c-myc and bcl-2 modulate p53 function by altering p53 subcellular trafficking during the cell cycle.

We have studied the ability of c-myc and bcl-2 oncogenes to modulate p53 function. Our studies show that coincident expression of human Bcl-2 protein with p53 prolongs survival of murine erythroleukemia cells. This effect was associated with a loss of the G1 specificity of p53-mediated cell cycle arrest. Furthermore, we found that the c-myc and bcl-2 genes cooperate to inhibit p53 functions. Coexpression of bcl-2 and c-myc can totally overcome p53-induced apoptosis and cell cycle arrest by altering the subcellular trafficking of p53 during the cell cycle: the p53 remains in the cytoplasm of the cotransfected cells during a critical period in G1. This finding suggests a mechanism by which normal hematopoietic progenitors can survive and proliferate despite p53 expression and by which the inappropriate expression of bcl-2 and c-myc can cooperate in transformation.     

Long-term follow-up after transplantation of insulin-producing pancreatic islets into patients with Type 1 (insulin-dependent) diabetes mellitus

Summary Purified human islets and a kidney from the same donor were transplanted into four patients with Type 1 (insulin-dependent) diabetes mellitus. Two of the patients received additional islets that were isolated from multiple donors, cryopreserved, and stored in a tissue bank. The islets were embolized into the liver via the portal vein. Immunosuppression was induced with antilymphocyte globulin and maintained with azathioprine, prednisone and cyclosporine. In the first two patients, fasting serum C-peptide rose to levels of 0.5–2.0 ng/ml during the first 4–8 weeks and mixed meal feeding elicited increases to 2–3 ng/ml. C-peptide secretion persisted for 8 months, but at progressively lower levels and insulin therapy could not be withdrawn. In the next two patients who received cryopreserved islets in addition to fresh islets, serum C-peptide levels (fasting/post-meal) rose to 4–7 ng/ml and serum glucose was more stable, allowing withdrawal of insulin therapy after 69 days in one patient, and reduced insulin doses in the other. The insulin-independent patient has maintained normal fasting glucose, glycosylated haemoglobin, and oral glucose tolerance at 1 year following cessation of daily insulin therapy. Episodes of renal graft rejection occurred in three patients, including the insulin-independent patient. High-dose steroid therapy reversed the rejection in all instances, with apparent preservation of C-peptide secretion. These data show that transplantation of purified freshly-prepared and cryopreserved islets into Type 1 diabetic patients results in prolonged insulin secretion, and that sufficient function could be provided in one patient to sustain euglycaemia in the absence of insulin therapy at 1 year of follow-up.     

Fractional Flow Reserve Derived From Computed Tomographic Angiography in Patients With Multivessel CAD

Background

The functional SYNTAX score (FSS) has been shown to improve the discrimination for major adverse cardiac events compared with the anatomic SYNTAX score (SS) while reducing interobserver variability. However, evidence supporting the noninvasive FSS in patients with multivessel coronary artery disease (CAD) is scarce.

Interphase FISH to detect PBX1/E2A fusion resulting from the der(19)t(1;19)(q23;p13.3) or t(1;19)(q23;p13.3) in paediatric patients with acute lymphoblastic leukaemia

Approximately 6% of paediatric patients with precursor B-cell acute lymphoblastic leukaemia (B-ALL) harbour a rearrangement involving the gene regions of PBX1 (1q23) and E2A (19p13.3) which is visualized cytogenetically either as a der(19)t(1;19)(q23;p13.3) or the less common balanced t(1;19)(q23;p13.3). Unfortunately, no commercial dual-colour, double fusion fluorescence in situ hybridization (D-FISH) strategies are available to detect this recurrent anomaly. Therefore, we have created a D-FISH assay to detect these translocations and monitor minimal residual disease. This probe set was created using four bacterial artificial chromosomes (BACs) corresponding to the PBX1 gene region at 1q23 and four BACs corresponding to the E2A gene region at 19p13.3. We analysed 30 negative bone marrow controls and 20 diagnostic and post-treatment specimens from 13 paediatric B-ALL patients with a cytogenetically defined 1;19 translocation. Once unblinded, the results demonstrated that our D-FISH method effectively identified all diagnostic samples as abnormal and identified disease in four post-treatment samples that were previously considered to be normal by conventional cytogenetic analysis. The development of this FISH strategy for the detection of der(19)t(1;19)(q23;p13.3) and t(1;19)(q23;p13.3) proved to be an effective technique, allowing both the detection of disease in diagnostic samples and in post-treatment samples.     

