Le nuove organizzazioni Open Source: ecosistemi di business in azione

2008-04-17Sardegna Ricerche, Edificio 2, Località Piscinamanna 09010 Pula (CA) - ItaliaPAAL 2008 - Pubblica Amministrazione Aperta e Libera: dalle tecnologie aperte alla libera circolazione dei contenuti digital

Total Protein Determination Using Micro Plate Assay

One of the largest challenges that food manufactures are facing today is the management of food allergens. Allergenic protein in trace amounts, part per million concentrations, will trigger a reaction in some individuals. Food manufacturers need to prevent allergen cross-contamination by performing adequate sanitation after production of an allergenic containing food. Allergen detection kits are used to determine if sufficient protein was removed from the equipment surfaces during sanitation. One kit on the market is the 3M™ Clean- Trace™ Surface Protein (Allergen) swab tests. The test qualitatively detects the presence of protein based on the biuret reaction and will yield a purple color if protein is present. The disadvantages of the swab test are the determination of the color is based subjective visual inspection and the quantity of protein present is unknown. In this project a quantitative method for reading 3M™ Clean- Trace™ Surface Protein (Allergen) swab tests kits was developed using a micro plate assay. Bovine serum albumin (BSA), egg whites, non-fat dry milk (NFDM), and soy isolate were applied to the swab test at various quantities and the absorbance of the test kit solution was measured at 560nm. With this quantitative approach it was possible to detect 2μg of BSA, 5μg of egg whites, NFDM, and soy isolate. The method provided increased sensitivity from the traditional visual color determination which detected 6ug BSA, 10ug of egg whites, 15ug of soy isolate and inconclusive results for the 15μg of NFDM. The use of this approach will reduce excess cleaning of equipment surfaces for food manufacturers by providing a quantitative result for low quantities of protein, where the visual color determination is subjective or ambiguous

Effect of Cover Crops and Nitrogen Application Timing on Nutrient Loading and Concentration through Subsurface Tile Drainage

It has been estimated that nitrate (NO3-N) leaching from artificially drained agricultural fields in the Upper Mississippi River Basin accounts for approximately 65% of nitrogen (N) delivered annually to the Gulf of Mexico. Therefore, the objectives of this study were to investigate the impact fertilizer application timing and cover crops (CC) adoption on the load and concentration of nutrients in subsurface drainage leachate. This experiment was conducted at the Illinois State University Nitrogen Management Field Station, in Lexington, IL. Treatments include a zero control (no N fertilizer and no cover crop), fall dominated N application (70% fall, 30% spring) with and without CC, and a spring dominated N application (20% fall, 80% spring) with and without CC. A total rate of 224 kg N ha-1 was applied to all treatments, except the zero control. Spring cover crop sampling revealed an average biomass of 1165 kg ha-1 and an average N uptake of 42.5 kg N ha-1. Soil samples were collected in the spring at four separate depths and were analyzed for inorganic N. Regardless of N fertilizer application system, cover crops demonstrated the ability to decrease the concentration of NO3- N 33-62% within the soil profile across the study. The addition of cover crops also reduced the amount of NO3- N within at the lower depths of the soil profile by 17% in the fall system and 37% in the spring system. The Illinois Nutrient Loss Reduction Strategy presents a target reduction in NO3-N losses by 15% by 2025. This data demonstrates that CC have the potential to significantly reduce NO3-N losses in both spring and fall dominated N management systems

