The anti-CD14 antibody IC14 suppresses ex vivo endotoxin stimulated tumor necrosis factor-alpha in patients with chronic heart failure

BACKGROUND: Activation of the endotoxin (LPS) receptor, CD14, leads to tumor necrosis factor-alpha (TNF) production. Plasma LPS activity is elevated in patients with severe chronic heart failure (CHF). An anti-CD14 antibody, IC14, blocks TNF production in healthy volunteers. It is not known whether IC14 prevents TNF production in CHF patients. METHODS AND RESULTS: Blood from 20 CHF patients (age 64+/-2.1 years, NYHA class 2.2+/-0.1, LVEF 27+/-3%, mean+/-SEM) was pre-incubated with 0.5, 1.0, 5.0, 10 and 50 microg/mL IC14 for 1 h followed by incubation with 1 or 10 ng/mL LPS for 6 h. Fourteen subjects served as controls (58+/-2.4 years). LPS-stimulated TNF release was 76% and 60% greater at 1 and 10 ng/mL LPS, respectively, in CHF patients versus controls (p=0.07 and p=0.008). IC14 at concentrations of 5.0, 10 and 50 microg/mL substantially reduced TNF production in response to stimulation with LPS (all p<0.05). CD14 receptor density was similar in patients and controls. In controls, but not in CHF patients, there was a positive correlation between CD14 receptor density and TNF production (r=0.61, p=0.03). CONCLUSION: IC14 suppresses LPS-stimulated whole blood TNF production in patients with CHF and in normal subjects and therefore may represent a novel therapeutic strategy for CHF patients with systemic immune activation.     

Deletion of the distal COOH-terminus of the A2B adenosine receptor switches internalization to an arrestin- and clathrin-independent pathway and inhibits recycling

Background and purpose:

We have investigated the effect of deletions of a postsynaptic density, disc large and zo-1 protein (PDZ) motif at the end of the COOH-terminus of the rat A_2B adenosine receptor on intracellular trafficking following long-term exposure to the agonist 5′-(N-ethylcarboxamido)-adenosine.

Experimental approach:

The trafficking of the wild type A_2B adenosine receptor and deletion mutants expressed in Chinese hamster ovary cells was studied using an enzyme-linked immunosorbent assay in combination with immunofluorescence microscopy.

Key results:

The wild type A_2B adenosine receptor and deletion mutants were all extensively internalized following prolonged treatment with NECA. The intracellular compartment through which the Gln³²⁵-stop receptor mutant, which lacks the Type II PDZ motif found in the wild type receptor initially trafficked was not the same as the wild type receptor. Expression of dominant negative mutants of arrestin-2, dynamin or Eps-15 inhibited internalization of wild type and Leu³³⁰-stop receptors, whereas only dominant negative mutant dynamin inhibited agonist-induced internalization of Gln³²⁵-stop, Ser³²⁶-stop and Phe³²⁸-stop receptors. Following internalization, the wild type A_2B adenosine receptor recycled rapidly to the cell surface, whereas the Gln³²⁵-stop receptor did not recycle.

Conclusions and implications:

Deletion of the COOH-terminus of the A_2B adenosine receptor beyond Leu³³⁰ switches internalization from an arrestin- and clathrin-dependent pathway to one that is dynamin dependent but arrestin and clathrin independent. The presence of a Type II PDZ motif appears to be essential for arrestin- and clathrin-dependent internalization, as well as recycling of the A_2B adenosine receptor following prolonged agonist addition.     

Ceramic-on-ceramic vs ceramic-on-polyethylene,a comparative study with 10-year follow-up

