Isolation of the factor VIII-von Willebrand factor complex directly from plasma by gel filtration

A high capacity gel filtration system was developed with the purpose of isolating factor VIII (FVIII) and von Willebrand factor (vWF) directly from plasma in significantly higher yields than obtained by cryoprecipitation, the technique most commonly used to recover FVIII-vWF from human plasma. After laboratory-scale gel filtration of plasma, a FVIII-containing fraction was collected containing about 90% of FVIII in the applied plasma and with almost tenfold higher purity than that obtained by cryoprecipitation. The gel filtration step has been scaled up for use as the initial step in the manufacturing process for a FVIII preparation (Nordiate).     

In vivo purging with high-dose cytarabine followed by high-dose chemoradiotherapy and reinfusion of unpurged bone marrow for adult acute myelogenous leukemia in first complete remission

PURPOSE: To evaluate in a prospective study the efficacy of autologous bone marrow transplantation (BMT) in adult patients with acute myelogenous leukemia (AML) in first remission, using a single course of high-dose Cytarabine (HD Ara-C) consolidation therapy as in vivo purging. PATIENTS AND METHODS: Sixty consecutive adult patients with AML in first complete remission (CR) were treated with HD Ara-C consolidation therapy as a method of in vivo purging before marrow collection. High-dose therapy consisted of fractionated total-body irradiation (FTBI) 12 Gy, intravenous etoposide 60 mg/kg, and cyclophosphamide 75 mg/kg, followed by reinfusion of cryopreserved marrow. RESULTS: Sixty patients underwent consolidation treatment with HD Ara-C with the intent to treat with autologous BMT. Sixteen patients were unable to proceed to autologous BMT (10 patients relapsed, one died of sepsis, one developed cerebellar toxicity, two had inadequate blood counts, and two refused). Forty-four patients underwent autologous BMT and have a median follow-up time of 37 months (range, 14.7 to 68.7) for patients who are alive with no relapse. The cumulative probability of disease-free survival (DFS) at 24 months in the intent-to-treat group is 49% (95% confidence interval [CI], 37% to 62%) and in those who actually underwent autologous BMT is 61% (95% CI, 46% to 74%). The probability of relapse was 44% (95% CI, 31% to 58%) and 33% (95% CI, 20% to 49%) for the intent-to-treat and autologous BMT patients, respectively. CONCLUSION: This approach offers a relatively high DFS rate to adult patients with AML in first CR. The results of this study are similar to those achieved with allogeneic BMT.     

Physiologo-biochemical characteristics if Gluconobacter oxydans and prospects for its use in biotechnology and biosensor systems (review)

Gluconobacter oxydans possesses a unique organization of metabolic systems, which are characterized by reduction of major dissimilation pathways, surface localization of main oxidative enzymes responsible for partial oxidation of carbon substrates, high performance of electron-transport chains, and accumulation of partially oxidized metabolites in the medium. These features allow us to use the cells of these microorganisms in biotechnology for production of several food products and medicines. The use of G. oxydans in biosensors for estimation of concentrations of sugars, aldoses and polyalcohols is promising. Physiological and biochemical features of these microorganisms enabling their use in biotechnology and receptor elements of biosensors are reviewed.     

Identification and characterization of human metabolites of CAI [5-amino-1-1(4''-chlorobenzoyl-3,5-dichlorobenzyl)-1,2,3-triazole- 4-carboxamide)

BACKGROUND: Bronchoscopic bronchoalveolar lavage in children to investigate bronchial disorders such as asthma has both ethical and procedural difficulties. OBJECTIVE: The aim of this study was to establish a standardized non-bronchoscopic method to perform bronchoalveolar lavage in children attending for elective surgery to obtain normal cellular data. METHODS: Bronchoalveolar lavage was performed on normal children (n = 55) by infusing saline (20 mL) through an 8 FG suction catheter passed after endotracheal intubation. Oxygen saturation, heart and respiratory rate were monitored during the bronchoalveolar lavage procedure. Cellular analysis and total protein estimation of the lavage fluid were performed. Epithelial lining fluid volume was calculated (n = 15) using the urea dilution method. RESULTS: The procedure was well tolerated by all children. Total cell count and differential cell count for children (macrophages 70.8 +/- 2.3%, lymphocytes 3.8 +/- 0.6%, neutrophils 5.7 +/- 1.0%, eosinophils 0.14 +/- 0.03%, epithelial cells 19.6 +/- 2.1%, mast cells 0.21 +/- 0.02%) were similar to those reported for adults. Age and sex comparisons revealed no differences between groups. The mean total protein recovered in the cell free supernatant was 49.72 +/- 4.29 mg/L and epithelial lining fluid volume was 0.82 +/- 0.11% of return lavageate. CONCLUSION: This method allows bronchoalveolar lavage to be performed safely and quickly on children attending for routine elective surgery. Using this method and taking the 'window of opportunity' of elective surgery, the presence or absence of airway inflammation could be studied in children with various patterns of asthma during relatively asymptomatic periods.     