Complete 360° circumferential gonioscopic optical coherence tomography imaging of the iridocorneal angle

Clinically, gonioscopy is used to provide en face views of the ocular angle. The angle has been imaged with optical coherence tomography (OCT) through the corneoscleral limbus but is currently unable to image the angle from within the ocular anterior chamber. We developed a novel gonioscopic OCT system that images the angle circumferentially from inside the eye through a custom, radially symmetric, gonioscopic contact lens. We present, to our knowledge, the first 360° circumferential volumes (two normal subjects, two subjects with pathology) of peripheral iris and iridocorneal angle structures obtained via an internal approach not typically available in the clinic.OCIS codes: (170.4500) Optical coherence tomography, (170.0110) Imaging systems, (170.4470) Ophthalmology     

Hand‐held Dynamometer Positioning Impacts Discomfort During Quadriceps Strength Testing: A Validity and Reliability Study

Purpose/Background

A belt‐stabilized hand‐held dynamometer (HHD) offers the ability to quantify quadriceps muscle strength in a clinical environment, but a limitation is participant discomfort at the interface between the HHD and the tibia. The purpose of this study was to quantify the level of discomfort associated with a modified belt‐stabilized HHD configuration compared to a standard belt‐stabilized configuration and an isokinetic dynamometer. The secondary purpose of this study was to determine the validity and reliability of a modified configuration used to measure quadriceps strength compared to the “gold‐standard” isokinetic dynamometer.

Methods

Twenty healthy participants (5 males, 15 females; age=24.7±2.2 years, height=171.1±8.8 cm, mass=72.0±18.7 kg) performed maximal knee extension isometric contractions during each of three testing conditions: isokinetic dynamometer, standard configuration with HHD placement on the tibia, and an alternative configuration with the HHD interfaced with the leg of a table. Discomfort was quantified using a Visual Analog Scale (VAS). Differences in discomfort and torque (N•m) associated with the testing positions were determined using Friedman test or repeated measures analysis of variance. Validity was quantified using Pearson correlations and within‐session intrarater reliability was determined using an intraclass correlation coefficient (ICC2,1) and associated confidence intervals (95% CI).

Results

The isokinetic dynamometer configuration resulted in the least discomfort (p< .01) and the modified configuration was more comfortable than the standard configuration (p= .003). There was a significant correlation between measures from the isokinetic dynamometer and the standard configuration (r=.87) and modified configuration (r=.93). Within‐session intrarater reliability was good for both the standard configuration (ICC_2,1=0.93) and modified configuration (ICC_2,1=0.93) conditions.

Conclusions

The use of the modified belt‐stabilized HHD configuration, where the HHD was interfaced with the leg of a table, offers a more comfortable alternative compared to the standard belt‐stabilized configuration to obtain isometric quadriceps strength measures in a clinical environment. This configuration is also a valid and reliable alternative to the “gold standard” isokinetic dynamometer when testing isometric quadriceps strength at 90° of knee flexion.

