Construction of a quinoa (Chenopodium quinoa Willd.) BAC library and its use in identifying genes encoding seed storage proteins.

Quinoa (Chenopodium quinoa Willd.) is adapted to the harsh environments of the Andean Altiplano region. Its seeds have a well-balanced amino acid composition and exceptionally high protein content with respect to human nutrition. Quinoa grain is a staple in the diet of some of the most impoverished people in the world. The plant is an allotetraploid displaying disomic inheritance (2n=4x=36) with a di-haploid genome of 967 Mbp (megabase pair), or 2C=2.01 pg. We constructed two quinoa BAC libraries using BamHI (26,880 clones) and EcoRI (48,000 clones) restriction endonucleases. Cloned inserts in the BamHI library average 113 kb (kilobase) with approximately 2% of the clones lacking inserts, whereas cloned inserts in the EcoRI library average 130 kb and approximately 1% lack inserts. Three plastid genes used as probes of high-density arrayed blots of 73,728 BACs identified approximately 2.8% of the clones as containing plastid DNA inserts. We estimate that the combined quinoa libraries represent at least 9.0 di-haploid nuclear genome equivalents. An average of 12.2 positive clones per probe were identified with 13 quinoa single-copy ESTs as probes of the high-density arrayed blots, suggesting that the estimate of 9.0x coverage of the genome is conservative. Utility of the BAC libraries for gene identification was demonstrated by probing the library with a partial sequence of the 11S globulin seed storage protein gene and identifying multiple positive clones. The presence of the 11S globulin gene in four of the clones was verified by direct comparison with quinoa genomic DNA on a Southern blot. Besides serving as a useful tool for gene identification, the quinoa BAC libraries will be an important resource for physical mapping of the quinoa genome.

Remifentanil and pulmonary extraction during and after cardiac anesthesia.

We measured the apparent blood clearance and pulmonary extraction ratio of remifentanil in 10 adult patients undergoing elective myocardial revascularization for the first time with hypothermic cardiopulmonary bypass (CPB). Patients received continuous infusions of remifentanil 1.0, 1.5 or 2.0 microg x kg(-1) x min(-1). After surgery, remifentanil was infused at 1.0 microg x kg(-1) x min(-1) in all patients. Remifentanil concentrations were measured in pulmonary and radial artery blood by gas chromatography with high resolution mass spectrometry before and after CPB and 165 min (60 SD) after surgery. Cardiac output was measured by thermodilution at the time of blood sampling. The mean pulmonary extraction ratio of remifentanil was 5.7% (13.1% SD), which was not significantly different from zero. However, pulmonary extraction ratio was related inversely to the pulmonary artery hydrogen ion concentration and directly to the percent of nonionized form of the base in the pulmonary artery. Remifentanil concentrations in pulmonary and radial artery blood were related directly to infusion rate, but not to duration of infusion. There was no evidence of accumulation or sequestration. Mean apparent blood remifentanil clearance was 2.03 L/min (0.35 SD) and, in contrast to remifentanil pulmonary extraction ratio, was related directly to cardiac index and oxygen delivery. Increased tissue perfusion increased blood remifentanil clearance. We found predictable blood remifentanil levels with no evidence of accumulation or pulmonary extraction.

Comparison of DonJoy ankle ligament protector and subtalar sling ankle taping in restricting foot and ankle motion before and after exercise.

Clinicians often must select an appropriate prophylactic ankle support system for their patients from a variety of ankle orthoses and ankle taping configurations. The purpose of this study was to compare the effectiveness of the DonJoy Ankle Ligament Protector and a newly developed ankle taping procedure in restricting foot and ankle motion before and after exercise. Subjects were eight males and eight females who reported no history of ankle injury during the 6 months prior to testing, neurological condition, lower extremity arthritis, lower extremity fracture, or cardiac or balance problems. A Biodex dynamometer and computer were used to impose passive moments and to measure eversion and inversion prior to application of the ankle support systems, following application, and following 10 minutes of figure-of-eight running and 20 unilateral toe raises. Both ankles of each subject were assessed for each ankle support system. Subjects also compared the support systems for comfort, stability, and cosmetic acceptability. Both ankle support systems significantly reduced eversion and inversion following application and following exercise compared with preapplication measurements. Eversion measurements increased significantly following exercise for both ankle support systems compared with postapplication measurements. Inversion displacement following application was greater for the Ankle Ligament Protector than the ankle taping system. The two ankle support systems did not differ significantly following exercise for eversion or inversion measurements. The results may assist clinicians in selecting either of these ankle support systems for use in protection against ankle sprain injury.

Prolonged use of isoflurane in a patient with tetanus.

We describe a patient with tetanus, who received isoflurane for sedation to facilitate controlled mechanical ventilation. Isoflurane was administered for 34 days, resulting in a sustained serum inorganic fluoride ion concentration in excess of 50 mumol litre-1 and a peak serum inorganic fluoride ion concentration of 87 mumol litre-1. Although these concentrations are potentially nephrotoxic, no toxicity was evident clinically.

Evaluation of combinations of interferons and cytotoxic drugs in murine tumor models in vivo.

This investigation was conducted to provide preclinical in vivo tumor response data collected under standardized conditions with a range of clinically useful drugs combined with type I (alpha/beta) or type II (gamma) interferon. Murine tumor models used were P388 leukemia, Meth A sarcoma, and B16 melanoma. Eleven cytotoxic drugs were studied. Interferon combinations with cytosine arabinoside provided consistent indications of activity greater than that of the respective single agents. Doxorubicin and cisplatin each prolonged the time to treatment failure, relative to single-agent results, when they were combined with gamma-interferon in the Meth A and B16 models. Interferon combinations with methotrexate, 6-mercaptopurine, 6-thioguanine, ampligen, suramin, 5-fluorouracil, cyclophosphamide, and vinblastine yielded no evidence of any positive therapeutic interactions under the conditions of this study.

Hormonal effects on L-proline transport in coho salmon (Oncorhynchus kisutch) intestine.

The hormonal control of proline transport in pyloric ceca was studied in regard to the effects of cortisol, growth hormone (GH), epinephrine, and 3-isobutyl-1-methylxanthine (IBMX). Cortisol pellets implanted in yearling freshwater (FW) salmon for 2 weeks elevated plasma cortisol levels six times above that of control fish. The maximal influx (Jmax) and the half-saturation constant (Kt) of proline influx were twofold greater in cortisol-treated fish than the values in controls; the apparent passive permeability coefficient (Pa) was significantly reduced in the former group. FW salmon implanted with GH for 2 weeks showed increased body weight gain and a higher Jmax of proline influx compared with that of control fish. GH treatment resulted in a higher Pa of proline influx as well as in a 30% increase in area-specific intestinal dry weight. Thus, GH and cortisol may play a regulatory role in intestinal amino acid absorption during salmon development. The in vitro effects of epinephrine and the phosphodiesterase inhibitor, IBMX, on short-circuit current (Isc) and proline influx in salmon intestine were examined. Epinephrine (10(-6) M) caused a rapid increase in negative Isc (mucosa, ground). Pyloric ceca preincubated with epinephrine for 30 min showed reduced total proline influx compared with influx in paired control tissues. Epinephrine increased and IBMX decreased the Kt of proline influx; IBMX also reduced Jmax. The possible interaction between the effects of epinephrine and IBMX on ion transport and Na+-coupled proline influx are discussed.