifery Council regulations. It takes account of the critical care patient pathway before, during and after an admission to critical care.     

Sequence variation within the neuropeptide Y gene and obesity in Mexican Americans

OBJECTIVE: Recently, we reported evidence for linkage between neuropeptide Y (NPY) and both obesity and several obesity-related quantitative measures in a sample of Mexican Americans from Starr County, Texas. The purpose of this study was to investigate putative variation within the coding and promoter regions of NPY. RESEARCH METHODS AND PROCEDURES: Five young, obese individuals (body mass index [BMI] 33 to 45 kg/m2, age 14 to 30 years); five adult, lean individuals (BMI 20 to 26 kg/m2, age 39 to 65 years); and five sibling pairs sharing no alleles that were identical by descent at a marker locus proximal to NPY were selected for fluorescence-based sequencing of approximately 1100 base pairs (bp) immediately 5' from the start site and all four exons of NPY. We identified a total of eight variant sites, including a 2-bp insertion/deletion (I/D) within a putative negative regulatory region (-880I/D) and a 17-bp deletion at the exon 1/intron 1 junction (69I/D). The -880I/D and 69I/D variants were typed in a separate random sample of Mexican Americans (N = 914) from Starr County, Texas. RESULTS: Analyses of variance resulted in a significant association between -880I/D and waist-to-hip ratio (p = 0.041) in the entire sample and between -880I/D and BMI (p = 0.031), abdominal circumference (p = 0.044), and waist-to-hip ratio (p = 0.041) in a non-obese subsample (BMI < 30 kg/m2, n = 594). The 69I/D variant was observed in only one pedigree and does not appear to segregate with obesity within this pedigree. DISCUSSION: This study reports newly identified common human sequence variation within the regulatory and coding sequence of NPY. Several variants were observed, and of those tested, the -880I/D promoter region variant may influence body fat patterning in non-obese individuals but does not appear to play a major role in the etiology of common forms of obesity in this population.     

Performance and sensitivity of modern home pregnancy tests

We have studied the performance of three currently available home pregnancy tests. The Advance is a test based on monoclonal antibodies in an enzyme immunoassay format that specifically detects hCG in urine. The positive results are determined by the presence of a blue color in a color stick (30 minutes). The Daisy 2 and FACT are tests based on monoclonal antibodies that specifically detect hCG in urine in a hemagglutination inhibition assay; positive results are demonstrated by the deposition of a dot ring at the bottom of the test tube (45 minutes). Thirty-five patients who eventually became pregnant in that cycle collected the first morning urine specimens from day of ovulation (determined by ultrasound) in artificial insemination donor (AID) cycles, or from the date of gamete intrafallopian transfer (GIFT) for 16 consecutive days. In each specimen, the Advance, Daisy 2, and FACT tests were performed; in addition, beta-hCG levels were determined by radioimmunoassay. All the home pregnancy tests studied can detect pregnancy as early as 9 or 10 days post-conception, and they give positive results at the time of the expected onset of menses in 70%, 95%, and 88% of cases for Daisy 2, Advance, and FACT, respectively. The sensitivity of the these home pregnancy tests was determined to be about 200 mIU/mL of urine after correlating their positive or negative results with the concentrations of urinary beta-hCG determined by radioimmunoassay.     

Orogenital sex as a cause of nonfatal air embolism in pregnancy

A case of nonfatal air embolism from orogenital sex in the 30th week of pregnancy is described. Because of a delay in the diagnosis, the patient did not receive hyperbaric oxygen therapy until 39 hours after the incident. Severe neurologic dysfunction persisted despite hyperbaric therapy. The world literature on orogenital sex in pregnancy with resultant air embolism is reviewed. The pathophysiology of air embolism and the rationale for hyperbaric oxygen treatment are presented. It is strongly recommended that survivors of this form of air embolism be transferred to hyperbaric facilities as soon as possible to reduce the long-term neurologic sequelae.     

A 9-mo randomized clinical trial comparing fat-substituted and fat-reduced diets in healthy obese men: the Ole Study

BACKGROUND: Dietary fat has been implicated as a risk factor for cardiovascular disease and obesity. OBJECTIVE: We evaluated the effect on body weight, body fat, lipids, glucose, and insulin of replacing dietary fat with olestra in moderately obese men. DESIGN: Forty-five healthy overweight men were randomly assigned to 1 of 3 diets: control diet (33% fat), fat-reduced diet (25% fat), or fat-substituted diet (one-third of dietary fat replaced by olestra to achieve a diet containing 25% metabolizable fat). Body fat was measured by dual-energy X-ray absorptiometry and visceral and subcutaneous abdominal fat by computed tomography. RESULTS: Thirty-six men completed the 9-mo study. Body weight and body fat in the fat-substituted group declined by a mean (+/- SEM) of 6.27 +/- 1.66 and 5.85 +/- 1.34 kg, respectively, over 9 mo compared with 3.8 +/- 1.34 and 3.45 +/- 1.0 kg in the control group and 1.79 +/- 0.81 and 1.68 +/- 0.75 kg in the fat-reduced diet group. At 9 mo, the mean difference in body fat between the fat-reduced and fat-substituted groups was -4.19 +/- 1.19 kg (95% CI: -6.57, -1.81), that between the control and fat-substituted groups was -2.55 +/- 1.21 kg (-0.13, -4.97), and that between the control and fat-reduced groups was 1.63 +/- 1.18 kg (3.96, -0.70). The men eating the fat-reduced diet asked for almost no extra foods, in contrast with the significantly higher requests (P < 0.05) from both of the other 2 groups. CONCLUSION: Replacement of dietary fat with olestra reduces body weight and total body fat when compared with a 25%-fat diet or a control diet containing 33% fat.     

Prediction of body fat in 12-y-old African American and white children: evaluation of methods

BACKGROUND: The prevalence of obesity is increasing in children. Validation of methods of predicting fatness in African American and white children could help to identify children at high risk. OBJECTIVE: We assessed published methods for determining body fat in 12-y-old male and female white and African American schoolchildren. DESIGN: The body fat of 114 children was measured with the use of dual-energy X-ray absorptiometry, underwater weighing (densitometry), measurement of skinfold thicknesses, isotope dilution (H(2)(18)O), and bioelectrical impedance analysis. Formulas derived from these data and from published reports were compared by using the Bland-Altman approach. RESULTS: Calculation of percentage of body fat by using an equation predicting body fat in kg and dividing by the current weight was the criterion method against which the other methods were compared. Four-compartment models had the smallest variability across the range of body fat, and 2 of these models differed from the criterion method by 1-2%. Six methods (the Pennington 4-compartment model, the Wells et al 4-compartment model, the isotope dilution model, dual-energy X-ray absorptiometry, the Pennington skinfold thickness model, and the Pennington density model) provided specificity > 90%, an estimate of body fat that was within the 95% CI of the criterion method, and a difference from the criterion method that was < +/- 2%. Bioelectrical impedance analysis was the least acceptable method. CONCLUSIONS: A 4-compartment model in which body fat in kg is divided by current body weight and multiplied by 100 provides the best estimate of percentage of body fat. The isotope dilution and body density models provide estimates within 2% of the estimate provided by the 4-compartment model. Other models do less well.