SUMMARY. Against a background of growing interest in more sensitive assays for quantifying various acute phase proteins, we evaluated the performance of recently developed tests for C-reactive protein (CRP), serum amyloid A (SAA) and mannosebinding protein (MBP) on the Behring nephelometer II (BN II). Sample results outside the calibration ranges of 3·5 to 220mgjL for CRP, 3·3 to 215mgjL for SAA and 0·09 to 5'6mgjL for MBP were automatically re-measured at another dilution. The lower limits of detection were 0·01, 0·7 and 0·01 mgjL for CRP, SAA and MBP, respectively. The coefficients of variation (CY) for intra-(n~20) and inter-(n~15) assay precision were < 5·2% and < 8,5%, respectively, for the three proteins at concentrations representing low, normal and high. Linearity for each method was within 5% of the expected values throughout the calibration range. We observed no significant interference from bilirubin (up to 300 mgjL) or haemoglobin (up to 10 gj L) for the three tests. Method comparison studies performed for CRP and SAA yielded the following results: y (CRP on BN II) = 0'75x (ELISA, Hemagen) -0'25mgjL (1'=0'981, Sy/x=2·1 mgjL; J' (SAA on BNII)= 1·44x (ELISA, Hemagen) -9·9 mgjL (r = 0·972, Sy/x = 6·9 mg/L), where ELISA is enzyme-linked immunosorbent assay. Reference intervals established in 261 adult blood donors (aged 36·2±9·0 years) were found to be log-normal with 2'5th, 50th and 97·5th centilcs of <0,17,1·00 and 10·1 mgjL for CRP, <0·84,2,10 and 9'70mgjL for SAA; and 0,30, 1·28 and 4·10mgjL for MBP. We observed no relationship with CRP concentration and age; however, SAA levels increased with age while MBP levels decreased. The BN 11 provides a simple, rapid and sensitive system for measuring CRP, SAA and MBP in human serum.

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Experiments reported here indicate a crude soybean extract, if defatted with acetone, effectively blocks cell transformation in vitro. An active component of this crude extract is the Bowman-Birk trypsin and chymotrypsin inhibitor. The chymotrypsin-inhibitory region of the Bowman-Birk inhibitor is responsible for suppressing in vitro transformation. Another low molecular weight soybean trypsin inhibitor does' not significantly suppress transformation. The Bowman-Birk inhibitor (i) has an irreversible effect on the transformation process, (ii) can suppress radiation-induced transformation even when added to cultures many days after the carcinogen exposure, and (iii) is effective in its ability to suppress transformation when present in the medium at a concentration as low as 0.125 nM.We have reported that protease inhibitors, including the Bowman-Birk trypsin and chymotrypsin inhibitor from soybeans, have the ability to suppress x-ray-induced malignant transformation in C3H/10T½ cells (refs. 1-4; reviewed (4,9). We now report that (i) a crude extract of the inhibitor, which can be obtained in reasonable quantities for use in animal carcinogenesis experiments, has the ability to inhibit transformation in vitro; (ii) other soybean protease inhibitors, or other compounds present in our crude extract, lack the ability to suppress transformation in vitro; (iii) the chymotrypsin-inhibitory site of BBI is involved in the suppressive effects of the BBI on transformation; (iv) the lowest effective dose of the protease inhibitor preparations that has the ability to suppress transformation in vitro is 0.125 nM. MATERIALS AND METHODSThe C3H/10T½2 transformation assay was developed by Reznikoff et al. (10,11) and modified for use in radiationinduced transformation experiments (1-5). The details of our experimental techniques for radiation-induced transformation experiments using C3H/10T½/ cells and protease inhibitors have been described (1)(2)(3)(4)(5). Stock cultures were maintained in 60-mm Petri dishes and were passed by subculturing at a 1:20 dilution every 7 days. The cells used were in passages 7-12. They were grown in a humidified 5% CO2 in air atmosphere at 37°C in Eagle's basal medium supplemented with 10% heat-inactivated fetal calf serum and gentamycin. In all transformation experiments, the concentration of serum was reduced to 5% on day 10 and was maintained at this concentration throughout the remainder of the 6-week assay period. Plating efficiencies (PEs) were determined from three plates seeded with a cell density one-fifth of that of the plates used for the transformation assay; these cultures were terminated at 10 days. The various treatment toxicities were considered in the design of the experiments so that all of the dishes used for the transformation assay contained "300 viable cells per dish (initially). Types 2 and 3 foci were scored as transformants; type 3 cells have been found to be tumorigenic in 80-100% of inoculated mice; type 2 cells are tumorigenic in 60-75% of inoculated mice (1-5)...

