Perceived 'healthiness' of foods can influence consumers' estimations of energy density and appropriate portion size.

OBJECTIVE: To compare portion size (PS) estimates, perceived energy density (ED) and anticipated consumption guilt (ACG) for healthier vs standard foods. METHODS: Three pairs of isoenergy dense (kJ per 100 g) foods-healthier vs standard cereals, drinks and coleslaws-were selected. For each food, subjects served an appropriate PS for themselves and estimated its ED. Subjects also rated their ACG about eating the food on a scale of 1 (not at all guilty) to 5 (very guilty). RESULTS: Subjects (n=186) estimated larger portions of the healthier coleslaw than that of the standard version, and perceived all healthier foods to be lower in ED than their standard alternatives, despite being isoenergy dense. Higher ACG was associated with the standard foods. Portion estimates were generally larger than recommendations and the ED of the foods was underestimated. CONCLUSIONS: The larger portions selected for the 'reduced fat' food in association with lower perceived ED and ACG suggests that such nutrition claims could be promoting inappropriate PS selection and consumption behaviour. Consumer education on appropriate portions is warranted to correct such misconceptions.

Supermarket own brand foods: lower in energy cost but similar in nutritional quality to their market brand alternatives.

BACKGROUND: The present study aimed to compare the nutritional quality (NQ) and energy costs (EC) (£ MJ(-1) ) of own brand (OB) versus market brand (MB) foods in 2010 and 2012. METHODS: A list of processed foods (n = 32) was identified based on the most frequently consumed foods in the UK. Total fat, saturated fat, sugars, salt and energy density (ED) (kJ g(-1) ) in 2010 and 2012 were compared for six OB and one MB version of each food using a NQ scoring method based on the Food Standards Agency's Traffic Light System (TLS). Additional information (fruit, vegetable and nut content; protein; fibre and sodium) was recorded in 2012, and NQ was assessed using the Food Standards Agency's nutrient profiling model (NPM). The EC of the food baskets (FB) was compared in 2010 and 2012. RESULTS: There were no differences in overall NQ between OB and MB FB in 2010 (TLS, P = 0.978) or 2012 (TLS, P = 0.840; NPM, P = 0.696). However, the MB FB was highest in EC in 2010 and 2012 (both P < 0.001). There was an inverse relationship between the ED and EC of the MB foods in 2010 (r = -0.484; P = 0.005) and 2012 (r = -0.452; P = 0.009). CONCLUSIONS: The MB FB was higher in EC than the OB FB in 2010 and 2012 but not superior in overall NQ based on both the TLS and NPM.

The type, amount, location, and energy transfer properties of the carotenoid in reaction centers from Rhodopseudomonas sphaeroides.

Analysis of photosynthetic reaction centers from Rhodopseudomonas sphaeroides strains 2.4.1 and Ga shows that each contains approx. 1 mol of a specific carotenoid per mol of reaction center. In strain 2.4.1. the carotenoid is spheroidene (1-methoxy-3,4-didehydro-1,2,7',8',-tetrahydro-psi,psi-carotene); in strain Ga, it is chloroxanthin (1-hydroxy-1, 2, 7', 8'-tetrahydro-psi,psi-carotene). The carotenoid is bound to the same pair of proteins as are the bacteriochlorophylls and bacteriopheophytins of the reaction center. This binding induces strong circular dichroism in the absorption bands of the carotenoid. The carotenoid is close enough to the other pigments of the reaction center so that light energy transfers efficiently from the carotenoid to the bacteriochlorophyll, sensitizing bacteriochlorophyll fluorescence. The fluorescence polarization spectrum of the reaction centers shows that the transition vectors for the visible absorption bands of the carotenoid lie approximately parallel to the 600 nm (Qx) transition of the bacteriochlorophyll complex.

The Fab and Fc fragments of IgA1 exhibit a different arrangement from that in IgG: a study by X-ray and neutron solution scattering and homology modelling.

