Isolation of single-copy human genes from a library of yeast artificial chromosome clones

A recently developed cloning system based on the propagation of large DNA molecules as linear, artificial chromosomes in the yeast Saccharomyces cerevisiae provides a potential method of cloning the entire human genome in segments of several hundred kilobase pairs. Most application of this system will require the ability to recover specific sequences from libraries of yeast artificial chromosome clones and to propagate these sequences in yeast without alterations. Two single-copy genes have now been cloned from a library of yeast artificial chromosome clones that was prepared from total human DNA. Multiple, independent isolates were obtained of the genes encoding factor IX and plasminogen activator inhibitor type 2. The clones, which ranged in size from 60 to 650 kilobases, were stable on prolonged propagation in yeast and appear to contain faithful replicas of human DNA.     

Direct measurement of the current-phase relation of a superfluid 3He-B weak link

Direct measurements of the current-phase relation, I versus Deltaphi, for a weak link coupling two reservoirs of B-phase superfluid helium-3 (3He-B) were made over a wide range of temperatures. The weak link consists of a square array of 100-nanometer-diameter apertures. For temperatures T such that T/Tc >/= 0.6 (where Tc is the superfluid transition temperature), I approximately sin(Deltaphi). At lower temperatures, I(Deltaphi) approaches a straight line. Several remarkable phenomena heretofore inaccessible to superconducting Josephson junctions, including direct observation of quantum oscillations and continuous knowledge of Deltaphi, were also observed.     

Controlling microbial contamination on beef and lamb meat during processing

SUMMARY The microbiological quality of carcases, meat and environmental surfaces was evaluated in commercial boning rooms processing beef and lamb. There was considerable variation in the level of microbial contamination on both carcases and meat, with counts ranging from less than 20 to 10⁸/cm² on carcases and to 2 times 10⁷/cm² on meat. The level of microbial contamination on meat was influenced by the level of carcase contamination at boning and by the boning process itself. Carcase contamination was the major determinant of microbiological quality, as more than 70% of carcases had microbial counts greater than 10³/cm². Cutting boards were a major source for microbial dissemination during boning, particularly when carcase counts were less than 10³/cm². If carcases were heavily contaminated, the contamination of processing surfaces was irrelevant in determining microbial loads on meat. Where carcase contamination was at low to moderate levels, the contribution of the boning process to the contamination on meat assumed increased significance. Under these conditions, improved sanitation of cutting surfaces in the boning room resulted in a significant reduction in microbial contamination on the surface of meat. These results can form the basis for ensuring that improvements made in carcase management before boning, to improve microbiological quality, will be preserved through attention to cutting board hygiene during boning.     

Effectiveness of passive gas-scavenging canisters attached to isoflurane anesthesia systems under standard-use conditions in a laboratory animal facility

Chronic exposure to trace levels of waste anesthetic gases has been linked to higher incidences of neurologic and reproductive dysfunction, hepatic and renal toxicity, and neoplasia in health care professionals. We have shown that low levels of isoflurane emissions are likely in conventional laboratory animal treatment rooms during the use of standard anesthesia delivery systems equipped with activated charcoal canisters for passive gas scavenging. In the present study, we surveyed the effectiveness of canisters (attached to well‐maintained precision isoflurane vaporizers) in current use throughout our AAALAC‐accredited laboratory animal facility. Canisters (Omnicon f/air) had been weighed prior to use and then attached to dual‐loop systems (face mask and induction box circuits) from 1 week to 6 months of service. Isoflurane emissions were measured using a pre‐calibrated, portable infrared spectrophotometer by attaching each canister to the face‐mask circuit, occluding the face mask and closing the stopcock to the induction circuit, and running the system at uniform isoflurane concentration (2%) and oxygen flow rate (1 L minute^?1). Samples were taken in animal procedure rooms (size range, 45–80 m³) in which the air turnover rate ranged between 20 and 30 nonrecirculating changes per hour. Nine of the 60 canisters (15%) in current use were found to have exceeded the manufacturer's recommended use‐life (defined as a weight increase of 50 g). Of these nine, seven canisters did not scavenge isoflurane at all (indicated by emissions greatly exceeding 100 ppm). Isoflurane was not detected in the operator's breathing zone under normal use conditions (i.e. gas directed to both circuits at once). Of the 51 canisters that had not exceeded their rated use‐life, 12 (23.5%) exhausted isoflurane at >2 ppm. Our data show that (i) the potential for exposure to waste isoflurane emissions will depend on the configuration of the delivery system and that (ii) enhanced attention to canister surveillance may be warranted even in well‐run facilities.