Effect of intracolonic benzalkonium chloride on trinitrobenzene sulphonic acid-induced colitis in the rat

BACKGROUND: We investigated the effects of benzalkonium chloride (BAC) on trinitrobenzene sulphonic acid (TNBS)-induced colitis in rats. METHODS: TNBS was administered intrarectally before and/or after BAC treatment. In the first study, the effects of treatment with BAC 6, 12 or 24 h after TNBS were examined. In the second study, animals were treated with BAC before, after or before and after TNBS, and were examined 7 days later. The severity of colitis was assessed by macroscopic and histological scoring of the colonic damage and by determination of colonic myeloperoxidase (MPO) activity. Macrophages and CD4+ and CD8+ T cells were examined by immunohistochemistry. RESULTS: When BAC was instilled into the colon 6, 12 or 24 h after TNBS, weight loss and macroscopic and histological features of the colon were similar to that of controls (TNBS alone). In contrast, MPO activity was significantly reduced in all three groups post-treated with BAC. In the groups examined 7 days after TNBS treatment, rats post-treated with BAC exhibited increased weight gain and significantly reduced macroscopic damage and MPO activity compared to the TNBS control group. Rats pre-treated with BAC exhibited less macroscopic damage of the colon than rats receiving only TNBS, but histological damage, MPO and weight gain were unchanged from TNBS controls. Immunohistochemistry revealed that BAC pre-treatment increased the numbers of macrophages and T cells in the colon. After TNBS treatment, macrophage accumulation was evident in the colon, but T cells were scarce. However, these cells were preserved or enhanced in the colonic mucosa in TNBS-treated rats that had been pre-treated with BAC. CONCLUSIONS: Treatment with BAC, particularly after induction of colitis, produces a significant reduction in the severity of tissue injury and inflammation through mechanisms that are not fully understood.     

Optimisation of solar photovoltaic (PV) parameters using meta-heuristics

Microsystem Technologies - This paper presents a critical analysis of the meta-heuristic techniques used in various researches on the optimisation of photovoltaic (PV) parameters, which involves...     

Impact of agriculture on soil consumption of atmospheric CH4 and a comparison of CH4 and N2O flux in subarctic, temperate and tropical grasslands

Increasing concentrations of methane (CH₄) in the atmosphere are projected to account for about 25% of the net radiative forcing. Biospheric emissions of CH₄ to the atmosphere total approximately 400 Tg C y^-1. An estimated 300 Tg of CH₄-C y^-1 is oxidized in the atmosphere by hydroxyl radicals while about 40 Tg y^-1 remains in the atmosphere. Approximately 40 Tg y^-1 of the atmospheric burden is oxidized in aerobic soils. Research efforts during the past several years have focused on quantifying CH₄ sources while relatively less effort has been directed toward quantifying and understanding the soil sink for atmospheric CH₄. Recent research has demonstrated that land use change, including agricultural use of native forest and grassland systems has decreased the soil sink for atmospheric methane. Some agricultural systems consume atmospheric CH₄ at rates less than 10% of those found in comparable undisturbed soils. While it has been necessary to change land use practices over the past centuries to meet the required production of food and fiber, we need to recognize and account for impacts of land use change on the biogeochemical nutrient cycles in the biosphere. Changes that have ensued in these cycles have and will impact the atmospheric concentrations of CH₄ and N₂O. Since CH₄ and N₂O production and consumption are accomplished by a variety of soil microorganisms, the influence of changing agricultural, forest, and, demographic patterns has been large. Existing management and technological practices may already exist to limit the effect of land use change and agriculture on trace gas fluxes. It is therefore important to understand how management and land use affect trace gas fluxes and to observe the effect of new technology on them. This paper describes the role of aerobic soils in the global CH₄ budget and the impact of agriculture on this soil CH₄ sink. Examples from field studies made across subarctic, temperate and tropical climate gradients in grasslands are used to demonstrate the influence of nutrient cycle perturbations on the soil consumption of atmospheric CH₄ and in increased N₂O emissions. This revised version was published online in August 2006 with corrections to the Cover Date.