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A Tagliabue

Publications and source records attributed to A Tagliabue.

At least 19 recordsLinked to original sources
PubMed1995

Mapping of receptor binding sites on IL-1 beta by reconstruction of IL-1ra-like domains.

Upon structure comparison between IL-1 beta and its antagonist IL-1ra, single or multiple residues along the IL-1 beta sequence were replaced with the corresponding amino acids present in the IL-1ra protein, in the attempt to identify sites important for receptor binding and for biologic activity on the two molecules. Ten of fifteen mutant proteins had activity comparable to that of wild-type IL-1 beta in three different biologic assays and in receptor binding, indicating that the introduced changes did not influence the functional structure of the protein. Conversely, three mutants (SMIL-9: 127/263 R/T-->W/Y; SMIL-10: 125/127/263/265 T/R/T/Q-->R/W/Y/E; SMIL-15:222/227 I/E-->S/S) showed an increased binding capacity for IL-1RI, not paralleled by increased agonist activity, indicating that the introduced IL-1ra residues could be involved in the nonagonist IL-1RI binding site. On the other hand, two mutants showed diminished binding capacity with concomitant decrease in biologic activity. Both mutants (SMIL-1, five substitutions in the loop 202-214; and SMIL-3, total replacement of the loop 164-173 with the IL-1ra stretch 52-55) included substitutions of residues allegedly important for agonist binding to IL-1RI. Mutant SMIL-3 showed the most profound reduction in binding capacity for IL-1RI (CDw121a) and a more than 1,000-fold reduced biologic activity both in vitro and in vivo, but it retained full capacity of binding to IL-1RII (CDw121b) and acted as a selective antagonist of IL-1RII. From these results the following conclusions can be drawn. IL-1 beta binds to IL-1RI and to IL-1RII through different sites, and the loop 164-173 appears as one of the areas involved in the selective interaction with IL-1RI. Agonist (IL-1 beta) and nonagonist (IL-1ra) binding to IL-1RI occur through distinct sites, with loops 164-173 and 202-214 of IL-1 beta identified as two of the sites selectively involved in agonist binding to the activating receptor.

Animals↗
PubMed1995

Single-cell analysis of macrophage chemotactic protein-1-regulated cytosolic Ca2+ increase in human adherent monocytes.

The increase in intracellular free Ca2+ ([Ca2+]i) associated with interaction of monocyte chemotactic protein-1 (MCP-1) and related chemokines beta with adherent human blood monocytes was investigated at the single-cell level. We used f-MLP as reference chemotactic agent. MCP-1 caused an increase in [Ca2+]i in individual adherent monocytes, with 95% of cells responding to the chemokine at 20 ng/mL. Response to MCP-1 was already detectable at 1 pg/mL, whereas at least 5 ng/mL were required for significant chemotactic response. The kinetics of the increase in [Ca2+]i were considerably different for MCP-1 compared with f-MLP. MCP-1 produced a slow increase of [Ca2+]i that reached a plateau in 5 to 7 minutes. On the other hand, the increase of [Ca2+]i induced by f-MLP appeared to be biphasic, with a fast phase peaking after 5 to 40 seconds followed by a slower wave. Blocking of Ca2+ channels by Ni2+ or Cd2+ and/or chelation of extracellular free Ca2+ considerably reduced but did not abolish response to MCP-1, had no effect on the first wave of [Ca2+]i induced by f-MLP, and completely abrogated the second, slower wave. Thapsigargin, which empties intracellular Ca2+ stores, inhibited f-MLP-induced [Ca2+]i increase but fully blocked the action of MCP-1 only when combined with Ni2+. Thus, increase of [Ca2+]i induced by MCP-1 is apparently due to independent opening of a channel and mobilization from intracellular stores, whereas f-MLP-induced mobilization of Ca2+ from stores causes subsequent opening of a channel. At variance with MCP-1, the related chemokine MCP-2 induced only a low increase of [Ca2+]i in about 40% of adherent monocytes. Inhibition of chemokine-induced increase of [Ca2+]i by cholera or pertussis toxin indicated that MCP-1 and MCP-2 activate monocytes through different intracellular pathways. These results demonstrate at the single-cell level that the mechanisms and dynamics of increased [Ca2+]i are considerably different for f-MLP and chemokines beta. In addition, the [Ca2+]i increase induced by the two related chemokines beta MCP-1 and MCP-2 appears to be differently regulated, suggesting interaction with distinct receptors.