AB0378 UPGRADING THERAPY STRATEGY IMPROVES PREGNANCY OUTCOME IN ANTIPHOSPHOLIPID SYNDROME: A COHORT MANAGEMENT STUDY

Background:While it is generally agreed that pregnant APS patients should receive personalized treatment, evidence-based guidelines for these patients continue to be lacking.Objectives:The current study was designed as a management cohort study aiming to evaluate the efficacy and safety of different treatment strategies for pregnant APS patients in the attempt to provide some practical suggestions for attending physicians.Methods:One-hundred-twenty-seven consecutive pregnancies were assessed; 87 (68.5%) with a history of pregnancy morbidity alone were treated with prophylactic low molecular weight heparin (LMWH)+low-dose aspirin (LDA, 100 mg) [Group I] and 40 (31.5%) with a history of thrombosis and/or severe pregnancy complications with therapeutic LMWH+LDA [Group II]. LMWH doses were increased throughout the pregnancies depending on the patients' weight gain, and treatment was switched to a more intensive one at the first sign of maternal/fetal complications. The study's primary outcome was live births.Results:There were no significant differences in live birth rate between Group I (95.4%) and Group II (87.5%). Even, fetal complication rate was similar in the two groups; the Group II nevertheless had a higher prevalence of maternal and neonatal complications (p=0.0005 and p=0.01, respectively) and registered a significantly lower gestational age at delivery and birth weight (p=0.0001 and p=0.0005, respectively). Two patients in Group I switched to Group II therapy, six patients in Group II switched to a more intensive treatment strategy (weekly plasma exchange+ fortnightly intravenous immunoglobulins in addition to therapeutic LMWH+LDA). Comparison of the clinical and laboratory characteristics between patients who had shifted to a more intensive therapy and those who did not showed a significant prevalence of history of thrombosis ± pregnancy morbidity (p=0.02, OR 5.96, 95% CI 1.33-26.62) previous pregnancy complications (p=0.02, OR 8.32, 95% CI 1.67-41.3), triple aPL positivity (p <0.0001, OR 97.13, 95% CI 10.6-890) and pregnancy complications (p<0.0001, OR 197,7, 95% CI 10.57-3699) in upgrading group, instead single aPL positivity significantly prevailed (p=0.003, OR 0.06, 95% CI 0.008-0.58) in non-upgrading group. Logistic regression analysis demonstrated that triple aPL positivity was an independent factor for switching to a more effective therapy protocol (p <0.0001, OR 98, 95% CI 10.7-897.54). All eight switched patients achieved a live birth.Conclusion:Using adjusted LMWH doses and upgrading therapy at the first signs of pregnancy complications led to a high rate of live births in a relatively large group of APS patients. The study outlines the criteria for prescribing appropriate therapy for various subsets of these patients and for switching/upgrading the treatment protocol when it is no longer sufficient. Unfortunately, for the moment there are no evidence-based guidelines on the ideal additional treatment in refractory to conventional therapy APS patients. The present results will hopefully help point the direction of future clinical trials investigating the efficacy and safety of the different therapies on large numbers of APS pregnant patients in order to identify the benefits and limits of different treatment strategies administered from the beginning of pregnancy.Disclosure of Interests:Ariela Hoxha Speakers bureau: Celgene, UCB, Novartis, Sanofi, Werfen, Maria Favaro: None declared, Antonia Calligaro: None declared, Teresa Del Ross: None declared, Alessandra Teresa Ruffatti: None declared, Chiara Infantolino: None declared, Marta Tonello: None declared, Elena Mattia: None declared, Amelia Ruffatti: None declare

Upgrading Therapy Strategy Improves Pregnancy Outcome in Antiphospholipid Syndrome: A Cohort Management Study

