Structure of poliovirus replicative intermediate RNA. Electron microscope analysis of RNA cross-linked in vivo with psoralen derivative.

The structure of the poliovirus replicative intermediate RNA was examined by electron microscopy after cross-linking in vivo with 4'-aminomethyl-4,5',8-trimethylpsoralen. After purification from infected cells, undenatured RI appeared as a double-stranded backbone of genome length, with an average of three (and occasionally up to eight) nascent, single-stranded tails. After denaturation, however, only single strands of heterogeneous length were visualized, indicating that the RI in the cell contains little or no duplex structure, and thus nascent chains are only transiently hydrogen-bonded to their template over short regions. The double-stranded backbone of undenatured RI, observed previously by others and in these experiments, is due to collapse of complementary chains during the deproteinization and purification procedures. The effectiveness of the in vivo cross-linking procedure was demonstrated by the complete inhibition of viral RNA synthesis in treated cells and by direct binding of [3H]AMT to RI molecules in vivo. Mature polio virions are impermeable to AMT; however, growth of virus in cells incubated with AMT in the dark resulted in normal yields of virus particles containing RNA genomes, whose infectivity could be subsequently photo-inactivated. The frequency of AMT-induced cross-linking was determined by analyses of double-stranded poliovirus RNA (RF). Cross-linking in vitro followed by spreading for electron microscopy under denaturing conditions yielded bubbled duplex structures with a minimum of one interstrand cross-link per 80 base-pairs. RF cross-linked in vivo also showed extensive cross-linking, decreased about fivefold from the in vitro cross-linked value. Thus, the failure to detect cross-linked RI under these conditions indicates that extensive base-pairing does not exist in vivo.

Efficacy of repeat intravenous dosing of ondansetron in controlling postoperative nausea and vomiting: a randomized, double-blind, placebo-controlled multicenter trial.

STUDY OBJECTIVES: To compare repeat intravenous (i.v.) dosing of ondansetron 4 mg with placebo for the treatment of postoperative nausea and vomiting (PONV) in patients for whom prophylactic, preoperative ondansetron 4 mg i.v. was inadequate DESIGN: Randomized, double-blind, placebo-controlled study. SETTING: Ten outpatient surgical centers in the United States. PATIENTS: 2,199 male and female ASA physical status I, II, and III patients > or = 12 years old scheduled to undergo outpatient surgical procedures and receive nitrous oxide-based general anesthesia. INTERVENTIONS: Ondansetron 4 mg i.v. was administered to all patients before induction of general anesthesia. Patients who experienced PONV or requested antiemetic therapy within 2 hours after discontinuation of inhaled anesthesia were randomized (1:1) to either a repeat i.v. ondansetron 4 mg dose or placebo. MEASUREMENTS AND MAIN RESULTS: Of the 2,199 patients prophylactically treated with ondansetron 4 mg before anesthesia induction, 1,771 (80.5%) did not experience PONV or request antiemetic therapy during the 2 hours following discontinuation of anesthesia. Of the 428 patients who experienced PONV or requested antiemetic therapy during the same period, and were randomized to additional treatment (214 randomized to ondansetron, 214 randomized to placebo), the incidence of complete response (no emesis, no rescue medication, no study withdrawal) was similar for both ondansetron-randomized and placebo-randomized groups for the 2-hour (34% and 43%, respectively, p = 0.074) and 24-hour (28% and 32%, respectively, p = 0.342) postrandomization study periods. Repeat ondansetron dosing was not more effective than placebo in controlling either postoperative emesis or the severity/duration of postoperative nausea. The administration of an additional dose of ondansetron 4 mg postoperatively did not result in an increased incidence of adverse effects. CONCLUSIONS: In patients for whom preoperative prophylaxis with ondansetron 4 mg i.v. is not successful, a repeat dose of ondansetron 4 mg i.v. in the postanesthesia care unit does not appear to offer additional control of PONV.

Restriction of translation of capped mRNA in vitro as a model for poliovirus-induced inhibition of host cell protein synthesis: relationship to p220 cleavage.