Human immunoglobulin A (IgA) is an abundant antibody that mediates immune protection at mucosal surfaces as well as in plasma. The IgA1 isotype contains two four-domain Fab fragments and a four-domain Fc fragment analogous to that in immunoglobulin G (IgG), linked by a glycosylated hinge region made up of 23 amino acid residues from each of the heavy chains. IgA1 also has two 18 residue tailpieces at the C terminus of each heavy chain in the Fc fragment. X-ray scattering using H2O buffers and neutron scattering using 100 % 2H2O buffers were performed on monomeric IgA1 and a recombinant IgA1 that lacks the tailpiece (PTerm455). The radii of gyration RG from Guinier analyses were similar at 6.11-6.20 nm for IgA1 and 5.84-6.16 nm for PTerm455, and their cross-sectional radii of gyration RXS were also similar. The similarity of the RG and RXS values suggests that the tailpiece of IgA1 is not extended outwards in solution. The IgA1 RG values are higher than those for IgG, and the distance distribution function P(r) showed two distinct peaks, whereas a single peak was observed for IgG. Both results show that the hinge of IgA1 results in an extended Fab and Fc arrangement that is different from that in IgG. Automated curve-fit searches constrained by homology models for the Fab and Fc fragments were used to model the experimental IgA1 scattering curves. A translational search to optimise the relative arrangement of the Fab and Fc fragments held in a fixed orientation resembling that in IgG was not successful in fitting the scattering data. A new molecular dynamics curve-fit search method generated IgA1 hinge structures to which the Fab and Fc fragments could be connected in any orientation. A search based on these identified a limited family of IgA1 structures that gave good curve fits to the experimental data. These contained extended hinges of length about 7 nm that positioned the Fab-to-Fab centre-to-centre separation 17 nm apart while keeping the corresponding Fab-to-Fc separation at 9 nm. The resulting extended T-shaped IgA1 structures are distinct from IgG structures previously determined by scattering and crystallography which have Fab-to-Fab and Fab-to-Fc centre-to-centre separations of 7-9 nm and 6-8 nm, respectively. It was concluded that the IgA1 hinge is structurally distinct from that in IgG, and this results in a markedly different antibody structure that may account for a unique immune role of monomeric IgA1 in plasma and mucosa.

Unaggregated serum IgA binds to neutrophil Fc alpha R at physiological concentrations and is endocytosed but cross-linking is necessary to elicit a respiratory burst.

My43 is a monoclonal antibody directed against Fc alpha Rs on monocytes that also recognizes neutrophil (PMN) Fc alpha Rs and is able to elicit a respiratory burst in purified cells. It appears to be directed against the immunoglobulin A (IgA) binding site of Fc alpha Rs or an epitope in close proximity, since IgA and My43 compete for binding to PMNs. My43 immunoblotted Fc alpha R that had been affinity purified from PMN membranes and immunoprecipitated a 50-70 kDa protein from radiolabeled PMN membranes, apparently identical to that purified on IgA-Sepharose. These results suggest the presence of a single class of Fc alpha R on PMNs. Binding of unaggregated monomeric or dimeric serum IgA to Fc alpha Rs on PMNs occurs at physiological concentrations, suggesting that in vivo Fc alpha Rs are saturated with ligand. This binding does not elicit a respiratory burst. Purified PMNs do not express surface IgA, since receptor-bound IgA is lost from the cell surface by endocytosis at room temperature although not at 4 degree C. Rapid endocytosis of receptor-bound IgA has been demonstrated by flow cytometry and confocal microscopy. Unoccupied receptor that is able to bind IgA or My43 is subsequently reexpressed. Cross-linking of surface-bound IgA using F(ab')2 anti-alpha chain antibodies triggers an oxidative burst. After internalization of the surface IgA the same F(ab')2 anti-alpha chain antibodies do not trigger the burst.

CD66-dependent neutrophil activation: a possible mechanism for vascular selectin-mediated regulation of neutrophil adhesion.

We have examined the role of CD66 in the modulation of neutrophil adhesion and effector function. Engagement of neutrophil CD66 with anti-carcinoembryonic antigen (anti-CEA) Ig results in activation-associated phenomena including shape change, activation of beta 2-integrins, and priming of the respiratory burst. Anti-CEA Ig-treated neutrophils underwent transient shape change distinct from that induced by formyl-Met-Leu-Phe (fMLP). fMLP stimulated beta 2-integrin up-regulation and 70% loss of L-selectin, whereas only low-level up-regulation of the beta 2-integrins, without loss of L-selectin, occurred with anti-CEA Ig. Anti-CEA F(ab')2 fragments and whole Ig augmented beta 2-integrin-dependent adhesion. Anti-CEA Ig-induced beta 2-integrin activation was transient, whereas fMLP-induced activation persisted longer. Although they did not cause a significant increase in respiratory burst activity, CEA Ig and F(ab')2 fragments of antibody primed neutrophils so that the subsequent fMLP-induced respiratory burst was significantly increased.

