Study of the impact of WHSC1 and CEP55 genes silencing in myxofibrosarcoma cells

Myxofibrosarcoma is one of the most common soft tissue sarcomas, contributing to more than 5% of all adult sarcomas. This neoplasm most often develops in the dermis or subcutaneous tissue of the extremities. However, it may also be deep-seated and arise in other locations such as the head, neck or trunk (1,2). Although adult sarcomas have received more attention in recent years, myxofibrosarcoma remains vastly understudied (about 400 articles referenced on PubMed since the 1950s). Therefore, further knowledge about the tumour-initiating capacity of primary myxofibrosarcoma tumour cells is greatly needed to identify targets that could be susceptible for specific treatments. By using gene expression microarrays, two genes (WHSC1 and CEP55) were found to be overexpressed in the more aggressive and metastatic primary myxofibrosarcoma cell population “SpA”, as compared to the non-metastatic cell population “DMEM” derived from the same primary myxofibrosarcoma tumour. The present study was aimed at assessing the importance of these two genes for the survival of primary myxofibrosarcoma cells. These genes were silenced using the short hairpin RNA (shRNA) technique in two cell populations, cultured in different conditions (DMEM and SpA). Cell cultures with stable WHSC1 or CEP55 depletion were then injected into the renal capsule of mice to evaluate their tumorigenic capability. Although tumour formation in vivo was decreased to some extent in shRNA-treated cells, as compared to control cells, the tumour-forming ability was not abolished. As it could be partly due to residual protein expression, we designed a complete gene knockout by utilizing another silencing technique, the genome editing CRISPR CAS9 system. To date, these constructs targeting WHSC1 and CEP55, respectively, have not yet been evaluated in the target cells. This study may open the way to a better understanding of the tumorigenesis of myxofibrosarcoma, as well as the role of WHSC1 and CEP55 genes, by studying the impact of their silencing both in vitro and in vivo

Evidence that conflict regarding size of haemodynamic response to interventricular delay optimization of cardiac resynchronization therapy may arise from differences in how atrioventricular delay is kept constant.

Aims: Whether adjusting interventricular (VV) delay changes haemodynamic efficacy of cardiac resynchronization therapy (CRT) is controversial, with conflicting results. This study addresses whether the convention for keeping atrioventricular (AV) delay constant during VV optimization might explain these conflicts. / Method and results: Twenty-two patients in sinus rhythm with existing CRT underwent VV optimization using non-invasive systolic blood pressure. Interventricular optimization was performed with four methods for keeping the AV delay constant: (i) atrium and left ventricle delay kept constant, (ii) atrium and right ventricle delay kept constant, (iii) time to the first-activated ventricle kept constant, and (iv) time to the second-activated ventricle kept constant. In 11 patients this was performed with AV delay of 120 ms, and in 11 at AV optimum. At AV 120 ms, time to the first ventricular lead (left or right) was the overwhelming determinant of haemodynamics (13.75 mmHg at ±80 ms, P < 0.001) with no significant effect of time to second lead (0.47 mmHg, P = 0.50), P < 0.001 for difference. At AV optimum, time to first ventricular lead again had a larger effect (5.03 mmHg, P < 0.001) than time to second (2.92 mmHg, P = 0.001), P = 0.02 for difference. / Conclusion: Time to first ventricular activation is the overwhelming determinant of circulatory function, regardless of whether this is the left or right ventricular lead. If this is kept constant, the effect of changing time to the second ventricle is small or nil, and is not beneficial. In practice, it may be advisable to leave VV delay at zero. Specifying how AV delay is kept fixed might make future VV delay research more enlightening

A wideband radar for high-resolution mapping of near-surface internal layers in glacial ice

©2004 IEEE. Personal use of this material is permitted. However, permission to reprint/republish this material for advertising or promotional purposes or for creating new collective works for resale or redistribution to servers or lists, or to reuse any copyrighted component of this work in other works must be obtained from the IEEE.Snow accumulation rate is an important parameter in determining the mass balance of polar ice sheets. Accumulation rate is currently determined by analyzing ice cores and snow pits. Inadequate sampling of the spatial variations in the ice sheet accumulation has resulted in accumulation rate uncertainties as large as 24%. We designed and developed a 600-900-MHz airborne radar system for high-resolution mapping of the near-surface internal layers for estimating the accumulation rate of polar ice sheets. Our radar system can provide improved spatial and temporal coverage by mapping a continuous profile of the isochronous layers in the ice sheet. During the 2002 field season in Greenland, we successfully mapped the near-surface layers to a depth of 200 m in the dry-snow zone, 120 m in the percolation zone, and 20 m in the melt zone. We determined the water equivalent accumulation rate at the NASA-U_1 site to be 34.9 +/- 5.1 cm/year from 1964 to 1992. This is in close agreement with the ice-core derived accumulation rate of 34.6 cm/year for the same period

Coherent radar ice thickness measurements over the Greenland ice sheet

This is the published version, also available here: http://dx.doi.org/10.1029/2001JD900183.We developed two 150-MHz coherent radar depth sounders for ice thickness measurements over the Greenland ice sheet. We developed one of these using connectorized components and the other using radio frequency integrated circuits (RFICs). Both systems are designed to use pulse compression techniques and coherent integration to obtain the high sensitivity required to measure the thickness of more than 4 km of cold ice. We used these systems to collect radar data over the interior and margins of the ice sheet and several outlet glaciers. We operated both radar systems on the NASA P-3B aircraft equipped with GPS receivers. Radar data are tagged with GPS-derived location information and are collected in conjunction with laser altimeter measurements. We have reduced all data collected since 1993 and derived ice thickness along all flight lines flown in support of Program for Regional Climate Assessment (PARCA) investigations and the North Greenland Ice Core Project. Radar echograms and derived ice thickness data are placed on a server at the University of Kansas (http://tornado.rsl.ukans.edu/Greenlanddata.htm) for easy access by the scientific community. We obtained good ice thickness information with an accuracy of ±10 m over 90% of the flight lines flown as a part of the PARCA initiative. In this paper we provide a brief description of the system along with samples of data over the interior, along the 2000-m contour line in the south and from a few selected outlet glaciers