BackgroundEnsuring the quality of malaria medicines is crucial in working toward malaria control and eventual elimination. Unlike other validated tests that can assess all critical quality attributes, which is the standard for determining the quality of medicines, basic tests are significantly less expensive, faster, and require less skilled labour; yet, these tests provide reproducible data and information on several critical quality attributes, such as identity, purity, content, and disintegration. Visual and physical inspection also provides valuable information about the manufacturing and the labelling of medicines, and in many cases this inspection is sufficient to detect counterfeit medicines. The Promoting the Quality of Medicines (PQM) programme has provided technical assistance to Amazon Malaria Initiative (AMI) countries to implement the use of basic tests as a key screening mechanism to assess the quality of malaria medicines available to patients in decentralized regions.MethodsTrained personnel from the National Malaria Control Programmes (NMCPs), often in collaboration with country’s Official Medicine Control Laboratory (OMCL), developed country- specific protocols that encompassed sampling methods, sample analysis, and data reporting. Sampling sites were selected based on malaria burden, accessibility, and geographical location. Convenience sampling was performed and countries were recommended to store the sampled medicines under conditions that did not compromise their quality. Basic analytical tests, such as disintegration and thin layer chromatography (TLC), were performed utilizing a portable mini-laboratory.ResultsResults were originally presented at regional meetings in a non-standardized format that lacked relevant medicines information. However, since 2008 information has been submitted utilizing a template specifically developed by PQM for that purpose. From 2005 to 2010, the quality of 1,663 malaria medicines from seven AMI countries was evaluated, mostly collected from the public sector, 1,445/1,663 (86.9%). Results indicate that 193/1,663 (11.6%) were found not to meet quality specifications. Most failures were reported during visual and physical inspection, 142/1663 (8.5%), and most of these were due to expired medicines, 118/142 (83.1%). Samples failing TLC accounted for 27/1,663 (1.6%) and those failing disintegration accounted for 24/1,663 (1.4%). Medicines quality failures decreased significantly during the last two years.ConclusionsBasic tests revealed that the quality of medicines in the public sector improved over the years, since the implementation of this type of quality monitoring programme in 2005. However, the lack of consistent confirmatory tests in the quality control (QC) laboratory, utilizing methods that can also evaluate additional quality attributes, could still mask quality issues. In the future, AMI countries should improve coordination with their health authorities and their QC lab consistently, to provide a more complete picture of malaria medicines quality and s...

Recent efforts to improve the serologic diagnosis of Lyme disease have included the use of a synthetic peptide (C6) that reproduces the sequence of invariable region 6 of VlsE, the variable surface antigen of Borrelia burgdorferi. In the present study, the diagnostic performance of DiaSorin's recombinant VlsE-based chemiluminescence immunoassay in 1,947 human serum samples was evaluated. Sensitivity was determined using two serum panels from the CDC. For panel I, we observed sensitivities of 68.4% and 75.6% for subjects with early, localized (n ‫؍‬ 19) or disseminated (n ‫؍‬ 41) disease, respectively. For panel II, we observed sensitivities of 61.5% and 100% for subjects with early (n ‫؍‬ 26) or late-stage (n ‫؍‬ 11) disease, respectively. We observed a specificity of 99.5% for healthy donors (n ‫؍‬ 600) living either in regions of the United States where the disease is endemic or in regions where it is not endemic. Overall, specificity among 207 potentially cross-reactive sera from subjects who had other spirochetal infections, nonspirochetal infections including bacterial and viral infections, or autoimmune or neurologic disease; who were positive for rheumatoid factor or anti-mouse antibodies; or who had been previously vaccinated for Lyme disease was 93.7%. In a direct comparison of 1,038 prospectively collected samples for Lyme disease testing we observed a relative sensitivity of 70%, a relative specificity of 99.1%, and an overall agreement of 97.1% between the DiaSorin recombinant VlsE chemiluminescence immunoassay and the Immunetics peptide-based C6 enzyme-linked immunosorbent assay.