Effect of weight loss on inflammatory and endothelial markers and FMD using two low-fat diets.

OBJECTIVE: Cardiovascular disease is strongly associated with obesity and there is evidence that weight loss has positive effects on cardiovascular disease risk. The aims of this study were to compare meal replacements (MR) with a conventional low-fat diet as weight loss strategies and to examine the effect of weight loss on flow-mediated dilatation (FMD) and other markers of endothelial function in overweight Australians with raised triglycerides (TG) (> 2 mmol/l). RESEARCH METHODS: Subjects matched for age, gender, fasting plasma TG and body mass index were randomized to two low- fat high- carbohydrate weight loss strategies (both < 6000 kJ), one using MR and the other a structured eating plan, control (C). Subjects followed both diets for 3 months. In total, 55 subjects completed the study. FMD, pulse wave velocity and blood pressure (BP) were measured at baseline and at 3 months, as were fasting blood samples for lipids, glucose, insulin, C reactive protein (CRP) and endothelium-derived factors. RESULTS: Mean weight loss was 6.3 +/- 3.7 kg (6.0 +/- 4.2 vs 6.63 +/- 3.35 kg, MR vs C) with no difference between diet groups. TG, insulin, CRP, plasminogen activator inhibitor 1 (PAI-1) and soluble intracellular adhesion molecule-1 (sICAM1) fell after weight loss, but FMD did not change. Systolic BP fell by 8 mmHg and pulse wave velocity improved. DISCUSSION: In subjects with elevated TG, weight loss resulted in significant improvements in cardiovascular risk markers, particularly endothelium-derived factors (PAI-1 and sICAM1). However, FMD did not improve with weight loss.

Distribution of a bovine spongiform encephalopathy-derived agent over ion-exchange chromatography used in the preparation of concentrates of fibrinogen and factor VIII.

BACKGROUND AND OBJECTIVES: The risk of haemophiliacs contracting variant Creutzfeldt-Jakob disease (vCJD) via treatment with factor VIII concentrates is not known. Therefore, in order to determine the extent to which the vCJD agent might be removed during the preparation of factor VIII concentrate, the partitioning of a bovine spongiform encephalopathy (BSE)-derived agent was measured over the main purification step used to prepare the Scottish National Blood Transfusion Service high-purity factor VIII concentrate (Liberate). MATERIALS AND METHODS: Murine-passaged BSE (strain 301V), in the form of a microsomal fraction prepared from infected brain, was used to 'spike' a solution of factor VIII of intermediate purity. The 'spiked' starting material was subjected to solvent-detergent treatment and then to anion-exchange chromatography with Toyopearl DEAE-650M. All fractions were tested for 301V infectivity using a murine bioassay, including the procedures used to clean the ion-exchange media after use. RESULTS: BSE 301V infectivity was reduced by 2.9 log(10) in the fibrinogen fraction and by 2.7 log(10) in the factor VIII fraction. Over 99% of the added 301V infectivity remained bound to the ion-exchange column after elution of factor VIII. A large quantity of infectivity was subsequently removed by washing the ion-exchange media with 2 m NaCl. No further BSE 301V infectivity was detected in column eluates after treatment with 0.1 m NaOH or a second wash with 2 m NaCl. CONCLUSIONS: Results using a BSE-derived agent suggest that vCJD infectivity would be substantially removed by the ion-exchange process used in the preparation of fibrinogen and factor VIII concentrate. Although 301V infectivity remained bound to the ion-exchange matrix following elution of factor VIII, this appeared to be eliminated by the procedure used for cleaning the ion-exchange media after each use.

Studies on the removal of a bovine spongiform encephalopathy-derived agent by processes used in the manufacture of human immunoglobulin.

BACKGROUND AND OBJECTIVES: There is still uncertainty over how the agent of variant Creutzfeld-Jakob disease (vCJD) would partition during the manufacture of plasma derivatives. In this study, a BSE-derived agent was used as a vCJD model to determine the extent to which infectivity could be removed by selected steps used in the manufacture of intravenous immunoglobulin (IVIG). MATERIALS AND METHODS: Murine-passaged BSE (strain 301V), in the form of a microsomal fraction prepared from infected brain, was used to "spike" the starting material in three experiments. The partitioning of BSE infectivity was measured over Fraction I+III precipitation, borosilicate microfibre depth filtration and Seitz depth filtration, with these steps being examined individually and in series. RESULTS: Most 301V infectivity partitioned into Fraction I+III (log reduction 2.1). Infectivity remaining in Supernatant I+III was reduced by AP20 glass-fibre depth filtration (log reduction 0.6) and subsequently removed to below the limit of detection by Seitz KS80 depth filtration, giving an overall log reduction of > or = 2.9 for the three steps in series. By contrast, glass-fibre depth filtration gave a log reduction of 2.4 when challenged directly with "spiked" feedstock. Seitz KS80 depth filtration gave a log reduction of > or = 3.1 when challenged directly with 'spiked' feedstock and also removed residual infectivity to below the limit of detection when applied as the final step in series. CONCLUSIONS: Results using a BSE-derived agent suggest that vCJD infectivity should be substantially removed from immunoglobulin G (IgG) solutions by Fraction I+III precipitation and Seitz KS80 depth filtration. The three different process steps examined acted in a complementary manner to one another when operated in series. However, the data demonstrated that it would be inappropriate to add together the reduction factors that had been derived for each step in isolation.