Poliovirus infection of HeLa cells results in a rapid inhibition of host protein synthesis by a mechanism that does not affect the translation of poliovirus RNA. It has been suggested that this virus-induced translational control results from inactivation of the cap-binding protein complex, and it has been shown that the 220-kilodalton component(s) (p220) of the cap-binding protein complex is cleaved in infected HeLa cells to form antigenically related polypeptides of 100 to 130 kilodaltons. We have previously described an activity in infected cells that specifically restricts translation of capped mRNA in rabbit reticulocyte lysates. Here, we describe further refinements and characterization of restriction assay. We determined that the assay is a good in vitro model for study of host cell shutoff by several criteria: (i) translation was inhibited in both instances at the step involving mRNA binding to ribosomes; (ii) translation of capped mRNA was specifically inhibited, whereas translation of poliovirus RNA was not; (iii) restriction activity appeared in infected cells with kinetics which parallel host cell shutoff; and (iv) restriction activity, like the specific inhibition of host translation, appeared in cells infected in the presence of guanidine-HCl. The restricting activity was partially purified from poliovirus-infected cells and was compared with the virus-induced p220 cleavage activity. Both activities copurified through numerous cell fractionation and biochemical fractionation procedures. However, specific restriction of capped mRNA translation in reticulocyte lysates occurred without complete cleavage of the endogenous p220.

Sugarless gum chewing before surgery does not increase gastric fluid volume or acidity.

Patients occasionally arrive in the operating suite chewing gum despite instructions to avoid oral intake for a specific number of hours before surgery. Some anaesthetists are hesitant to proceed with these patients fearing an increase in gastric volume and acidity. This study was undertaken to determine if gum chewing increased gastric volume and acidity. Seventy seven patients were recruited and informed consent obtained. Thirty-one patients who fasted overnight were randomly assigned either to serve as control (Group 1) or to chew sugarless gum prior to anaesthesia (Group 2). The remaining 46 patients fasted overnight but were given sugarless gum and allowed to chew it until immediately before induction of anaesthesia if they desired (Group 3). Volume and pH of gastric content were determined immediately after induction of anaesthesia and tracheal intubation. Results revealed mean values (range) of gastric volume for Group 1-26 ml (9-60), Group 2-40 ml (5-93), and Group 3-28 ml (4-65). Mean values for pH (range) were Group 1-1.8 (1.0-4.6), Group 2-1.6 (1.3-1.9), Group 3-1.7 (1.0-4.4). There was no difference between groups in terms of gastric volume or pH. In addition, there was no relationship between gastric content and the length of time from gum discard to induction or the length of time gum was chewed. In conclusion, the data suggest that induction of anaesthesia is safe and surgery does not need to be delayed if a patient arrives in the OR chewing sugarless gum.

Effect of obesity on safe duration of apnea in anesthetized humans.

Obese patients have a decreased functional residual capacity and, hence, a reduced oxygen supply during periods of apnea. To determine whether obese patients are at greater risk of developing hypoxemia during induction of anesthesia than patients of normal weight, 24 patients undergoing elective surgical procedures were studied. Group 1 (normal) were within 20% of their ideal body weight. Group 2 (obese) were more than 20% but less than 45.5 kg over ideal body weight. Group 3 (morbidly obese) were more than 45.5 kg over ideal body weight. Patients were preoxygenated for 5 min or until expired nitrogen was less than 5%. After induction of anesthesia and muscle relaxation the patients were allowed to remain apneic until arterial saturation as measured by pulse oximetry reached 90%. The time taken for oxygen saturation to decrease to 90% was 364 +/- 24 s in group 1, 247 +/- 21 s in group 2, and 163 +/- 15 s in group 3; these times are significantly different at P less than 0.05 between groups. Regression analysis of the data demonstrated a significant negative linear correlation (r = -0.83) between time to desaturation and increasing obesity. These results show that obese patients are at an increased risk of developing hypoxemia when apneic.