The binding and processing of monoclonal human IgG1 by cells of a human macrophage-like cell line (U937)

The goal of these experiments was to assess the relationship between the binding and processing of IgG by Fc-receptor-bearing cells. Cells of the U937 human macrophage-like cell line were incubated with 125I- labeled monomers, dimers, oligomers (composed of 2-4 IgG1 subunits), and HP (heavy polymers composed of 5 or more subunits per polymer) of monoclonal human IgG1 in vitro. Binding was assessed by spinning cells through a layer of phthalate oils. Internalization of IgG1 was assessed by quantitating residual binding to cells after surface-bound IgG was removed by a brief treatment with a solution containing 0.25 M acetic acid and 0.5 M sodium chloride. Catabolism was assessed by measuring the release of radioactive fragments of IgG1, which were not precipitated by 10% trichloroacetic acid. Unstimulated U937 bound about 10,000 molecules per cell of IgG1 monomer, with an equilibrium binding constant (Ka) of 5 X 10(8) M-1. After stimulation with a conditioned medium in vitro, binding per cell was increased 3-7--fold, and the Ka was decreased 2-4--fold. Both unstimulated and stimulated cells internalized and catabolized labeled IgG1 HP, but stimulated cells internalized and digested much more IgG1 HP per cell than unstimulated cells. Both monomers and dimers of IgG1 were internalized and degraded very slowly by stimulated cells, even though both preparations readily bound to cells. In contrast, oligomers and (to an even greater extent) IgG1 HP were internalized and degraded much more rapidly. Internalization of IgG1 HP was markedly inhibited by incubation at 4 degrees C, but not by incubation with a variety of metabolic inhibitors. Catabolism was inhibited by chloroquine and monensin (inhibitors of lysosomal acidification) and by cytochalasin (an inhibitor of microfilament polymerization). Binding to the surface of cells was not markedly inhibited by any agent tested. The capacity of cells to bind labeled IgG1 was markedly reduced by prior incubation in the presence of unlabeled IgG1. This reduction was in part due to the steric blockade of receptors caused by the avid, but reversible, binding of IgG1. In addition, IgG1 oligomers or HP (but not IgG1 monomers or dimers) also caused an irreversible reduction in the number of Fc receptors by a process analogous to receptor down-regulation, as observed in other receptor--ligand systems.     

Safety and Efficacy of Reteplase for the Treatment of Acute Arterial Occlusion: Complexity of Underlying Lesion Predicts Outcome

We evaluated both the safety and efficacy of reteplase for treatment of acute arterial occlusion as well as outcomes based on treatment of the underlying lesion. From November 2000 to February 2004, reteplase was used to treat arterial occlusions in 81 patients. Catheter-directed intrathrombus thrombolysis was performed with reteplase (0.5 units/hr) continuous infusion. Percutaneous mechanical thrombectomy (Angiojet) was performed in 61% (n = 50) of patients prior to thrombolysis. Unmasking of significant lesions resulted in endovascular intervention (39.5%), open surgical repair (24.6%), or both endovascular and surgical repair (9.8%) of the lesion. No lesion was found in 25.9% of patients. Major and minor complication rates as well as restoration of patency, limb salvage, and amputation-free survival were evaluated. Eighty-one patients received reteplase therapy (median = 10.3 +/- 5.3 units, 19.5 +/- 7.4 hr) followed by next-day arteriogram to assess thrombus removal. Technical success was achieved in 96.2% (n = 78) of cases. Kaplan-Meier life table analysis revealed overall primary patency rates of 76.3%, 60.1%, and 51.6%, at 1, 6, and 12 months, respectively. Overall amputation-free survival rates were 86.4%, 76.4%, and 69.7% at 1, 6, and 12 months, respectively. When subdivided into postlysis intervention, the lysis-only group achieved increased patency (p = 0.0143) and increased limb salvage (p = 0.0219) at 1 year compared to the lysis and endovascular intervention and the lysis and surgical groups. The 30-day complication rate was 17.3% (n = 14), with a major complication rate of 4.9% (n = 4) and a minor complication rate of 12.3% (n = 10). There were no intracranial hemorrhagic complications. Intra-arterial catheter-directed infusion of reteplase for acute lower extremity ischemia is safe and efficacious, as shown by the low risk of bleeding complications, high limb salvage rates, and low mortality rates in this study. The complexity of the lesion that is unmasked through thrombolytic therapy is a predictor of patency and limb salvage.     

