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Biomedical subjects

J R Wright

Publications and source records attributed to J R Wright.

At least 73 records · Page 4Linked to original sources
PubMed1999

Surfactant protein A enhances alveolar macrophage phagocytosis of a live, mucoid strain of P. aeruginosa.

In this study, we investigate the interaction between surfactant protein A (SP-A) and a live, mucoid strain of Pseudomonas aeruginosa and identify a mechanism of clearance of this organism by alveolar macrophages. (125)I-labeled SP-A bound live, but not heat-killed, P. aeruginosa organisms in a concentration-dependent manner. Unlabeled SP-A bound live bacteria, protein isolated from whole organisms, and specific proteins of the P. aeruginosa outer membrane. The binding of SP-A to P. aeruginosa and outer membrane components was inhibited by either EDTA or mannose. Phagocytosis assays with fluorescent microscopy demonstrated that the percentage of macrophages with internalized FITC-labeled P. aeruginosa was increased 1.8-fold (19 vs. 35%) by pretreating the live bacteria with SP-A. This finding was confirmed by direct visualization of ingested bacteria by electron microscopy. Adhering macrophages to SP-A-coated surfaces attenuated the increased uptake of P. aeruginosa pretreated with SP-A, suggesting that SP-A acts as an opsonin to stimulate macrophage phagocytosis of this strain of P. aeruginosa.

Animals↗
PubMed1999

Effects of muscle perfusion pressure on fatigue and systemic arterial pressure in human subjects.

The effects of changes in arterial perfusion across the physiological range on the fatigue of a working human hand muscle were studied in seven normal subjects. With the hand above heart level, subjects made repeated isometric contractions of the adductor pollicis muscle at 50% of maximal voluntary contraction in a 6-s on, 4-s off cycle. To assess fatigue, a maximal isometric twitch was elicited in each "off" period by electrical stimulation of the ulnar nerve. The experiment was repeated at least 2 days later with the hand at heart level. Five subjects showed faster fatigue with the arm elevated, and two subjects showed little difference in fatigue for the two conditions. Central blood pressure rose in proportion to fatigue for the subjects overall and returned quickly to its initial level afterwards. We conclude that human muscle fatigue can be increased by physiological reductions in perfusion pressure. Central blood pressure increases as the muscle fatigues, a response that may partially offset declining muscle performance.

Adult↗
PubMed1999

Serial changes in surfactant-associated proteins in lung and serum before and after onset of ARDS.

The goal of this study was to determine the changes that occur in surfactant-associated proteins in bronchoalveolar lavage fluid (BAL) and serum of patients at risk for ARDS and during the course of ARDS. We found that the concentrations of SP-A and SP-B were low in the BAL of patients at risk for ARDS before the onset of clinically defined lung injury, whereas the concentration of SP-D was normal. In patients with established ARDS, BAL SP-A and SP-B concentrations were low during the entire 14-d observation period, but the median SP-D concentrations remained in the normal range. Immunoreactive SP-A and SP-D were not increased in the serum of patients at risk for ARDS, but both increased after the onset of ARDS to a maximum on Day 3 and remained elevated for as long as 14 d. The BAL SP-A concentrations were significantly lower in at-risk patients who developed ARDS, and no patient with a BAL SP-A concentration greater than 1.2 microg/ml developed ARDS. On Days 1 and 3 of ARDS, the BAL SP-D concentration was significantly lower in patients who died, and the BAL SP-D concentration was significantly related to the PI(O(2))/FI(O(2)) ratio. Thus, surfactant protein abnormalities occur before and after the onset of ARDS, and the responses of SP-A, SP-B, and SP-D differ in important ways. The BAL SP-A and SP-D measurements can be used to classify patients as high or low risk for progression to ARDS and/or death after the onset of ARDS. Strategies to increase these surfactant proteins in the lungs of patients with ARDS could be useful to modify the onset or the course of ARDS.

APACHE↗
PubMed1999

Surfactant protein-A binds group B streptococcus enhancing phagocytosis and clearance from lungs of surfactant protein-A-deficient mice.

