Molecular signatures for CCN1, p21 and p27 in progressive mantle cell lymphoma

Mantle cell lymphoma (MCL) is a comparatively rare non-Hodgkin’s lymphoma characterised by overexpression of cyclin D1.Many patients present with or progress to advanced stage disease within 3 years. MCL is considered an incurable disease withmedian survival between 3 and 4 years. We have investigated the role(s) of CCN1 (CYR61) and cell cycle regulators inprogressive MCL. We have used the human MCL cell lines REC1 G519 > JVM2 cells by RQ-PCR, depicting a decrease in CCN1expression with disease progression. Investigation of CCN1 isoform expression by western blotting showed that whilst expres-sion of full-length CCN1 was barely altered in the cell lines, expression of truncated forms (18–20 and 28–30 kDa) decreasedwith disease progression. We have then demonstrated that cyclin D1 and cyclin dependent kinase inhibitors (p21CIP1and p27KIP1)are also involved in disease progression. Cyclin D1 was highly expressed in REC1 cells (OD: 1.0), reduced to one fifth in G519cells (OD: 0.2) and not detected by western blotting in JVM2 cells. p27KIP1followed a similar profile of expression as cyclin D1.Conversely, p21CIP1was absent in the REC1 cells and showed increasing expression in G519 and JVM2 cells. Subcellularlocalization detected p21CIP1/p27KIP1primarily within the cytoplasm and absent from the nucleus, consistent with altered roles in treatment resistance. Dysregulation of the CCN1 truncated forms are associated with MCL progression. In conjunction withreduced expression of cyclin D1 and increased expression of p21, this molecular signature may depict aggressive disease andtreatment resistance

Beyond equilibrium climate sensitivity

ISSN:1752-0908ISSN:1752-089

Reliability and accuracy of an automated tracking algorithm to measure controlled passive and active muscle fascicle length changes from ultrasound

Manual tracking of muscle fascicle length changes from ultrasound images is a subjective and time-consuming process. The purpose of this study was to assess the repeatability and accuracy of an automated algorithm for tracking fascicle length changes in the medial gastrocnemius (MG) muscle during passive length changes and active contractions (isometric, concentric and eccentric) performed on a dynamometer. The freely available, automated tracking algorithm was based on the Lucas-Kanade optical flow algorithm with an affine optic flow extension, which accounts for image translation, dilation, rotation and shear between consecutive frames of an image sequence. Automated tracking was performed by three experienced assessors, and within- and between-examiner repeatability was computed using the coefficient of multiple determination (CMD). Fascicle tracking data were also compared with manual digitisation of the same image sequences, and the level of agreement between the two methods was calculated using the coefficient of multiple correlation (CMC). The CMDs across all test conditions ranged from 0.50 to 0.93 and were all above 0.98 when recomputed after the systematic error due to the estimate of the initial fascicle length on the first ultrasound frame was removed from the individual fascicle length waveforms. The automated and manual tracking approaches produced similar fascicle length waveforms, with an overall CMC of 0.88, which improved to 0.94 when the initial length offset was removed. Overall results indicate that the automated fascicle tracking algorithm was a repeatable, accurate and time-efficient method for estimating fascicle length changes of the MG muscle in controlled passive and active conditions