Rat brain creatine kinase (CKB) gene expression is highest in the brain but is also detectable at lower levels in some other tissues. In the brain, the CKB enzyme is thought to be involved in the regeneration of ATP necessary for transport of ions and neurotransmitters. To understand the molecular events that lead to high CKB expression in the brain, we have determined the steady-state levels of CKB mRNA in homogeneous cultures of primary rat brain astrocytes, oligodendrocytes, and neurons. Northern blot analysis showed that whereas the 1.4-kb CKB mRNA was detectable in neurons, the level was about 17-fold higher in oligodendrocytes and 15-fold higher in astrocytes. The blots were hybridized with a CKB-specific 32P-antisense RNA probe, complementary to the 3' untranslated sequence of CKB, which hybridizes to CKB mRNA but not CKM mRNA. Also, the 5' and 3' ends of CKB mRNA from the glial cells were mapped, using exon-specific antisense probes in the RNase-protection assay, and were found to be the same in astrocytes and oligodendrocytes. This indicated that (a) the site of in vivo transcription initiation in astrocytes and oligodendrocytes was directed exclusively by the downstream, nonconcensus TTAA sequence at -25 bp in the CKB promoter that is also utilized by all other cell types that express CKB and (b) the 3' end of mature CKB mRNA was the same in astrocytes and oligodendrocytes. In addition, there was no detectable alternate splicing in exon 1, 2, or 8 of CKB mRNA in rat astrocytes and oligodendrocytes.(ABSTRACT TRUNCATED AT 250 WORDS)

No abstract

We have recently shown that while brain creatine kinase (CKB) mRNA was detectable in RNA from cultured primary rat brain neurons, CKB mRNA was about 15-fold higher in primary astrocytes and 17-fold higher in oligodendrocytes (Molloy et al., J Neurochem 59:1925-1932, 1992). To begin to understand the molecular mechanisms responsible for brain glial cells containing the highest levels of CKB mRNA in the body, we have examined the expression of rat CKB mRNA in established C6 glioma cells. RNase-protection analysis showed the endogenous CKB mRNA levels in exponentially growing C6 were high and measured 50% of that in total RNA from rat brain lysate and 60% of that in cultured primary astrocytes and oligodendrocytes. The 5' and 3' ends of CKB mRNA in C6 were mapped to the same nucleotides as CKB mRNA from rat brain, indicating that the sites of in vivo transcription initiation and termination/polyadenylation of CKB mRNA in C6 are the same as in total rat brain RNA. The level of CKB enzyme activity in C6 whole cell lysates was among the highest of the glial cell lines which we measured. All creatine kinase enzyme activity present in C6 was found in the dimeric CKB isoform (BB), which is characteristic of CKB expression in the brain. A 2.9 kb gene fragment containing the basal CKB promoter and far-upstream 5' sequences was cloned upstream of the chloramphenicol acetyltransferase (CAT) gene and transfected into C6 cells. CAT activity was readily detectable in C6 and mapping of the 5' end of the CAT mRNA showed that transcription was directed from the correct initiation site. Since we found C6 cells were difficult to transfect, conditions were established which both maximized transfection efficiency and maintained normal C6 cell morphology. These results should permit the future identification of the nuclear trans-acting factors and the cognate cis-acting regulatory elements responsible for high CKB mRNA expression in brain glial cells.

Background Preoperative templating software and intraoperative navigation have the potential to impact baseplate augmentation utilization and increase screw length for baseplate fixation in reverse total shoulder arthroplasty (rTSA). We aimed to assess their impact on the (1) baseplate screw length, (2) number of screws used, and (3) frequency of augmented baseplate use in navigated rTSA. Methods We compared 51 patients who underwent navigated rTSA with 63 controls who underwent conventional rTSA at a single institution. Primary outcomes included the screw length, composite screw length, number of screws used, percentage of patients in whom 2 screws in total were used, and use of augmented baseplates. Results Navigation resulted in the use of significantly longer individual screws (36.7 mm vs. 30 mm, P < .0001), greater composite screw length (84 mm vs. 76 mm, P = .048), and fewer screws (2.5 ± 0.7 vs. 2.8 ± 1, P = .047), as well as an increased frequency of using 2 screws in total (35 of 51 patients [68.6%] vs. 32 of 63 controls [50.8%], P = .047). Preoperative templating resulted in more frequent augmented baseplate utilization (76.5% vs. 19.1%, P < .0001). Conclusion The difference in the screw length, number of screws used, and augmented baseplate use demonstrates the evolving role that computer navigation and preoperative templating play in surgical planning and the intraoperative technique for rTSA.

Shared decision-making (SDM) is the process by which the physician and the patient collaborate to arrive at the evidence-based treatment that best aligns with the patient's individual goals of care. The implementation of SDM practices is especially important when an invasive surgical procedure is among the treatment options. Ideally, the variation in the patient treatment choice would mirror the variability in patient goals rather than variation in a physician's ability to communicate effectively. Potentially aiding in these communication efforts, decision aids are tools gaining popularity for their ability to help physicians facilitate the SDM process with patients. This article is intended to give the practicing orthopaedic surgeon an understanding of the concept of SDM and how it can improve physician-patient communication. We will explore the key elements of SDM, attendant benefits, indications, and barriers to implementation and propose steps to begin the incorporation of SDM into practice.