The current study evaluates the efficacy and safety of different treatment strategies for pregnant patients with antiphospholipid syndrome. One hundred twenty-seven consecutive pregnancies were assessed; 87 (68.5%) with a history of pregnancy morbidity alone were treated with prophylactic low molecular weight heparin (LMWH) + low-dose aspirin (LDA, 100 mg) (group I) and 40 (31.5%) with a history of thrombosis and/or severe pregnancy complications with therapeutic LMWH + LDA (group II). LMWH doses were increased throughout the pregnancies depending on the patients' weight gain, and treatment was switched to a more intensive one at the first sign of maternal/fetal complications. The study's primary outcome was live births. There were no significant differences in live birth rate between group I (95.4%) and group II (87.5%). Even fetal complication rate was similar in the two groups; group II nevertheless had a higher prevalence of maternal and neonatal complications (p = 0.0005 and p = 0.01, respectively) and registered a significantly lower gestational age at delivery and birth weight (p = 0.0001 and p = 0.0005, respectively). Two patients in group I switched to group II therapy, six patients in group II switched to a more intensive treatment strategy (weekly plasma exchange + fortnightly intravenous immunoglobulins in addition to therapeutic LMWH + LDA). The multivariate analysis uncovered that triple antiphospholipid antibodies positivity was an independent factor leading to a more intensive therapy. All eight switched patients achieved a live birth. Study results revealed that adjusted LMWH doses and switching therapy at first signs of severe pregnancy complications led to a high rate of live births in antiphospholipid syndrome patients

Confirmation of antiphospholipid antibody positivity: a year's results in a cohort of 113 patients

Objective: To evaluate the confirmation rate of antiphospholipid antibodies (aPL), to analyze their behaviour at confirmation time, and to study the clinical value of their confirmation. Methods: Blood samples from 380 subjects, enrolled in this study from June 1, 2007 to May 31, 2008, were tested for anti-cardiolipin (aCL) and anti-beta2glycoprotein (aβ2GPI) antibodies using an ELISA method and for Lupus anticoagulant (LA) using a series of clotting tests. The samples of the 113 subjects resulting positive at the first testing time were assayed again to confirm antiphospholipid positivity. Results: aPL positivity was confirmed in 67 out of the 113 subjects (59.3%). Medium-high antibody levels of all, except IgM aCL, aPL/ELISA had a significantly higher confirmation rate with respect to that in subjects with low levels. The confirmation rate in the category I antibody patients (multiple positivity) was higher than that in the category II antibody subjects (single positivity). LA positivity was confirmed only when it was associated to other aPL. The cut-off of 40 GPL produced a confirmation rate equal to that resulting from a 99th percentile cut-off. Confirmation of aPL positivity made it possible for us to confirm the diagnosis of antiphospholipid syndrome (APS) in 8 out of the 113 subjects originally resulting positive (7,1%). APS clinical features were vascular thrombosis in 4 of these and pregnancy morbidity in the other 4. Conclusions: Our data emphasize aPL positivity confirmation selectivity, and medium-high antibody levels and category I antibodies (multiple positivity) had the best confirmation rates

Clinical significance of fluoroscopic patterns specific for the mitotic spindle in patients with reumatic diseases

Objective: we proposed to determine the clinical significance of anti-NuMA and anti-HsEg5 antibodies in a group of patients affected with rheumatic diseases. Materials and methods: indirect immunofluorescence on HEp-2000 cells at serum dilution of 1:40 was used to examin 26 sera which had previously showed a "mitotic spindle" fluoroscopic pattern type during laboratory routine. Results: 21 sera (80,7%) were identified with NuMA and 5 (19,3%) with HsEg5 patterns alone or associated with other ANA patterns. However only patients with isolated positiveness and that is 15 with NuMA and 4 with HsEg5 stainings were included in this study. Of the NuMA positive patients 5 were affected with arthropathies associated to different forms of thyroiditis, 2 with seronegative arthritis, 2 with antiphospholipid syndrome, 1 with systemic lupus erythematosus (SLE), 1 with rheumatoid arthritis, 1 with sicca syndrome, 1 with undifferentiated connective tissue disease, 1 with Mycoplasma pneumaniae infection and 1 with retinal thrombosis. Of the HsEg5 positive patients 3 were affected with SLE and 1 with seronegative arthritis. Conclusions: NuMA does not prevail in any defined rheumatic disease, while HsEg5 staining were more frequent (75%) in patients affected with SLE all of whom showing high antibody titres

Complement activation in the plasma and placentas of women with different subsets of antiphospholipid syndrome