Cadmium↗
PubMed1995

Multi-frequency impedance for the prediction of extracellular water and total body water.

The relationship between total body water (TBW) and extracellular water (ECW), measured by deuterium oxide dilution and bromide dilution respectively, and impedance and impedance index (height2/impedance) at 1, 5, 50 and 100 kHz was studied. After correction for TBW, ECW was correlated only with the impedance index at 1 and 5 kHz. After correction for ECW, TBW was best correlated with the impedance index at 100 kHz. The correlation of body-water compartments with impedance values obtained with modelling programs was lower than with measured impedance values. Prediction formulas for ECW (at 1 and 5 kHz) and TBW (at 50 and 100 kHz) were developed. The prediction errors for ECW and TBW were 1.0 and 1.7 kg respectively (coefficient of variation 5%). The residuals of both ECW and TBW were related to the ECW/TBW value. Application of the prediction formulas in a population, independently measured, revealed a slight overestimation of TBW and ECW, which could be largely explained by differences in the validation group in body-water distribution and in body build. The ratio of impedance at 1 kHz to impedance at 100 kHz was correlated with body-water distribution (ECW/TBW). The relation is however not strong enough to be useful as a predictor. It is concluded that an independent prediction of ECW and TBW, using impedance at low and high frequency respectively, is possible, but that the bias depends on the body-water distribution and body build of the measured subject.

Adult↗
PubMed1995

The reproducibility of subjective appetite scores.

Although subjective appetite scores are widely used, studies on the reproducibility of this method are scarce. In the present study nine healthy, normal weight, young men recorded their subjective appetite sensations before and during 5 h after two different test meals A and B. The subjects tested each meal twice and in randomized order. Visual analogue scale (VAS) scores, 10 cm in length, were used to assess hunger, satiety, fullness, prospective food consumption and palatability of the meals. Plasma glucose and lactate concentrations were determined concomitantly. The repeatability was investigated for fasting values, delta-mean 5 h and mean 5 h values, delta-peak/nadir and peak/nadir values. Although the profiles of the postprandial responses were similar, the coefficients of repeatability (CR = 2SD) on the mean differences were large, ranging from 2.86 to 5.24 cm for fasting scores, 1.36 to 1.88 cm for mean scores, 2.98 to 5.42 cm for delta-mean scores, and 3.16 to 6.44 cm for peak and delta-peak scores. For palatability ratings the CR values varied more, ranging from 2.38 (taste) to 8.70 cm (aftertaste). Part of the difference in satiety ratings could be explained by the differences in palatability ratings. However, the low reproducibility may also be caused by a conditioned satiation or hunger due to the subjects' prior experience of the meals and therefore not just be a reflection of random noise. It is likely, however, that the variation in appetite ratings is due both to methodological day-to-day variation and to biological day-to-day variation in subjective appetite sensations.

Adult↗
PubMed1995

The effect of raw potato starch on energy expenditure and substrate oxidation.

Because resistant starch (RS) is not absorbed as glucose in the small intestine of healthy humans, postprandial thermogenesis should be lower after the intake of RS as compared with digestible starch. To evaluate this hypothesis, we measured 5-h postprandial thermogenesis and substrate oxidation by indirect calorimetry after ingestion of 50 g pregelatinized (0% RS) and 50 g raw potato starch (54% type II RS) in 15 healthy, normal-weight young males. The subjects consumed each starch (mixed in diluted fruit syrup) twice on separate days and in random order. RS intake was followed by lower thermogenesis (46.5 +/- 13.1 compared with 115.4 +/- 10.4 kJ/5 h; P = 0.008), lower glucose oxidation (P < 0.0005), and greater fat oxidation (P = 0.013) than was pregelatinized starch consumption. Our results suggest that RS has no thermogenic effect and that its presence does not influence the size of the thermic response to digestible starch.

Adult↗
PubMed1995

[Symptoms of prostatic hypertrophy: evaluation parameters].

To evaluate BPH symptoms many questionnaires have been proposed. The World Health Organisation Consultation on BPH recommended the "AUA-7 questionnaire" to which and additional on the quality of life was added, creating the "International Prostate Symptom score" (I-PSS). In any case, before proposing a surgical therapy, you have to correlate between symptoms, gland size and urodynamics. The only method to accurately evaluate outflow obstruction is pressure/flow study.

