Candida albicans strain maintenance, replacement, and microvariation demonstrated by multilocus sequence typing.

We typed 165 Candida albicans isolates from 44 different sources by multilocus sequence typing (MLST) and ABC typing of rRNA genes and determined their homozygosity or heterozygosity at the mating-type-like locus (MTL). The isolates represented pairs or larger sets from individual sources, which allowed the determination of strain diversity within patients. A comparison of replicate sequence data determined a reproducibility threshold for regarding isolates as MLST indistinguishable. For 36 isolate sets, MLST and ABC typing showed indistinguishable or highly related strain types among isolates from different sites or from the same site at different times from each patient. This observation included 11 sets with at least one isolate from a blood culture and a nonsterile site from the same patient. For one patient, strain replacement was evidenced in the form of two sets of isolates from different hospital admissions where the strain types within each set were nearly identical but where the two sets differed both by MLST and ABC typing. MLST therefore confirms the existing view of C. albicans strain carriage. Microvariation, evidenced as small differences between MLST types, resulted in most instances from a loss of heterozygosity at one or more of the sequenced loci. Among isolate sets that showed major strain type differences, some isolates could be excluded as likely examples of handling errors during storage. However, for a minority of isolates, intermittent differences in ABC type for tightly clustered MLST types and intermittent appearances of MTL homozygosity lead us to propose that some C. albicans isolates, or all isolates under yet-to-be-determined conditions, maintain a high level of genetic diversity by mechanisms such as recombination, gene conversion, or chromosomal ploidy change.

Cloning and expression of single chain antibody fragments in Escherichia coli and Pichia pastoris.

The potential of recombinant antibody fragments is likely to be fulfilled only if they can be produced routinely at high concentrations. We have compared the ability of Escherichia coli and Pichia pastoris to produce functional recombinant single chain antibody (scAb) fragments. Two scAb fragments were expressed, an antihuman type V acid phosphatase (TRAP) and an anti-Pseudomonas aeruginosa lipoprotein I. We report here that, while expression from P. pastoris resulted in a significantly increased level of expression of the anti-TRAP scAb compared to E. coli, neither fragment was able to bind its target antigen as well as the bacterial product.

A comparison of eltanolone and propofol anaesthesia for minor gynaecological surgery.

We have used eltanolone and propofol to induce anaesthesia and by intermittent injection to supplement nitrous oxide during maintenance in 67 patients undergoing minor gynaecological surgery. This study was a controlled randomized phase III with two parallel groups and a blinded assessment of post-operative recovery. Complications during induction and maintenance were few. Urticaria occurred in two patients given eltanolone and in none of those given propofol. Time to orientation and time to eye opening were significantly shorter in the propofol group (P < 0.001). Systolic and diastolic blood pressure decreased in both groups but the decrease was significantly greater in the propofol group (P < 0.005 systolic blood pressure; P < 0.001 diastolic blood pressure). Heart rate decreased significantly in the propofol group (P < 0.002). We conclude that eltanolone anaesthesia is associated with a greater haemodynamic stability than propofol anaesthesia but that early recovery occurs more slowly.

Comparison of induction and recovery between sevoflurane and halothane supplementation of anaesthesia in children undergoing outpatient dental extractions.

We have compared sevoflurane and halothane in a double-blind controlled study for supplementation of nitrous oxide and oxygen anaesthesia in 80 children undergoing dental extraction as outpatients. Induction of anaesthesia was more rapid in those who received sevoflurane compared with those who received halothane (89 s compared with 127 s for loss of eyelash reflex). In both groups, mean duration of administration of anaesthesia was less than 4 min. Those who received sevoflurane were slower to awaken (167 s compared with 102 s), although discharge times from hospital were similar. The incidence of complications during induction and maintenance was low in both groups and return to normal appetite and activity occurred in the majority of children on the same day. More children who received halothane suffered nausea after leaving hospital. We conclude that sevoflurane is a suitable alternative to halothane, with more rapid induction of anaesthesia, but in these short procedures, awakening time was slower than after halothane.