Problem: As antiphospholipid antibody\u2010positive women with adverse pregnancy outcomes have higher plasma complement activation product levels, and the placentas of women with antiphospholipid syndrome (APS) exhibit C4d complement component deposition, complement activation involvement has been hypothesized in APS pregnancy complications. Method of study: Plasma levels of C5a and C5b\u20109 complement components of 43 APS non\u2010pregnant patients and 17 pregnant APS women were measured using enzyme\u2010 linked immunosorbent assay. The results were compared with those of 16 healthy non\u2010pregnant women and eight healthy pregnant women, respectively. Placenta samples of five APS patients at high risk of pregnancy complications and of five healthy controls were subjected to immunoblotting analysis with specific antibodies to C5b\u20109 and CD46, CD55, CD59 complement regulators. Results: The mean plasma C5a and C5b\u20109 levels were significantly higher in the nonpregnant APS patients with previous thrombosis \ub1 pregnancy morbidity (P = .0001 and P = .0034, respectively) and in the pregnant APS women with adverse outcomes (P = .0093 for both). Similarly, C5b\u20109 amounts were significantly higher in the adverse pregnancy outcome placenta (P = .0115) than in those associated to a favorable outcome. The mean CD46, CD55 and CD59 amounts were, instead, lower, although not always significantly, in the placentas of all the high\u2010risk APS women with respect to the control placentas. Conclusion: Data analysis demonstrated that there was significant complement activation in the more severe subset of APS patients and in only the adverse pregnancy outcome APS women. Further studies will clarify whether the lower CD46, CD55, and CD59 expressions in the APS placentas are limited to only high\u2010risk APS patients

Lupus anticoagulant identifies two distinct groups of patients with different antibody patterns

Background: Whether antibodies directed to β2-Glycoprotein I (aβ2GPI) are responsible for LA activity is not well defined. However, in the absence of such antibodies the molecule responsible for LA phenomenon is unknown. Objective: The aim of this study was the biochemical identification of the target antigen epitope of aPL responsible of LA activity in the absence of aβ2GPI antibodies together with the biological and clinical characteristics of these patients in comparison with classical triple positive patients. Patients/methods: A comparison of patients with LA without (LA+/aβ2GPI−) and those with (LA+/aβ2GPI+) associated aβ2GPI antibodies was performed. Size exclusion chromatography and analytical chromatography were used to identify the molecule with LA activity in patients LA+/aβ2GPI-. Results and conclusions: Analytical size-exclusion chromatography revealed a peak of 996Kd with LA activity perfectly overlapping that of IgM anti phosphatidylserine/prothrombin (aPS/PT) antibodies. Similarly, all the 25 LA+/aβ2GPI− patients were positive for aPS/PT antibodies. LA+/aβ2GPI− compared to 33 LA+/aβ2GPI+ patients turned out to be significantly older, with a lower rate of previous thromboembolic events and a weaker LA activity. Search for aPS/PT and aβ2GPI antibodies in patients with LA is useful to identify two subgroups of LA at different risk of thromboembolic event

THE CROSSTALK BETWEEN THE COMPLEMENT SYSTEM AND THE COAGULATION CASCADE IN THE ANTIPHOSPHOLIPID SYNDROME. PRELIMINARY DATA FROM BASIC RESEARCH.

The association between antiphospholipid antibodies (aPL) and thrombophilic state in antiphospholipid syndrome (APS) is well recognized, but the underlying pathophysiology remains incompletely elucidated. Several findings suggest the role of complement system (CS) in the pathogenesis of antiphospholipid syndrome (APS). The importance of CS in APS is understandable since complement-derived inflammatory mediators increase vascular permeability, activate platelets and promote release of cytokines from monocytesthat favor systemic inflammation and coagulation. It has been demonstrated in a mouse model of aPL-induced pregnancy loss that complement activation can amplify the fetal injury. CS activation has been also documented in patients with APS, but there are far fewer clinical data