Humans↗
PubMed1994

Resistant starch: the effect on postprandial glycemia, hormonal response, and satiety.

The effect of resistant starch (RS) on postprandial plasma concentrations of glucose, lipids, and hormones, and on subjective satiety and palatability ratings was investigated in 10 healthy, normal-weight, young males. The test meals consisted of 50 g pregelatinized starch (0% RS) (S) or 50 g raw potato starch (54% RS) (R) together with 500 g artificially sweetened syrup. After the R meal postprandial plasma concentrations of glucose, lactate, insulin, gastric inhibitory polypeptide (GIP), glucagon-like peptide-1, and epinephrine were significantly lower compared with after the S meal. Moreover, subjective scores for satiety and fullness were significantly lower after the R meal than after the S meal. Differences in GIP, texture, and palatability may have been involved in these findings. In conclusion, the replacement of digestible starch with RS resulted in significant reductions in postprandial glycemia and insulinemia, and in the subjective sensations of satiety.

Adult↗
PubMed1994

Coffee induced thermogenesis and skin temperature.

The purpose of this study was to investigate the relationship between the changes in mean skin temperature and the energy expenditure induced by drinking coffee containing 4 mg of caffeine/kg body weight. Twelve healthy, weight-stable subjects were studied (five males, seven females; mean age +/- s.d., 25.3 +/- 3.3 years; BMI 22.5 +/- 3.1). Energy expenditure (EE) was measured by open-circuit indirect calorimetry, and skin temperature was determined with thermometric probes applied to the four body regions indicated by Ramanathan (chest, arm, thigh, calf). The calorimetric and thermometric measurements were carried out for 120 min with the participants lying down quietly. A significant correlation was found between the total thermogenic responses (net responses) and the temperature changes between 90 and 120 min from the coffee intake. Multiple regression analysis using mean EE after coffee intake as the dependent variable, and mean skin temperature and body weight as the independent variables yields the following equation: EE (kcal/min/m2) = -1.44 + 0.052 (mean skin temperature) + 0.004 (body weight). (r = 0.71 and P = 0.01) Our results indicate that small interindividual differences in mean skin temperature could explain energy expenditure differences in subjects with the same body weight, body composition and physical activity. This, in turn, could help explain variations in proneness to obesity.

Adult↗
PubMed1992

Interaction of the pertussis toxin peptide containing residues 30-42 with DR1 and the T-cell receptors of 12 human T-cell clones.

The interaction of the immunodominant pertussis toxin peptide containing residues 30-42 (p30-42) with soluble DR1 molecules and the T-cell receptor (TCR) of 12 DR1-restricted human T-cell clones has been analyzed. Peptide analogues of p30-42 containing single alanine substitutions were used in DR1-binding and T-cell proliferation assays to identify the major histocompatibility complex and TCR contact residues. Each T-cell clone was found to recognize p30-42 with a different fine specificity. However, a common core comprising amino acids 33-39 was found to be important for stimulation of all T-cell clones. Within this core two residues, Leu33 and Leu36, interact with the DR1 molecule, whereas Asp34, His35, Thr37, and Arg39 are important for TCR recognition in most of the clones. Computer modeling of the structure of p30-42 showed that an alpha-helical conformation is compatible with the experimental data. The analysis of TCR rearrangement revealed that the peptide was recognized by T-cell clones expressing different variable region alpha (V alpha) and variable region beta (V beta) chains, although a preferential use of V alpha 8-V beta 13 and V alpha 11-V beta 18 combinations was found in clones from the same donor. Understanding the details of the interaction of antigenic peptides with the major histocompatibility complex and TCR molecules should provide the theoretical basis to design T-cell epitopes and obtain more immunogenic vaccines.

Amino Acid Sequence↗
PubMed1992

Binding and internalization of the 163-171 fragment of human IL-1 beta.

The mechanisms of cell association of the human interleukin (IL-1 beta) immunostimulatory fragment 163-171 have been studied. The fragment was able to associate abundantly to both IL-1R- and IL-1R+ cells. Binding was strictly temperature dependent, was not saturable and could be inhibited by excess amounts of unlabelled 163-171 peptide but not by IL-1 beta, suggesting that the 163-171 fragment is not an IL-1R-binding domain of IL-1 beta. The fragment is readily internalized by cells by a cytochalasin-insensitive mechanism and it localizes mainly in the cytoplasm. It is concluded that the active domain 163-171 of IL-1 beta can be taken up by cells through a receptor-independent, temperature-dependent mechanisms and that its ability to activate cellular functions is based on IL-1R-independent intracellular pathways.