The activation of C3 and C4 in human serum by immune complexes containing mouse monoclonal antibodies of different isotype and affinity: effects on solubilisation.

Radioimmunoassays for C3a and C4a have been used to measure the activation of complement in human serum by immune complexes containing DNP-BSA and each of 11 mouse anti-DNP monoclonal antibodies of varied isotype and affinity. When preformed complexes were added to serum, those containing IgG2 or IgM were potent activators of C4, whilst IgG1 complexes were less efficient. C3 activation in normal serum was similar for complexes containing IgG1, IgG2a, IgG2b or IgM. IgA complexes did not activate C3 or C4. Solubilisation of complexes was greatest for IgM and IgG2b and least for IgG2a and IgA. In serum containing Mg2+ EGTA C4 activation was abolished and the amount of C3 activation was lower for all IgG and IgM complexes. Antibodies of the same isotype did not necessarily activate complement to the same extent. Unexpectedly, three of the four IgMs activated C3 in EGTA. For IgMs, neither complement activation nor solubilisation correlated with affinity. For IgG1 antibodies, solubilisation was inversely proportional to affinity. C3 or C4 activation did not correlate with affinity.

The effect of antibody isotype on the activation of C3 and C4 by immune complexes formed in the presence of serum: correlation with the prevention of immune precipitation.

Radioimmunoassays for C3a and C4a have been used to measure the activation of complement during the formation of immune complexes in human serum by the interaction of DNP-BSA and each of 11 mouse anti-DNP monoclonal antibodies of varied isotype and affinity. Those containing IgG2 or IgM were potent activators of C4, whilst IgG1 containing complexes were less efficient. C3 activation in normal serum was similar for complexes containing IgG1, IgG2a, IgG2b or IgM. IgA complexes did not activate C3 or C4. All complexes except those containing IgA precipitated more slowly in serum than in buffer. IgG2 antibodies were potent activators despite being very slow to precipitate in buffer. In serum containing EGTA activation of C4 was abolished and precipitation of complexes occurred at the same rate as in buffer. Nevertheless, C3 activation still occurred by the alternative pathway for all IgG and IgM complexes. Antibodies of the same isotype did not necessarily activate complement to the same extent. The ranking of the ability to activate complement was the same as that observed when performed complexes containing the same antibodies were added to serum. The levels of C4a generated were similar under both conditions but for most antibodies more C3a was generated by preformed complexes.

Dissociation of primary antigen-antibody bonds is essential for complement mediated solubilization of immune precipitates.

The role of dissociation of primary antigen-antibody bonds in the solubilization of immune complexes (IC) has been investigated using photo-affinity crosslinked IC comprising NAP15-BSA and murine monoclonal anti-DNP antibodies. Non-covalently linked IC were solubilized rapidly when incubated with normal human serum (NHS), whereas covalently-linked IC were solubilized poorly or not at all. The rate and extent of complement activation produced by incubating covalently-linked and non-covalently linked IC with NHS was similar as assessed by the production of the C1s:C1-inhibitor, C3:properdin and C5b-9 complexes and the anaphylatoxins C4a and C3a. Thus, the inability of serum to solubilize photo-affinity crosslinked IC must be due to failure of dissociation of primary antigen-antibody bonds.

The use of spent renal dialysis membranes for the isolation of large numbers of human neutrophils for biochemical studies. Application to purification of the myeloid IgA receptor (Fc alpha R).

Human neutrophils (PMN) can be eluted from spent Cuprophan renal dialysis membranes in large numbers (10(9)-10(10) per dialyser cartridge) and in relatively high purity by washing the membranes with 0.35 M NaCl. This offers the possibility of isolating relatively large amounts (10(-4)-10(-3) g) of minor PMN proteins such as those expressed on the cell surface. Here the technique is applied to the purification of the neutrophil IgA receptor (Fc alpha R). Affinity chromatography on IgA-Sepharose of NP-40 extracts of 125I-labelled PMN isolated from fresh venous blood routinely gave a receptor preparation showing one diffuse band, Mr 50-70 kDa, upon analysis by SDS-PAGE and autoradiography. When the same method was used with larger numbers of unlabelled PMN from fresh venous blood or renal dialysis membranes a preparation was obtained which gave multiple bands upon analysis by SDS-PAGE silver stained gels due to contamination of the receptor with cytoplasmic proteins which bound non-specifically to the IgA-Sepharose. Most of these contaminants could be removed by chromatography of the IgA-Sepharose eluates on wheat germ agglutinin-Sepharose.

