Analysis of the transcriptome of the protozoan Theileria parva using MPSS reveals that the majority of genes are transcriptionally active in the schizont stage.

Massively parallel signature sequencing (MPSS) was used to analyze the transcriptome of the intracellular protozoan Theileria parva. In total 1,095,000, 20 bp sequences representing 4371 different signatures were generated from T.parva schizonts. Reproducible signatures were identified within 73% of potentially detectable predicted genes and 83% had signatures in at least one MPSS cycle. A predicted leader peptide was detected on 405 expressed genes. The quantitative range of signatures was 4-52,256 transcripts per million (t.p.m.). Rare transcripts (<50 t.p.m.) were detected from 36% of genes. Sequence signatures approximated a lognormal distribution, as in microarray. Transcripts were widely distributed throughout the genome, although only 47% of 138 telomere-associated open reading frames exhibited signatures. Antisense signatures comprised 13.8% of the total, comparable with Plasmodium. Eighty five predicted genes with antisense signatures lacked a sense signature. Antisense transcripts were independently amplified from schizont cDNA and verified by sequencing. The MPSS transcripts per million for seven genes encoding schizont antigens recognized by bovine CD8 T cells varied 1000-fold. There was concordance between transcription and protein expression for heat shock proteins that were very highly expressed according to MPSS and proteomics. The data suggests a low level of baseline transcription from the majority of protein-coding genes.

Using the transcriptome to annotate the genome revisited: application of massively parallel signature sequencing (MPSS).

Transcriptome analysis can provide useful data for refining genome sequence annotation. Application of massively parallel signature sequencing (MPSS) revealed reproducible transcription, in multiple MPSS cycles, from 73% of computationally predicted genes in the Theileria parva schizont lifecycle stage. Signatures spanning consecutive exons confirmed 142 predicted introns. MPSS identified 83 putative genes, >100 codons overlooked by annotation software, and 139 potentially incorrect gene models (with either truncated ORFs or overlooked exons) by interfacing signature locations with stop codon maps. Twenty representative models were confirmed as likely to be incorrect using reverse transcription PCR amplification from independent schizont cDNA preparations. More than 50% of the 60 putative single copy genes in T. parva that were absent from the genome of the closely related T. annulata had MPSS signatures. This study illustrates the utility of MPSS for improving annotation of small, gene-rich microbial eukaryotic genomes.

Critical care unit organization and patient outcomes.

The delivery of critical care medicine has seen many advances and changes over a relatively short period of time. This article explores some of the models of critical care delivery and the implications of these models on patient outcomes.

Unintentional human exposure to tilmicosin (Micotil 300).

BACKGROUND: Tilmicosin phosphate is a macrolide antibiotic that is used to treat cattle for pathogens that cause Bovine Respiratory Disease. CASE REPORT: A 28-year-old man with no prior history of heart disease developed severe chest pain, inverted T waves, and intraventricular conduction delay on EKG and mild elevation of cardiac enzymes 5 hours after unintentional injection of less than half of a 12cc syringe filled with Micotil 300 (tilmicosin phosphate 300 mg/mL, propylene glycol 25%, phosphoric acid, water for injection). The patient made an uneventful recovery after hospitalization. CONCLUSIONS: This case provides evidence that unintentional injection of tilmicosin can cause cardiac symptoms and laboratory evidence of myocardial injury. Tilmicosin should always be administered by properly trained personnel who are using techniques designed to reduce the risk of accidental self-injection.