Amino Acid Sequence↗
PubMed1992

How reliable is bio-electrical impedance analysis for individual patients?

We performed bio-electrical impedance analysis (BIA) on 38 healthy adults with an initial body mass index (+/- s.d.) of 30 kg/m2 (+/- 5) before and after a low calorie diet. Five weeks later a mean weight loss of 4.2 +/- 2.3 kg was observed. According to BIA, fat-free mass (FFM) decreased in 28 subjects and increased in ten. In four cases the reduction was greater than the weight loss. At the end of the ten week diet regimen all 27 subjects followed up demonstrated a further weight reduction. According to BIA, FFM decreased in 22 subjects, increased in four and did not change in one case. In one case the reduction was greater than the weight loss. Although mean FFM values assessed by skinfolds and BIA were not significantly different before and after the weight loss period, no correlations were found among the individual changes in FFM evaluated by the two methods. Our results suggest that single frequency (50 KHz) impedance must be used with care in clinical conditions.

Adult↗
PubMed1991

Binding of IL-1 beta to IL-1R type II at single cell level.

To gain information on the possible biologic role of IL-1R type II (IL-1RII), expression of the 68-kDa IL-1 binding protein on human lymphoblastoid B cells was investigated at single cell level. Binding of iodinated IL-1 beta was evaluated by autoradiography on cytosmears of IL-1RII positive B cell lines RAJI, the RAJI clone 1H7, and STS 25. Results obtained suggest an heterogeneity of IL-1RII expression within the B cell population, with only 5 to 16% of the cells able to bind IL-1 beta. Up-regulation of IL-1RII expression by dexamethasone, evident in conventional binding assays, was achieved through both increase in the number of IL-1 binding cells (14-30%) and augmentation of receptor density on positive cells, By combining autoradiography with immunocytochemical staining, it could be shown that about 80% of IL-1RII + cells were negative for Ki67, a nuclear antigen expressed from late G1 to M phase. Cell cycle dependent expression of IL-1RII was confirmed on cells enriched in different phases of the cell cycle by counterflow centrifugal elutriation. It is thus proposed that IL-1RII is associated to the cell cycle.

Antigens, CD↗
PubMed1991

Quantitation of biologically active IL-1 by a sensitive assay based on immobilized human IL-1 receptor type II (IL-1RII).

A rapid and sensitive solid-phase radioassay is described for the quantitative detection of human interleukin-1 (IL-1) based on its capability to bind the nitrocellulose-immobilized IL-1 receptor solubilized from plasma membranes of a subclone of the human B cell lymphoma Raji. The assay can detect human IL-1 beta levels as low as 1 X 10(-11) M, both in physiological buffers and in human plasma. Much lower sensitivity was observed for human IL-1 alpha (3.7 X 10(-9) M) and murine IL-1 beta (2 X 10(-9) M). This assay has the advantage to specifically detect only the correctly folded biologically active IL-1. Simple pretreatment procedure that selectively removes IL-1 beta from samples has been devised so that the ratio of the two IL-1s isoforms in the sample can be precisely determined. This assay represents a fast method for the simultaneous-testing of large numbers of biological samples.

Filtration↗
PubMed1991

Lymphokine secretion and cytotoxic activity of human CD4+ T-cell clones against Bordetella pertussis.

Human CD4+ T-cell clones specific for pertussis toxin and other Bordetella pertussis antigens have been tested for their cytotoxic activity, lymphokine production, and capacity to induce immunoglobulin synthesis. Clones specific for the S1 subunit of pertussis toxin were cytotoxic for autologous Epstein-Barr virus-transformed B cells, which had been pulsed with the native antigen, the recombinant S1 subunit of pertussis toxin, or synthetic peptides derived from the S1 amino acid sequence. The killing of antigen-pulsed target cells was class II restricted. All of the T-cell clones produced mostly interleukin-2 and gamma interferon and assisted allogeneic B cells in the production of immunoglobulins M and G but not immunoglobulin E. The potential in vivo role of the cytotoxic activity of these clones is discussed.

