Purification and quantification of recombinant Epstein-Barr viral glycoproteins gp350/220 from Chinese hamster ovary cells

Truncated Epstein-Barr virus (EBV) membrane antigen gp350/220 (EBV-MA) lacking the membrane anchor was expressed and secreted into the medium of recombinant Chinese hamster ovary cells that had been cultured in Plasmapur hollow-fibre modules using defined serum-free medium. The EBV-MA in the medium was concentrated by 70% (w/v) ammonium sulphate precipitation and subsequently purified by immunoaffinity chromatography using an anti-EBV-MA (EBV.0T6) monoclonal antibody (mAb) column. Adsorbed antigen was eluted with 3 M MgCl2 in phosphate-buffered saline, concentrated by Mono Q anion-exchange chromatography and analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis, silver staining and Western blotting using EBV-positive serum and anti-EBV-MA specific mAbs. Monospecific polyclonal rabbit antibodies against the purified EBV-MA were raised and purified by protein G affinity chromatography. For the measurement of EBV-MA antigen levels a sandwich enzyme-linked immunosorbent assay using rabbit polyclonal antibodies and a horseradish peroxidase-conjugated anti-MA mAb was developed having a detection level of 10 ng/ml

Optimal nutrition during the period of mechanical ventilation decreases mortality in critically ill, long-term acute female patients: a prospective observational cohort study

ABSTRACT: INTRODUCTION: Optimal nutrition for intensive care patients has been proposed to be the provision of energy as determined by indirect calorimetry, and protein provision of at least 1.2 grams/kg pre-admission weight per day. The evidence supporting these nutritional goals is based on surrogate outcomes and is not yet substantiated by patient oriented, clinically meaningful endpoints. In the present study we evaluated the effects of achieving optimal nutrition in intensive care unit (ICU) patients during their period of mechanical ventilation on mortality. METHODS: Prospective observational cohort study in a mixed medical-surgical, 28-bed intensive care unit in an academic hospital. 243 sequential mixed medical-surgical patients were enrolled on day 3 to 5 after admission if they had an expected stay of at least another 5 to 7 days. They underwent indirect calorimetry as part of routine care. Nutrition was guided by the result of indirect calorimetry and we aimed to provide at least 1.2 grams of protein/kg/day. Cumulative balances were calculated for the period of mechanical ventilation. Outcome parameters were ICU, 28-day and hospital mortality. RESULTS: In women, when corrected for weight, height, Apache II score, diagnosis category, and hyperglycaemic index, patients who reached their nutritional goals compared to those who did not, showed a hazard ratio (HR) of 0.199 for ICU mortality (confidence interval [CI] 0.048 - 0.831; P = 0.027), a HR of 0.079 for 28 day mortality (CI 0.013 - 0.467; P = 0.005) and a HR of 0.328 for hospital mortality (CI 0.113 - 0.952; P = 0.04). Achievement of energy goals whilst not reaching protein goals, did not affect ICU mortality; the HR for 28-day mortality was 0.120 (CI 0.027 - 0.528; P = 0.005) and 0.318 for hospital mortality (CI 0.107 - 0.945; P=0.039). No difference in outcome related to optimal feeding was found for men. CONCLUSIONS: Optimal nutritional therapy improves ICU, 28-day and hospital survival in female ICU patients. Female patients reaching both energy and protein goals have better outcomes than those reaching only the energy goal. In the present study men did not benefit from optimal nutritio

Cervical necrotizing fasciitis with thoracic extension after total laryngectomy

Cervical necrotizing fasciitis (CNF) with thoracic extension is rare. It has never been reported in laryngectomized patients. A case of fatal CNF in a laryngectomized patient equipped with a voice prosthesis is presented. Diagnosis and treatment are discussed. CNF with thoracic extension was diagnosed on clinical picture, computed tomography (CT) and biopsies were taken just above the tracheostoma. Antibiotic treatment was started and extensive debridement of the affected tissues performed. A minor extension to the left pleura was considered irresectable. Irradical debridement and the impossibility of administering hyperbaric oxygen therapy caused death within two day after presentation. CNF is a rare disease and to our knowledge, has never been reported after total laryngectomy. This case emphasizes the need for early antibiotic treatment and radical surgical resection of the affected tissues

An algorithm for balanced protein/energy provision in critically ill mechanically ventilated patients

Background & aims: Optimal nutritional therapy for energy and protein in critically ill, mechanically ventilated patients can be defined as: providing calories matched with the measured energy expenditure and delivery of protein in an amount of 1.2-1.5 g/kg pre-admission weight/day. Several enteral nutritional products are available with different energy/protein proportions. We developed an algorithm to choose the nutritional formula that combines optimal energy and protein supply for individual patients. Methods: The energy and protein values of three nutritional formulas were used together with an aimed provision of 1.2-1.5 g protein/kg/day to construct a nomogram. From that, an algorithm followed, which was tested retrospectively in 203 mechanically ventilated patients with a normal BMI and known values for energy expenditure and weight. Results: In the nomogram cut-off points for energy/weight ratios were: 19.0-23.8 for a normal energy/high protein formula, 23.8-29.8 for a high energy/high protein formula and 30-37.5 for the normal energy/normal protein formula. The algorithm uses energy expenditure/body weight ratio of the patients to choose one of the three formulas. This resulted in an adequate provision of protein in 93% of the patients. Conclusion: The algorithm leads to provision of adequate amounts of protein and energy in the majority of critically ill, mechanically ventilated patients

Harvesting, logistics and upgrading of herbaceous biomass from verges and natural areas for use in thermal conversion : TKI BBE Park Cuijk

The main goal of this project was to analyse if it is technically possible, environmentally favourable and economically profitable to use herbaceous biomass from verges (roadsides) and natural areas as raw material for combustion in the Cuijk facility of Essent for the production of CHP (combined heat and power)