A review of the unique features of HDL apoproteins

The human plasma high density lipoproteins (HDL) are a heterogeneous ensemble of five proteins associated with both neutral and polar lipids. The sequences of all five proteins are known. ApoA-I and apoA-II are the major protein components; apoC-I, apoC-II and apoC-III are the minor protein components. All these apoproteins spontaneously recombine with phospholipids to give stable lipid-protein complexes and freely exchange between the two major HDL subclasses, HDL₂ and HDL₃. In addition, ApoC-I, apoC-II, and apoC-III exchange between HDL and very low density lipoproteins. Furthermore, certain HDL apoproteins are activators for plasma enzymes that are important in lipid metabolism. ApoA-I and apoC-I activate lecithin/cholesterol acyltransferase; apoC-II is an activator of lipoprotein lipase. The regions of apoC-I and apoC-II that are involved in the activation of these enzymes have been localized with synthetic peptides. Studies of synthetic and native fragments of apoA-II, apoC-I, apoC-II, and apoC-III as well as model lipid-binding peptides have identified specific regions with structural features common to lipid-binding proteins. These special properties, which include helical potential, sequences with a critical amphipathic length, and high hydrophobicity of the nonpolar side of the amphipathic helix, are the determinants of HDL structure and metabolism.     

Isosteric heat of sorption of CO2 in poly(ethylene terephthalate)

Sorption isotherms for carbon dioxide in poly(ethylene terephthalate) have been measured at 35–55°C. The isotherms were measured gravimetrically on a Mettler Thermoanalyzer-1 from vacuum to 1 atmosphere. The sorption data were used to generate sorption isotherms from which the isosteric heat of sorption of CO₂ in PET was determined. At 45°C the isosteric heat of sorption increases from −10 kcal/mole at a concentration of 0.5 cm³ (STP)/cm³ (polymer) to −8 kcal mole⁻¹ at a concentration of 1.5 cm³ (STP)/cm³ (polymer). It has been reported in the literature that the isosteric heat of sorption for this system decreased through a minimum before increasing with increasing concentration. Our measurement of the low-pressure sorption isotherms shows that this is not the case.     

A Hidden‐picture Puzzles Generator

A hidden‐picture puzzle contains objects hidden in a background image, in such a way that each object fits closely into a local region of the background. Our system converts image of the background and objects into line drawing, and then finds places in which to hide transformed versions of the objects using rotation‐invariant shape context matching. During the hiding process, each object is subjected to a slight deformation to enhance its similarity to the background. The results were assessed by a panel of puzzle‐solvers.     

Digital Intuition: Applying Common Sense Using Dimensionality Reduction

Understanding the world we live in requires access to a large amount of background knowledge: the commonsense knowledge that most people have and most computer systems don't. Many of the limitations of artificial intelligence today relate to the problem of acquiring and understanding common sense. The Open Mind Common Sense project began to collect common sense from volunteers on the Internet starting in 2000. The collected information is converted to a semantic network called ConceptNet. Reducing the dimensionality of ConceptNet's graph structure gives a matrix representation called AnalogySpace, which reveals large-scale patterns in the data, smoothes over noise, and predicts new knowledge. Extending this work, we have created a method that uses singular value decomposition to aid in the integration of systems or representations. This technique, called blending, can be harnessed to find and exploit correlations between different resources, enabling commonsense reasoning over a broader domain.     

Purification, characterization, and high performance liquid chromatography assay of Salmonella glucose-1-phosphate cytidylyltransferase from the cloned rfbF gene

We report the purification and characterization of glucose-1-phosphate cytidylyltransferase, the first of five enzymes committed to biosynthesis of CDP-D-abequose from Salmonella enterica strain LT2. The purification was greatly facilitated by using a cloned rfbF gene encoding this enzyme. Pure enzyme was obtained by 64-fold enrichment in three chromatography steps. The NH2-terminal sequence of the purified enzyme was in agreement with the sequence predicted from the nucleotide sequence of the rfbF gene. The SDS-polyacrylamide gel electrophoresis estimated subunit M(r) of 31,000 agrees well with the M(r) of 29,035 calculated from the amino acid composition deduced from the nucleotide sequence of the rfbF gene. The glucose-1-phosphate cytidylyltransferase catalyzes a reversible bimolecular group transfer reaction and steady-state kinetic measurements, including product inhibition patterns, indicate that this reaction proceeds by a "ping-pong" type of mechanism. The Km values for CTP, alpha-D-glucose 1-phosphate, CDP-D-glucose, and pyrophosphate are 0.28, 0.64, 0.11, and 1.89 mM, respectively.     