Prions and blood products.

The transmission of Creutzfeldt-Jakob disease (CJD) by human pituitary-derived growth hormone has led to concerns that blood products might also provide a route for the iatrogenic transmission of CJD. A number of actions have been implemented by regulatory authorities to address such concerns, and numerous studies have been undertaken to determine whether or not there is a risk of CJD being transmitted in this manner. To date, no excess risk has been identified, leading to a growing consensus that there is little or no risk of long established forms of CJD being transmitted to recipients of blood products. This opinion does not extend to new variant CJD (vCJD) which is found predominantly in the UK and is believed to have resulted from the transmission of bovine spongiform encephalopathy (BSE) to humans. Unlike that of CJD, the prevalence of vCJD is not known. In addition, the detection of abnormal prion protein in the tonsils of vCJD-infected individuals has led to speculation that blood infectivity may be greater than in patients with CJD. A number of precautionary measures have been taken to address the possibility that vCJD may be transmissible by blood products; however, further scientific advances are needed to enable this risk to be defined. A suitable screening test is required to identify any infected blood donors, particularly where cellular blood components are being derived from populations believed to be at risk from BSE infection. Recent experimental data suggest that process operations used in the manufacture of plasma products may be capable of removing prion agents to a significant extent. However, further work is required to confirm these observations and to determine whether or not all potential vCJD infectivity would be removed by these means.

Studies on the removal of abnormal prion protein by processes used in the manufacture of human plasma products.

BACKGROUND AND OBJECTIVES: To identify if any process steps used in plasma fractionation may have a capability of removing agents of human transmissible spongiform encephalopathy (TSE). MATERIALS AND METHODS: Sixteen fractionation steps were investigated separately by adding a preparation of hamster adapted scrapie 263K to the starting material at each process step and determining the distribution into resultant fractions of protease-K-resistant (abnormal) prion protein by Western blot analysis. RESULTS: A number of process operations were found to remove abnormal prion protein to the limit of detection of the assay. These were cold ethanol precipitation of fraction IV (log reduction, LR, >/=3.0) and a depth filtration (LR >/=4.9) in the albumin process; cold ethanol fraction I+III precipitation (LR >/=3.7) and a depth filtration (LR >/=2.8) in the immunoglobulin processes and adsorption with DEAE-Toyopearl 650M ion exchanger (LR >/=3.5) in the fibrinogen process. In addition, a substantial degree of removal of abnormal prion protein was observed across DEAE-Toyopearl 650M ion exchange (LR = 3.1) used in the preparation of factor-VIII concentrate; DEAE-cellulose ion exchange (LR = 3.0) and DEAE-sepharose ion exchange (LR = 3.0) used in the preparation of factor-IX concentrates and S-sepharose ion exchange (LR = 2.9) used in the preparation of thrombin. CONCLUSIONS: Plasma fractionation processes used in the manufacture of albumin, immunoglobulins, factor-VIII concentrate, factor-IX concentrates, fibrinogen and thrombin all contain steps which may be capable of removing causative agents of human TSEs.

Interactions between electronic article surveillance systems and implantable cardioverter-defibrillators.

BACKGROUND: In patients with implantable cardioverter-defibrillators (ICDs). inappropriate shocks have been reported with exposure to electronic article surveillance systems. The risk to patients with ICDs of walking through or lingering near surveillance systems requires further investigation. METHODS AND RESULTS: We evaluated the response in ICD function in 170 subjects during a 10- to 15-second midgate walk-through of and during extreme (2 minutes within 6 in of the gate) exposure to 3 common article surveillance systems. Complete testing was done in 169 subjects. During a 10- to 15-second (very slow) walk-through of the 3 surveillance systems, no interactions were observed that would negatively affect ICD function. During extreme exposure (169 subjects) and during extreme exposure and pacing via the ICD (126 subjects), interactions between the ICD and the article surveillance systems were observed in 19 subjects. In 7 subjects, this interaction was clinically relevant and would have likely (3 subjects) and possibly (4 subjects) resulted in ICD shocks. In 12 subjects, the interaction was minor. CONCLUSIONS: It is safe for a patient with an ICD to walk through electronic article surveillance systems. Lingering in a surveillance system may result in an inappropriate ICD shock.

Assessment of the potential of plasma fractionation processes to remove causative agents of transmissible spongiform encephalopathy.