Purification and characterization of human immunoglobulin IgA1 and IgA2 isotypes from serum.

A method is described for the simultaneous purification of IgA1 and IgA2 from human serum. Ammonium sulphate precipitation, gel filtration and ion-exchange chromatography on DEAE-Sephacel yielded a partially purified IgA preparation which was separated quantitatively into IgA1 and IgA2 by affinity chromatography on jacalin-Sepharose. The IgA1 which bound to the jacalin was eluted with 0.8 M D-galactose. The IgA1 preparation was apparently homogeneous by SDS-PAGE but contained a trace of C1-inhibitor and a second protein detected by immunoelectrophoresis. The IgA2 which did not bind to the jacalin was purified to apparent homogeneity by chromatography on columns of Protein G-Sepharose, Fastflow-S Sepharose and Superose 6. Typical yields were 95% and 58% for IgA1 and IgA2 respectively or 253 mg and 24 mg per 100 ml serum. The IgA1 and IgA2 were characterised by their reactivity with isotype specific monoclonal antibodies and sensitivity to bacterial proteinases. The IgA2 preparation apparently contained both allotypes, IgA2m(1) and IgA2m(2).

Psychosocial functioning of mothers of malnourished children.

The relationship between infant malnutrition and maternal psychosocial behavior was explored by comparing mothers of malnourished children with mother whose children were matched for age and family income but were not malnourished. The mothers were interviewed and asked to describe their relationships with their children, their children's fathers, extended families, friends, and employers. The mothers of malnourished children described more chronically disrupted lives. Their housing conditions and employment records reflected disorganization. They had fewer social contacts except with extended families who supervised excessively. The fathers of their babies were either not present or unsupportive. Relationships were more stereotyped, transient, and focused on material aspects. The mothers narcissistic concerns took precedence over the needs of their children. Nearly all the mothers, including the controls, had suffered severe deprivation in childhood. Some mothers of malnourished children were apathetic and dependent, whereas others were manipulative and evasive. These findings were consistent with those previously described for mothers of children with "failure-to-thrive" in affluent countries.

Neutrophil NADPH oxidase does not assemble on macropinocytic vacuole membranes.

NADPH oxidase assembly is considered to occur mainly on the plasma membrane and phagolysosome membranes and to a lesser extent in intracellular granules. Stimulation of neutrophils with phorbol 12-myristate 13-acetate (PMA) in conjunction with chemiluminescent substrates has been widely used as a model to understand the sub-cellular locations of NADPH-oxidase activity. Interpretation of data from such studies is complicated by observations that PMA does not trigger phagocytosis but results in formation of large macropinocytic vacuoles. Here we show by laser-scanning confocal microscopy that PMA triggers uptake of lucigenin into macropinocytic vacuoles and that no chemiluminescence (CL) can be detected at this location when NADPH oxidase assembly on the plasma membrane still occurs. This shows that the macropinocytic vacuole membrane is distinct from the phagocytic vacuole membrane although both compartments are derived from the plasma membrane.

Highly selective Diels-Alder reactions of dienophiles with 1, 3-cyclohexadiene mediated by Yb(OTf)(3).2H(2)O and ultrahigh pressures.

[reaction: see text] Ultrahigh pressures and catalytic Yb(OTf)(3).2H(2)O were found to mediate Diels-Alder reactions of various electron-deficient dienophiles with 1,3-cyclohexadiene to produce endo-bicyclo[2.2. 2]oct-2-enes in moderate to excellent yield and selectivity. The proposed total synthesis of hapalindole Q based on bicyclo[2.2. 2]oct-2-ene construction by Diels-Alder reaction and subsequent olefin cleavage is outlined. Preliminary results demonstrating the viability of this strategy are presented.

Total synthesis of (+/-)-hapalindole Q.

[reaction: see text] The total synthesis of the antibacterial and antimycotic alkaloid hapalindole Q has been achieved in eight steps and 12.4% overall yield. The key step involves a regio- and diastereoselective Diels-Alder reaction to afford a bicyclo[2.2.2]oct-2-ene. This cycloadduct was subsequently dihydroxylated, cleaved, and converted to the natural product.