Ex vivo staining of metastatic lymph nodes by class I major histocompatibility complex tetramers reveals high numbers of antigen-experienced tumor-specific cytolytic T lymphocytes

Characterization of cytolytic T lymphocyte (CTL) responses to tumor antigens has been impeded by a lack of direct assays of CTL activity. We have synthesized reagents ("tetramers") that specifically stain CTLs recognizing melanoma antigens. Tetramer staining of tumor-infiltrated lymph nodes ex vivo revealed high frequencies of tumor-specific CTLs which were antigen-experienced by surface phenotype. In vitro culture of lymph node cells with cytokines resulted in very large expansions of tumor-specific CTLs that were dependent on the presence of tumor cells in the lymph nodes. Tetramer-guided sorting by flow cytometer allowed isolation of melanoma-specific CTLs and confirmation of their specificity and their ability to lyse autologous tumor cells. Our results demonstrate the value of these novel reagents for monitoring tumor-specific CTL responses and for generating CTLs for adoptive immunotherapy. These data also indicate that strong CTL responses to melanoma often occur in vivo, and that the reactive CTLs have substantial proliferative and tumoricidal potential.     

Determination of fat in olestra-containing savory snack products by capillary gas chromatography

A quantitative method to determine fat in olestra-containing savory snack products was validated within the AOAC Peer-Verified Methods Program. The method may be used to demonstrate compliance with the guidelines of the U.S. Nutrition Labeling and Education Act for labeling products as "fat free" or "low fat." The method can measure total and saturated fat in savory snacks when present at levels of 0.2-10 g total fat and 0.1-3 g saturated fat per 30 g serving. The method is standardized to measure C6-C24 fatty acids. Extraction of olestra-containing savory snack samples with chloroform-methanol (modified AOAC Official Method 983.23) yields a lipid extract containing the total fat and olestra. The extracted lipid is hydrolyzed by lipase, yielding fatty acids and unreacted olestra. The fatty acids are precipitated as calcium soaps. Olestra is extracted from insoluble soaps with hexane and then discarded. The isolated soaps are converted back into fatty acids with hydrochloric acid and extracted with hexane. The isolated fatty acids are converted to methyl esters with boron trifluoride-methanol and quantitated by capillary gas chromatography using internal standard. Test samples were prepared by blending olestra-containing and full-fat (triglyceride) snacks to obtain 6 levels of spiking (0-10 g total fat added/30 g serving) in potato chips, potato crisps, cheese puffs, and nacho cheese-flavored corn chips. Results were linear (r2 > 0.997) between 0 and 10 g fat/30 g serving for each product matrix. Mean recovery was 101 +/- 6% standard deviation (SD) for total fat and 104 +/- 6% SD for saturated fat. Mean recovery by peer laboratory was 88 +/- 5% SD for total fat and 95 +/- 4% SD for saturated fat in potato chips (0-3 g total fat added/30 g serving). Two sets of 10 replicates of potato chips (0.5 g total fat/30 g serving and 0.16 g saturated fat/30 g serving) and potato crisps (0.5 g total fat/30 g serving and 0.16 g saturated fat/30 g serving) were analyzed by submitting and peer laboratories. Repeatability relative standard deviations ranged from 3.90 to 7.33% for total fat and from 4.01 to 11.53% for saturated fat. Reproducibility relative standard deviations were 7.33% (total fat, potato chips), 7.15% (total fat, potato crisps), 11.36% (saturated fat, potato chips), and 13.50% (saturated fat, potato crisps).     

A clinical evaluation in children of the Pharmacia ImmunoCAP system for inhalant allergens

BACKGROUND: The Pharmacia ImmunoCAP system (CAP) for assaying serum IgE specific antibodies was evaluated in a clinical setting against skin-prick test (SPT) performed using Dome/Hollister-Steir allergen extracts. The five common inhalant allergens D. pteronyssinus, D. farinae, mould mix, grass mix and cat epithelium were tested concurrently by both methods in 167 children aged 7.5-12 years. The specific SPT for D. pteronyssinus and D. farinae were also tested against the CAP house dust mite (HDM) mix. OBJECTIVE: The purpose of the study was to determine the sensitivity and specificity of the Pharmacia ImmunoCAP system for detecting serum IgE specific antibodies to inhalant allergens in a clinical setting, using SPT result as the "gold standard'. METHODS: The SPTs were performed using Dome/Hollister-Steir allergen extracts. The serum IgE specific antibodies were quantitated using the radioimmunoassay version of the Pharmacia ImmunoCAP system. A history of allergic disease was assessed using a validated questionnaire. RESULTS: SPT gave more positive reactions than CAP with the exception of cat epithelium. The concordance between SPT and CAP results was 91% over all the tests. The concordance with SPT was slightly higher for the specific CAP for D. pteronyssinus and D. farinae (93% and 95% respectively) than for the CAP HDM mix (86% and 90% respectively). There was a higher proportion of positive results for both SPT and CAP in the 115 children defined as having a history of allergic disease. Using SPT defined allergy as the gold standard, the sensitivity of the CAP system was 87% for the two specific house dust mites but was lower for cat epithelium (67%), mould mix (59%) and grass mix (46%). The sensitivity of the CAP system improved for D. pteronyssinus (96%) and the HDM mix (91%) when tested in subjects defined as having a history of allergy associated disease. The specificity of the CAP system showed less variation between allergens and ranged from 90-99%. CONCLUSION: The results of this study of children aged 7.5-12 years demonstrate that, for the inhalant allergens tested, the Pharmacia ImmunoCAP system performs well in the setting of known allergic disease.