Carotid angiography in contemporary vascular surgery practice

While most patients with carotid artery disease can safely undergo carotid endarterectomy based on duplex ultrasound alone, carotid angioplasty and stenting must, by its nature, be performed in conjunction with carotid arteriography. The techniques of carotid angiography are a necessary prerequisite to carotid intervention. The indications, technique, and results of carotid angiography in a contemporary vascular surgery practice are described.     

The isolation of thymidine-requiring Haemophilus influenzae from the sputum of chronic bronchitic patients receiving trimethoprim

Two different strains of thymidine-requiring Haemophilus influenzae (thy-) were isolated from the sputum of two patients with chronic obstructive airways disease, during therapy with oral trimethoprim for the treatment of an acute exacerbation of chronic bronchitis. Both patients responded clinically. Since trimethoprim levels in sputum and serum were very low the patients were probably non-compliant. Isolates of H. influenzae prototrophic for thymidine (thy+) were isolated from both patients in remission before, at the onset of exacerbation and also within two weeks of stopping therapy. Thy- and thy+ isolates from the same patient were of identical biotype but isolates from the two patients were of different biotypes. The minimum inhibitory concentration (MIC) of trimethoprim for the thy+ isolates obtained after therapy was significantly higher than the MIC for the isolates obtained at the onset of exacerbation. Thy- isolates were further investigated and a novel mechanism of low-level trimethoprim resistance is proposed.     

Alterations in rat renal glucose transport following in vivo use of Freund's adjuvant

Alterations in rat renal glucose transport following in vivo use of Freund's adjuvant were examined. Lewis-Brown Norway rats were placed in four separate injection groups: tubular basement membrane plus adjuvant [complete Freund's adjuvant (CFA) plus pertussis]; adjuvant (CFA plus pertussis); CFA only; or pertussis only. Renal handling of glucose was assessed 14 days after a single injection. No in vivo changes were noted. No histologic differences among groups were noted. However, brush border membrane vesicles prepared from animals in groups 1, 2, and 3 showed a marked decrease in glucose uptake. Further, Michaelis-Menten kinetics demonstrated a decrease in apparent Km and Vmax for glucose in groups 1, 2, and 3. CFA alone can cause a change in brush border membrane vesicle uptake of glucose. The pathogenic mechanism behind CFA-induced transport changes remains unclear. However, studies employing CFA cannot dismiss Freund's adjuvant as "inert" and must take into account functional changes created by CFA alone.     

First robotic endoscopic epicardial isolation of the pulmonary veins with microwave energy in a patient in chronic atrial fibrillation

The pulmonary veins have been demonstrated to play an important role in generating atrial fibrillation. We report the first successful endoscopic epicardial isolation of the pulmonary veins in a patient with permanent atrial fibrillation, along with a 1-year follow-up. The procedure consisted of making a conduction block around the pulmonary veins with a flexible microwave energy delivery probe. The probe was placed endoscopically on the left atrial epicardium with the aid of robotic instruments.     

Airway intervention in adult supraglottitis

Background The timing of aggressive airway intervention in adult epiglottitis is controversial. Aims To correlate Friedman’s staging of epiglottitis on admission with the airway interventions undertaken. Methods A retrospective study of 23 adult patients, mean age 51 years (range 29–81 years), who had been admitted with acute supraglottitis between March 1988 and December 2000 was undertaken. Results Three patients (13%) had airway interventions; two with tracheostomy and one with tracheal intubation. All were Friedman stage III and had rapid symptom progression during the 24 hours prior to admission. Three other stage III patients with symptom progression longer than 24 hours and all the remaining patients (stage II or less) were managed with observation and intravenous therapy. Conclusions Friedman originally advocated airway intervention in any patient stage II or worse, but this intubation threshold should probably be lowered to those patients with rapid-onset stage III (moderate respiratory distress, stridor, respiratory rate >30 per minute, pCO₂ >45mmHg) disease.