Although there is no evidence that classical CJD (cCJD) can be transmitted by human blood or blood products in clinical practice, uncertainties surrounding new variant CJD (nvCJD) have led to the safety of plasma products derived from UK donors being questioned. To better define whether or not there is a risk of nvCJD being transmitted it is necessary to determine how the causative agent would partition across the separations processes used in the preparation of plasma products. The abnormal prion protein which is associated with transmissible spongiform encephalopathies (TSEs), such as CJD, has a low solubility, a high tendency to form aggregates and adheres to surfaces readily. If the physicochemical properties of the agent of nvCJD are similar to those of abnormal prion protein then nvCJD may be removed by precipitation and adsorption technologies used in plasma fractionation. Available data on the removal of TSE agents by such bioprocess technologies have been used to estimate the potential degree of reduction expected from each step in the plasma fractionation processes used by the SNBTS. The overall process reduction factors estimated are: 10(13) (albumin). 10(9) (immunoglobulins), 10(7) (factor IX, thrombin), 10(5) (fibrinogen), 10(4) (factor VIII) and 10(3) (factor II, IX and X); however, it will be necessary to establish the accuracy of these estimates by practical validation studies.

Intracoronary infusion of dilute ethanol for control of ventricular rate in patients with atrial fibrillation.

The effects of selective infusion of 25% ethanol into the AV nodal artery was assessed in 11 patients with atrial fibrillation and uncontrollably rapid ventricular response rates. The primary study objective was to achieve permanent modification of AV nodal function and control ventricular rate without drug therapy and without causing permanent complete AV block. "Clinical success" was defined as drug-free rate control by either AV nodal modification or the production of complete AV block. Selective catheterization and ethanol infusion into the AV nodal artery could be performed in nine patients. Intracoronary ethanol infusion acutely caused second- or third-degree AV nodal block in seven patients and an increase in AV nodal refractory period and Wenckebach cycle length in two patients. Acute occlusion of the AV nodal artery or infarction of nontarget myocardium was not observed. During follow-up of 22.2 +/- 2.2 months the primary study objective was attained in only four of nine patients treated, yielding an efficacy of 44%. However, the "clinical success" rate was 78%. The acute effects of ethanol on AV conduction did not predict the chronic effects. Selective intracoronary infusion of dilute ethanol to control the ventricular rate in atrial fibrillation should be considered when radiofrequency ablation has been unsuccessful. This method of chemical ablation is as effective and probably safer than rapid administration of 96% ethanol.

Subxiphoid approach for implantable cardioverter defibrillator in patients with previous coronary bypass surgery.

From February 1988 until August 1991, 28 patients with prior coronary artery bypass grafting (CABG) received implantable cardioverter defibrillator (ICD) therapy via a subxiphoid approach. Only one patient required conversion to a median sternotomy incision. The mean defibrillation threshold was 11.9 +/- 4.4 J. The mean R wave was 8.2 +/- 3.7 mV. One perioperative death occurred due to heart failure (mortality rate 1/28 [3.50%]). No patient required reexploration for bleeding. The subxiphoid method for ICD electrode implantation is safe and reliable in patients with prior CABG surgery.

Combined coronary artery bypass grafting and bilateral renal revascularization for unstable angina and impeding renal failure.

The purpose of our article is to describe a patient with severe hypertension and moderate renal insufficiency, unstable angina, and a 6 cm abdominal aortic aneurysm. A previous aortogram had demonstrated severe bilateral renal artery stenoses. Cardiac catheterization demonstrated severe coronary disease. After cardiac catheterization acute renal failure and pulmonary edema requiring dialysis developed in the patient. In addition, evidence of impending myocardial necrosis developed. Because of the critical nature of the myocardial and renal ischemia it was necessary to perform combined myocardial and renal revascularization rather than staged procedures. At the time of coronary artery bypass grafting, a vein graft was anastomosed to the right coronary artery vein graft and tunneled through the diaphragm into the abdomen to revascularize both renal arteries. After surgery renal function gradually improved, and no further dialysis was required. The abdominal aortic aneurysm was repaired at a subsequent operation. At 2-year follow-up all grafts remained patent. The serum creatinine is 1.2 mg/dl. Although most patients with combined coronary artery disease and renal artery disease can be treated with staged operations, our procedure may be of value in patients in whom staged procedure are not feasible and in whom the infrarenal aorta is severely diseased or aneurysmal.

Studies on the stability of VIII:C during the manufacture of a factor VIII concentrate for clinical use.

The stability of VIII:C was investigated by monitoring samples taken at different points from a routine process for the manufacture of factor VIII concentrate and by examining the stabilising influence of a number of product formulations. Loss of VIII:C over process-finishing procedures (formulation, 0.22 micron filtration, dispensing) was associated with a citrate-induced inactivation which could be prevented by controlling the ionised calcium concentration of the solution. These results were obtained using a one-stage clotting assay but were not observed using a two-stage assay. No evidence for activation was found in vitro (e.g. by FPA generation and VIII:C stability) and the yield increase suggested by the one-stage assay was supported by results from a controlled clinical evaluation.