Level of Evidence

Diagnostic, Level 3     

Optimizing CT for the evaluation of vestibular aqueduct enlargement:Inter-rater reproducibility and predictive value of reformatted CT measurements

Enlarged vestibular aqueduct(EVA), the most frequent identifiable cause of congenital hearing loss, is evaluated with high-definition multidetector CT in the axial plane. Our purpose was to determine which reformatted CT measurements are most reproducible. Seven multiplanar reformatted images were created for each of the 64 temporal bones in patients with EVA. Intraclass correlation coefficients(ICC) were used to assess inter-observer variability, and both linear regression and ROC analyses were used to compare the measurements with severity of hearing loss, as assessed by pure tone audiometry. All seven measurements had excellent inter-observer variability, with average-measure ICC ranging from 0.92 to 0.98. There was no statistically significant correlation between the radiologic degree of aqueduct enlargement and severity of hearing loss using any of the seven measurements; ROC analyses revealed areas under the curves ranging from 0.57 to 0.73. Optimal accuracy was obtained with a threshold of 1.75 mm as measured at the aqueductal aperture in the P€oschl plane, with sensitivity of 0.75 and specificity of0.63. Although the radiologic measurement may not serve as a reliable tool for assessing severity of EVA, P€oschl plane reformatting has proven to be better than conventional axial acquisition plane for identifying patients with clinically significant hearing loss.     

Predictive value of electrocardiographic patterns in localizing left ventricular asynergy in coronary artery disease.

Transmural myocardial infarction by ECG (ECG-MI) was correlated with left ventricular asynergy by biplane left cineventriculography in 200 patients with coronary artery disease. The ability of individual ECG-MI patterns to predict and correctly localize asynergy was: anterior--98 per cent (43 of 44), inferior--82 per cent (36 of 44), true posterior--73 per cent (11 of 15). Of various combinations of criteria for true posterior ECG-MI, the pattern of an R wave and upright T wave in Lead V1 was most predictive of posterior asynergy--80 per cent (8 of 10). The LAO projection demonstrated a wall motion abnormality not appreciated in the RAO in 8 per cent (10 of 122) of cases of inferoposterior asynergy and enhanced assessment of asynergy in 30 per cent (36 of 122) of cases. It is concluded that: (1) ECG-MI has a high predictive accuracy for left ventricular asynergy, (2) an R-wave and upright T wave in Lead V1 is the best ECG predictor of posterior asynergy, and (3) the LAO projection makes an important contribution to the assessment of regional asynergy in coronary artery dieseas.     

Rate of FKS Mutations among Consecutive Candida Isolates Causing Bloodstream Infection

Precise FKS mutation rates among Candida species are undefined because studies have not systematically screened consecutive, disease-causing isolates. The Sensititre YeastOne (SYO) assay measures echinocandin MICs against Candida with less variability than reference broth microdilution methods. However, clinical breakpoint MICs may overstate caspofungin nonsusceptibility compared to other agents. Our objectives were to determine Candida FKS mutation rates by studying consecutive bloodstream isolates and to determine if discrepant susceptibility results were associated with FKS mutations. FKS hot spots were sequenced in echinocandin-intermediate and -resistant isolates and those from patients with breakthrough candidemia or ≥3 days of prior echinocandin exposure. Overall, 453 isolates from 384 patients underwent susceptibility testing; 16% were echinocandin intermediate or resistant. Intermediate susceptibility rates were higher for Candida glabrata than for other species (P < 0.0001) and higher for caspofungin than for other agents (P < 0.0001). Resistance rates were similar between agents. FKS mutations were detected in 5% of sequenced isolates and 2% of isolates overall. Corresponding rates among C. glabrata isolates were 8% and 4%, respectively. Among Candida albicans isolates, rates were 5% and <1%, respectively. Mutations occurred exclusively with prior echinocandin exposure and were not detected in other species. Isolates with discrepant susceptibility results did not harbor FKS mutations. Mutation rates among isolates resistant to ≥2, 1, and 0 agents were 75%, 13%, and 0%, respectively. In conclusion, FKS mutations were uncommon among non-C. glabrata species, even with prior echinocandin exposure. Discrepancies in echinocandin susceptibility by SYO testing were not driven by mutations and likely reflect imprecise caspofungin clinical breakpoints.