BACKGROUNDIn press-fit total hip arthroplasty (THA) ceramic-on-ceramic (CoC) bearings are a potential for overcoming the wear that is seen in ceramic-on-polyethylene (CoPE) bearings, and can lead to wear-induced osteolysis, resulting in loosening of the implant. However, CoC bearings show disadvantages as well, such as squeaking sounds and being more fragile, which can cause ceramic head or liner fracture. Because comparative long-term studies are limited, the objective of this study was to determine the long-term difference in wear, identify potential predictive factors for wear, investigate radiological findings such as osteolysis, and evaluate clinical functioning and complications between these bearings.AIMTo determine 10-year differences in wear, predictive factors for wear, and investigate radiological findings and clinical functioning between CoC and CoPE.METHODSThis observational prospective single-center cohort study with a 10-year follow-up includes a documented series of elective THAs. Primary outcome was wear measured by anteroposterior (AP) radiographs. Secondary outcomes were potential predictive factors for wear, complications during follow-up, Harris hip score (HHS), and radiological findings such as presence of radiolucency, osteolysis, atrophy, and hypertrophy around the cup. Due to the absence of wear in the CoC group, stratified analysis to identify risk factors for wear was only performed in the CoPE group by use of univariate linear regression analysis. HHS was expressed as a change from baseline and the association with bearing type was assessed by use of multivariate linear regression analysis, adjusted for potential confounders.RESULTSA total of 17 CoPE (63.0%) and 25 CoC (73.5%) cases were available for follow-up and showed a linear wear of respectively 0.130 mm/year (range 0.010; 0.350) and 0.000 mm/year (range 0.000; 0.005), which was significant (P < 0.001) between both groups. Wear always occurred in the cranial direction. Cup inclination was the only predictive factor for polyethylene (PE) wear. No dislocations, ceramic head, or liner fractures were seen. The HHS showed a mean change from baseline of 37.1 points (SD 18.5) in the CoPE group and 43.9 (SD 17.0) in the CoC group. This crude difference of 6.8 (range -5.2; 18.7) in favor of the CoC group was not significant (P = 0.26) and was not significant when adjusted for age, gender, and diagnosis either (P = 0.99). No significant differences in complications and radiological findings were seen between groups. CONCLUSIONCoC bearing shows lower wear rates compared to CoPE at 10-year follow-up with cup inclination as a predictive factor for wear and no differences in complications, HHS, and radiological findings.     

Ventilatory and diffusion abnormalities in long-term survivors after orthotopic heart transplantation.

OBJECTIVE: To investigate the long-term development of pulmonary diffusion abnormalities after orthotopic heart transplantation (oHT). DESIGN: Retrospective analysis of pulmonary function test results of different patient groups at different time intervals after oHT was performed. PATIENTS: This investigation included 642 patients who had undergone oHT for chronic heart failure. Patients were grouped according to the time elapsed after transplantation (group 1: n = 164; age, 47 +/- 14 years; days after oHT, 324 +/- 101; group 2: n = 100; age, 48 +/- 15 years; days after oHT, 723 +/- 104; group 3: n = 106; age, 52 +/- 12 years; days after oHT, 1,092 +/- 98; group 4: n = 84; age, 51 +/- 13 years; days after oHT, 1,442 +/- 99; group 5: n = 61; age, 50 +/- 14 years; days after oHT, 1,819 +/- 105; group 6: n = 101; age, 53 +/- 12 years; days after oHT, 2,463 +/- 303; and group 7: n = 26; age, 54 +/- 14 years; days after oHT, 3,478 +/- 246). In 56 (group 8) of the 642 patients, follow-up measurements were performed with tests before and at two time points after oHT (6.5 +/- 1.7 and 12.5 +/- 9.3 months). RESULTS: Of all patients, 39% showed restrictive and obstructive abnormalities with no differences between the groups. No significant differences in lung transfer factor for carbon monoxide (DLCO) were observed (61.2 vs 63.7 vs 65.5 vs 65.6 vs 64.5 vs 65.7 vs 67.6% predicted). Differences in transfer coefficient for carbon monoxide (Kco) were significant between group 1 and 4 (58.7 vs 64.1% predicted), and group 1 and 6 (58.7 vs 63.4% predicted). No differences occurred in the rate with which patients exhibited pathologic abnormalities for DLCO and KCO. After oHT, a marked reduction in diffusion capacity occurred in group 8. On follow-up, these measurements were only slightly restored in terms of the predicted DLCO percentage. No such improvement was observed in KCO or in the rate of pathologic changes for both DLCO and KCO. We conclude, therefore, that the impairment of diffusion does not improve even after a significant period has passed after the oHT. Whether this has any effect on symptoms and/or the prognosis for these patients is extremely unclear.