Remodelling of gap junctions and connexin expression in diseased myocardium

Gap junctions form the cell-to-cell pathways for propagation of the precisely orchestrated patterns of current flow that govern the regular rhythm of the healthy heart. As in most tissues and organs, multiple connexin types are expressed in the heart: connexin43 (Cx43), Cx40 and Cx45 are found in distinctive combinations and relative quantities in different, functionally-specialized subsets of cardiac myocyte. Mutations in genes that encode connexins have only rarely been identified as being a cause of human cardiac disease, but remodelling of connexin expression and gap junction organization are well documented in acquired adult heart disease, notably ischaemic heart disease and heart failure. Remodelling may take the form of alterations in (i) the distribution of gap junctions and (ii) the amount and type of connexins expressed. Heterogeneous reduction in Cx43 expression and disordering in gap junction distribution feature in human ventricular disease and correlate with electrophysiologically identified arrhythmic changes and contractile dysfunction in animal models. Disease-related alterations in Cx45 and Cx40 expression have also been reported, and some of the functional implications of these are beginning to emerge. Apart from ventricular disease, various features of gap junction organization and connexin expression have been implicated in the initiation and persistence of the most common form of atrial arrhythmia, atrial fibrillation, though the disparate findings in this area remain to be clarified. Other major tasks ahead focus on the Purkinje/working ventricular myocyte interface and its role in normal and abnormal impulse propagation, connexin-interacting proteins and their regulatory functions, and on defining the precise functional properties conferred by the distinctive connexin co-expression patterns of different myocyte types in health and disease

A systematic approach to designing reliable VV optimization methodology: Assessment of internal validity of echocardiographic, electrocardiographic and haemodynamic optimization of cardiac resynchronization therapy

AbstractBackgroundIn atrial fibrillation (AF), VV optimization of biventricular pacemakers can be examined in isolation. We used this approach to evaluate internal validity of three VV optimization methods by three criteria.Methods and resultsTwenty patients (16 men, age 75±7) in AF were optimized, at two paced heart rates, by LVOT VTI (flow), non-invasive arterial pressure, and ECG (minimizing QRS duration). Each optimization method was evaluated for: singularity (unique peak of function), reproducibility of optimum, and biological plausibility of the distribution of optima.The reproducibility (standard deviation of the difference, SDD) of the optimal VV delay was 10ms for pressure, versus 8ms (p=ns) for QRS and 34ms (p<0.01) for flow.Singularity of optimum was 85% for pressure, 63% for ECG and 45% for flow (Chi2=10.9, p<0.005).The distribution of pressure optima was biologically plausible, with 80% LV pre-excited (p=0.007). The distributions of ECG (55% LV pre-excitation) and flow (45% LV pre-excitation) optima were no different to random (p=ns).The pressure-derived optimal VV delay is unaffected by the paced rate: SDD between slow and fast heart rate is 9ms, no different from the reproducibility SDD at both heart rates.ConclusionsUsing non-invasive arterial pressure, VV delay optimization by parabolic fitting is achievable with good precision, satisfying all 3 criteria of internal validity. VV optimum is unaffected by heart rate. Neither QRS minimization nor LVOT VTI satisfy all validity criteria, and therefore seem weaker candidate modalities for VV optimization. AF, unlinking interventricular from atrioventricular delay, uniquely exposes resynchronization concepts to experimental scrutiny

Cardiac resynchronization therapy: mechanisms of action and scope for further improvement in cardiac function.

Aims: Cardiac resynchronization therapy (CRT) may exert its beneficial haemodynamic effect by improving ventricular synchrony and improving atrioventricular (AV) timing. The aim of this study was to establish the relative importance of the mechanisms through which CRT improves cardiac function and explore the potential for additional improvements with improved ventricular resynchronization. Methods and Results: We performed simulations using the CircAdapt haemodynamic model and performed haemodynamic measurements while adjusting AV delay, at low and high heart rates, in 87 patients with CRT devices. We assessed QRS duration, presence of fusion, and haemodynamic response. The simulations suggest that intrinsic PR interval and the magnitude of reduction in ventricular activation determine the relative importance of the mechanisms of benefit. For example, if PR interval is 201 ms and LV activation time is reduced by 25 ms (typical for current CRT methods), then AV delay optimization is responsible for 69% of overall improvement. Reducing LV activation time by an additional 25 ms produced an additional 2.6 mmHg increase in blood pressure (30% of effect size observed with current CRT). In the clinical population, ventricular fusion significantly shortened QRS duration (Δ-27 ± 23 ms, P < 0.001) and improved systolic blood pressure (mean 2.5 mmHg increase). Ventricular fusion was present in 69% of patients, yet in 40% of patients with fusion, shortening AV delay (to a delay where fusion was not present) produced the optimal haemodynamic response. Conclusions: Improving LV preloading by shortening AV delay is an important mechanism through which cardiac function is improved with CRT. There is substantial scope for further improvement if methods for delivering more efficient ventricular resynchronization can be developed. Clinical Trial Registration: Our clinical data were obtained from a subpopulation of the British Randomised Controlled Trial of AV and VV Optimisation (BRAVO), which is a registered clinical trial with unique identifier: NCT01258829, https://clinicaltrials.gov