Citations

Cystic fibrosis is the most widespread hereditary disease among the white population caused by different mutations of the apical membrane ATP-binding cassette transporter cystic fibrosis transmembrane conductance regulator (CFTR). Its most common mutation, ΔF508, leads to nearly complete degradation via endoplasmic reticulum-associated degradation (ERAD). Elucidation of the quality control and degradation mechanisms might give rise to new therapeutic approaches to cure this disease. In the yeast Saccharomyces cerevisiae, a variety of components of the protein quality control and degradation system have been identified. Nearly all of these components share homology with mammalian counterparts. We therefore used yeast mutants defective in the ERAD system to identify new components that are involved in human CFTR quality control and degradation. We show the role of the lectin Htm1p in the degradation process of CFTR. Complementation of the HTM1 deficiency in yeast cells by the mammalian orthologue EDEM underlines the necessity of this lectin for CFTR degradation and highlights the similarity of quality control and ERAD in yeast and mammals. Furthermore, degradation of CFTR requires the ubiquitin protein ligases Der3p/Hrd1p and Doa10p as well as the cytosolic trimeric Cdc48p-Ufd1p-Npl4p complex. These proteins also were found to be necessary for ERAD of a mutated yeast “relative” of CFTR, Pdr5*p.

Concavity-compression is an important mechanism for keeping a joint centered despite a large range of motion. The purpose of this study was to explain how the results of a test measuring the joint intrinsic stability can be interpreted and related to joint architecture. As an example, the method was demonstrated for the glenohumeral joint although the versatility allows any articulating joint, whether natural or prosthetic, to be analyzed. The initial slope from the central point was relatively steep, indicating a large resistance to translation. The peak translation force occurred within the first 5 mm of displacement for the glenohumeral joint, indicating a high congruence between the humerus and glenoid surfaces. Stability ratio, calculated as the maximum translation force divided by the applied joint compressive force, makes it possible to compare the stabilizing effect under different compressive loads for different anatomical directions. In hanging arm position, the joint stability ratio ranged from 30.5% to 60.1%. Finally, the effective depth of the concavity and the maximum range of joint translation can be measured by completely dislocating the joint. For the glenohumeral specimen, the smallest glenoid concavity depth, 3.8 mm, occurred in the anterior direction. The joint translation limit was smallest in the anterior-posterior direction (28.0 mm). The methodology presented in this study will allow consistent testing parameters between different trials, easily allowing parametric studies to gain a more complete understanding of articular joints.

The supraspinatus tendon is frequently involved in rotator cuff tears. It has been suggested that joint position affects the structural mechanics of the tendon–bone complex. The purpose of this study was to determine regional variations in structural properties of the supraspinatus tendon in two glenohumeral positions. Supraspinatus tendons from 17 fresh frozen cadavers were divided into three strips of equal width and tested with a material-testing machine. The arm orientation was either in hanging position or 60 degrees glenohumeral abduction corresponding to 90 degrees arm abduction assuming 30 degrees scapular rotation. Tensile force, tendon elongation and failure mode were recorded. Overall, there was no significant difference in structural properties between hanging arm position and 60 degrees of glenohumeral abduction (p>0.05). However, the mean ultimate load (385 N, SD 56 N) and mean ultimate stress (14 MPa, SD 3 MPa) of the anterior tendon section with the arm in glenohumeral abduction were lower in 60 degrees abduction than in the hanging arm position (611 N, SD 276 N; 24 MPa, SD 10 MPa). In hanging arm position, the anterior tendon portion had a significantly greater ultimate load and stiffness than the middle and posterior portions (p<0.05). The regional variation in structural properties substantiates the clinical finding that rotator cuff ruptures easily extend posteriorly. Our study suggests that glenohumeral abduction reduces the failure strength of the supraspinatus tendon, specifically of its anterior portion. In our study, the maximum load of the anterior portion was substantially higher than predicted maximum loads transmitted physiologically through the entire tendon.