The Diels-Alder reactions of quinone imine ketals: a versatile synthesis of highly substituted 5-methoxyindoles.

[reaction: see text]. N-benzoylated quinone imine ketals undergo smooth cycloadditions in a [4 + 2] sense to yield the expected cycloadducts. The crude cycloadducts, when subjected to a short series of simple transformations, produce synthetically useful quantities of 5-methoxyindoles in excellent overall yields.

Expression in normal adult, fetal, and neoplastic tissues of a carbohydrate differentiation antigen recognised by antigranulocyte mouse monoclonal antibodies.

The distribution in paraffin fixed human tissues of a carbohydrate antigen defined by two monoclonal antibodies raised against human granulocytes has been studied by means of an immunoperoxidase technique. In addition to granulocytes, the antigen has been detected in adult tissues on identifiable cell types of the stomach, kidney, adrenal medulla, and brain and on the mucins of the gastrointestinal tract and other secretions. In fetal tissue, epithelial cells of the alimentary tract, lung, brain, and kidney express the antigen. Adenocarcinoma of the colon, stomach, breast, and lung are stained strongly, as are other types of lung cancer. The monoclonal antibodies give a staining pattern similar but not identical to other monoclonal antibodies raised against granulocytes or neoplastic cell lines which recognise the antigen 3-fucosyl N-acetyllactosamine. The use of antibodies against this oncofetal antigen in the study of differentiation and as tumour markers is discussed.

Expression of CD15 antigen in urinary bladder transitional cell carcinoma.

The biopsy specimens of 91 patients between the ages of 38 and 94 with transitional cell carcinoma of the bladder were retrospectively reviewed to determine if the expression of CD15 antigen detected by a monoclonal antibody MC2 was correlated with prognosis. Expression was variable, ranging from strong expression of the antigen by only the superficial cells in well differentiated papillary lesions to weak expression by most cells in solid or invasive tumours. In the invasive component there was a correlation between MC2 expression and tumour type, suggesting that the cell surface carbohydrate detected by MC2 may have a role in cell adhesion. There was no correlation between staining and survival. It is concluded that tumour type, grade, and stage remain the best prognostic indicators of urothelial tumours.

Fathers and child neglect.

OBJECTIVE: To examine the association between father involvement and child neglect. DESIGN: Cohort study. SETTING: Participants were recruited from an inner-city pediatric primary care clinic and a clinic for children at risk for human immunodeficiency virus infection in a teaching hospital. PARTICIPANTS: Mothers and fathers or father figures, and 244 five-year olds participating in a longitudinal study. MAIN OUTCOME MEASURES: Child neglect measured via home observation, a videotaped mother-child interaction, and child protective services reports. RESULTS: A father or father figure was identified for 72% of the children. Rates of neglect ranged between 11% and 30%. Father absence alone was not associated with neglect. However, in families with an identified and interviewed father, a longer duration of involvement (P<.01), a greater sense of parenting efficacy (P<.01), more involvement with household tasks (P<.05), and less involvement with child care (P<.05) were associated with less neglect. The overall model explained 26.5% of the variance in neglect. CONCLUSIONS: There is substantial involvement of fathers in a subset of this high-risk sample, although more than a quarter of the children lacked a father or father figure. The mere presence of a father did not significantly influence the neglect of the children; rather, the nature of his involvement did. Fathers who felt more effective as parents were less likely to have neglected their children. A greater sense of efficacy may reflect parenting skills and be important in enhancing the contribution of fathers to their children's well-being. Pediatric health care providers can play a valuable role in enhancing the involvement and skills of fathers.

A survey of IgA protease production among clinical isolates of Proteeae.

A collection of 100 strains of Proteeae, in which all species within the tribe were represented, was examined for IgA protease production. The strains were isolated from various clinical specimens from sick and healthy persons in several countries. IgA protease-producing strains were not found amongst species of Providencia and Morganella but were common in Proteus spp. All the strains of P. mirabilis and P. penneri and many of the strains of P. vulgaris examined produced an EDTA-sensitive protease that cleaved the IgA heavy chain outside the hinge region. The proteus enzyme was different in this respect from the EDTA-sensitive, hinge-cutting proteases of other bacteria. The ability to produce IgA protease was unrelated to the O antigenicity, biotype or bacteriocin type of the strain. IgA protease production may be an important virulence mechanism for Proteus strains.