B-Lymphocytes↗
PubMed1991

Identification of subregions of Bordetella pertussis filamentous hemagglutinin that stimulate human T-cell responses.

Filamentous hemagglutinin (FHA), a 220-kDa protein that mediates the adhesion of Bordetella pertussis to eukaryotic cells, is a component of acellular vaccines against whooping cough. To identify the subregions of FHA that are immunogenic for T cells, 16 human T-cell clones were raised against purified FHA and tested for the recognition of recombinant and proteolytic fragments. The clones were found to map either in the carboxy-terminal or the amino-terminal part of the FHA molecule, but none of them recognized the central region, which contains a sequence that is homologous to that of the eukaryotic protein fibronectin. These data suggest that subregions of FHA that do not contain sequences that are potentially cross-reactive with self proteins may be sufficient to induce an immune response against the whole protein.

Adhesins, Bacterial↗
PubMed1991

Obesity, diet and body temperature.

Studies conducted thus far on the pathogenesis of obesity have not clearly determined the role of body temperature in the energy balance. In an attempt to explore this relationship further, research has been undertaken, a part of which is represented by the present investigation carried out on 22 adult males: 11 having a BMI less than or equal to 25 (group A) and 11 with BMI greater than 25 (group B). Body temperature was measured by mouth and on the skin surface (in 4 points according to Ramanathan) on 2 consecutive days (every 30 min from noon to 6:30 p.m.), the 1st day with the participants fasting and the 2nd day after consumption of a meal whose energy content represented a part (45%) of the 24-hour energy intake of the subjects, as previously ascertained by a survey of their food consumption. The study was carried out under frequently checked microclimatic conditions. During the week prior to the measurements, the alimentary consumption of the subjects was monitored by direct weighing for 5 consecutive days. A significant rise in body temperature, probably due to dietary thermogenesis, was observed in group A (internal temperature: F = 13.05; skin temperature: F = 6.48) as well as in group B (internal temperature: F = 24.88; skin temperature: F = 5.35) after the meal. However in group B the skin temperature showed a smaller increase than in group A (delta t degree 0.31 vs. 0.49) and an earlier trend towards the basal values (nearly 5 vs. 6 h after the meal). In both groups a decrease in the skin delta t degrees is evident at 3 and 3.5 h, respectively, after the meal, followed by a more remarkable increase in this difference between the 4th and the 5th hour. The meaning of this decrease as well as the difference in body temperature versus caloric intake is discussed. Normal weight individuals demonstrated a positive correlation (r = 0.83) between usual caloric intake per square meter of body surface and fasting internal body temperature. This correlation was not observed in subjects with BMI greater than 25.

Adult↗
PubMed1991

Human T cell immunity against Bordetella pertussis analyzed at clonal level.

Cloned peripheral blood T lymphocytes from an immune donor were grown in interleukin 2 and tested for proliferation in response to inactivated Bordetella species (B. pertussis, B. parapertussis and B. bronchiseptica) and mutants deficient for the expression of virulence-associated antigens. All the T cell clones obtained were CD4+8- and recognized specifically the Bordetella antigens when presented by autologous B cells. On the basis of the responsiveness to the whole inactivated bacteria, it was possible to cluster the twelve clones obtained into four groups with the following specificity: 1) filamentous hemagglutinin (FHA); 2) B. pertussis specific antigens; 3) virulence-associated Bordetella specific antigens; 4) non-virulence-associated Bordetella specific antigens. Employing two new B. pertussis deletion mutants, clone 6 (representative of cluster 1), was found to recognize the C-terminus of FHA. Furthermore, 3 out of 4 clones of cluster 3 were specifically stimulated by the soluble 69,000 M.W. protein from the outer membrane of B. pertussis. Surprisingly, none of the twelve clones obtained by stimulation in vitro with whole inactivated bacteria recognized PT. Thus, PT does not seem to be the most representative antigen on the whole inactivated bacteria. However, when a new generation of clones was obtained using soluble PT as the in vitro stimulus, it was observed that 11 clones of this group recognized this antigen. Furthermore, the majority of them was against the subunit Sl of PT. Therefore, we can conclude that a T cell memory against PT exists in a donor who has had pertussis several years before. In conclusion, these results provide useful information in the attempt to obtain a simplified acellular vaccine for whooping cough.

Antigen-Presenting Cells↗