Background— Restenosis after percutaneous coronary intervention (PCI) is a major problem affecting 15% to 30% of patients after stent placement. No oral agent has shown a beneficial effect on restenosis or on associated major adverse cardiovascular events. In limited trials, the oral agent tranilast has been shown to decrease the frequency of angiographic restenosis after PCI. Methods and Results— In this double-blind, randomized, placebo-controlled trial of tranilast (300 and 450 mg BID for 1 or 3 months), 11 484 patients were enrolled. Enrollment and drug were initiated within 4 hours after successful PCI of at least 1 vessel. The primary end point was the first occurrence of death, myocardial infarction, or ischemia-driven target vessel revascularization within 9 months and was 15.8% in the placebo group and 15.5% to 16.1% in the tranilast groups ( P =0.77 to 0.81). Myocardial infarction was the only component of major adverse cardiovascular events to show some evidence of a reduction with tranilast (450 mg BID for 3 months): 1.1% versus 1.8% with placebo ( P =0.061 for intent-to-treat population). The primary reason for not completing treatment was ≥1 hepatic laboratory test abnormality (11.4% versus 0.2% with placebo, P <0.01). In the angiographic substudy composed of 2018 patients, minimal lumen diameter (MLD) was measured by quantitative coronary angiography. At follow-up, MLD was 1.76±0.77 mm in the placebo group, which was not different from MLD in the tranilast groups (1.72 to 1.78±0.76 to 80 mm, P =0.49 to 0.89). In a subset of these patients (n=1107), intravascular ultrasound was performed at follow-up. Plaque volume was not different between the placebo and tranilast groups (39.3 versus 37.5 to 46.1 mm 3 , respectively; P =0.16 to 0.72). Conclusions— Tranilast does not improve the quantitative measures of restenosis (angiographic and intravascular ultrasound) or its clinical sequelae.

Among the four subtypes of Hodgkin disease (HD), lymphocyte-predominant (LP) HD is now generally considered as a separate entity. The B cell nature of the typical Hodgkin and Reed–Sternberg (HRS) cells and their variants (L and H, lymphocytic and histiocytic cells) in LP HD has long been suspected, but the question of whether these cells represent a true tumor clone is unclear. We previously demonstrated clonal Ig gene rearrangements in one case of LP HD. In the present study, five cases of LP HD were analyzed by micromanipulation of single HRS cells from frozen tissue sections and DNA amplification of rearranged Ig heavy chain genes from those cells. Clonal V gene rearrangements harboring somatic mutations were detected in each case. In three cases ongoing somatic mutation was evident. This shows that HRS cells in LP HD are a clonal tumor population derived from germinal center B cells. The pattern of somatic mutation indicates that HRS cells in LP HD are selected for antibody expression. This, and the presence of ongoing mutation discriminates LP from classical HD.

Abstract We have recently described a modified model of progressive rapid ventricular pacing-induced heart failure which evolves over a period of 38 days. To further characterize left ventricular remodeling during the progression of heart failure, we assessed left ventricular geometry, wall stress, and atrial natriuretic peptide (ANP) gene expression and protein content during control conditions, asymptomatic left ventricular dysfunction, and overt congestive heart failure (CHF). Although asymptomatic left ventricular dysfunction was characterized by a significant increase in systolic and diastolic left ventricular dimension (+30% and +6%, respectively, P&lt;0.05 each) and a marked increase in left ventricular systolic wall stress (+68%, P&lt;0.01), left ventricular ANP gene expression was unchanged as compared to control. In contrast, strong left ventricular ANP gene expression (+449%, P&lt;0.05) was observed during overt CHF in the absence of further significant increases in left ventricular systolic wall stress. The onset of strong left ventricular ANP gene expression was associated with increased ANP content (+88%, P&lt;0.05) and left ventricular mass index (+13%, P&lt;0.05). In contrast, left atrial ANP gene expression and left ventricular diastolic wall stress increased progressively during asymptomatic left ventricular dysfunction (+39%, P=n.s. and +131%, P&lt;0.01) and overt CHF (+76% and +336% vs. control, P&lt;0.01 each). Progressive rapid ventricular pacing is associated with the induction of left ventricular ANP gene expression and protein synthesis exclusively during overt CHF. The current studies provide new insight into the temporal pattern of ANP-activation and the disparity between left ventricular systolic wall stress and ANP-activation in a large animal model of progressive CHF.