Surfactant protein-A (SP-A) gene-targeted mice clear group B streptococcus (GBS) from the lungs at a slower rate than wild-type mice. To determine mechanisms by which SP-A enhances pulmonary clearance of GBS, the role of SP-A in binding and phagocytosis of GBS was assessed in SP-A (-/-) mice infected with GBS in the presence and absence of exogenous SP-A. Coadministration of GBS with exogenous SP-A decreased GBS colony counts in lung homogenates of SP-A (-/-) mice. SP-A bound to GBS in a calcium-dependent manner. Although pulmonary infiltration with macrophages was not altered in SP-A (-/-) versus wild-type mice after GBS infection, the number of alveolar macrophages with phagocytosed bacteria was lower in the SP-A (-/-) mice than in the wild-type mice. When SP-A was coadministered with GBS, phagocytosis was significantly increased. Oxygen radical production by alveolar macrophages from SP-A (-/-) mice infected with GBS was decreased compared with wild-type controls and was increased when SP-A (-/-) mice were infected in the presence of exogenous SP-A. Superoxide (SO) radical generation was deficient in macrophages from SP-A (-/-) mice. SP-A plays an important role in GBS clearance in vivo, mediated in part by binding to and enhancing GBS phagocytosis and by increasing SO production by alveolar macrophages.

Animals↗
PubMed1999

Glycoprotein-340 binds surfactant protein-A (SP-A) and stimulates alveolar macrophage migration in an SP-A-independent manner.

Glycoprotein-340 (gp-340) was first identified as a surfactant protein (SP)-D-binding molecule purified from lung lavage of patients with alveolar proteinosis (Holmskov, et al., J. Biol. Chem. 1997;272:13743). In purifying SP-A from proteinosis lavage, we isolated a protein that copurifies with SP-A and SP-D and that was later found by protein sequencing to be gp-340. We have shown that soluble gp-340 binds SP-A in a calcium-dependent manner independent of the lectin activity of SP-A. To examine the functional significance of this interaction, we tested the ability of soluble gp-340 to block SP-A binding to and stimulation of the chemotaxis of alveolar macrophages. We found that gp-340 does not affect the binding of SP-A to alveolar macrophages over a wide range of SP-A concentrations, nor does it inhibit the ability of SP-A to stimulate macrophage chemotaxis. We also found that gp-340 alone stimulates the random migration (chemokinesis) of alveolar macrophages in a manner independent of SP-A-stimulated chemotaxis. These results suggest that gp-340 is not a cell-surface receptor necessary for SP-A stimulation of chemotaxis, and show that gp-340 can directly affect macrophage function.

Amino Acid Sequence↗
PubMed1999

Surfactant protein D stimulates phagocytosis of Pseudomonas aeruginosa by alveolar macrophages.

Surfactant protein (SP)-D is an oligomeric glycoprotein belonging to the family of collagen-like lectins known as collectins, which have previously been shown to stimulate phagocytosis and other immune cell functions. The hypothesis investigated in this study was that SP-D would stimulate the phagocytosis of an important pulmonary pathogen, Pseudomonas aeruginosa. SP-D, isolated from the lavage fluid of silica-treated rats, significantly enhanced the uptake of three of six strains of P. aeruginosa by rat alveolar macrophages as analyzed by both fluorescence and electron microscopy. SP-D had only minimal effects on phagocytosis of Haemophilus influenzae. SP-D bound to live P. aeruginosa, and binding was inhibited by chelation of calcium and by a competing saccharide, inositol. In vitro killing assays demonstrated that macrophage-mediated killing of one of the mucoid strains of P. aeruginosa was modestly enhanced by SP-D. P. aeruginosa was not measurably aggregated by SP-D either macroscopically or microscopically. Further, SP-D does not appear to act as an activation ligand because adherence of macrophages to SP-D- coated slides did not stimulate the uptake of P. aeruginosa. These findings suggest that SP-D may be important in controlling the pathogenesis of P. aeruginosa in the lung.

Animals↗
PubMed1999

Surfactant protein A suppresses reactive nitrogen intermediates by alveolar macrophages in response to Mycobacterium tuberculosis.

Mycobacterium tuberculosis attaches to, enters, and replicates within alveolar macrophages (AMs). Our previous studies suggest that surfactant protein A (SP-A) can act as a ligand in the attachment of M. tuberculosis to AMs. Reactive nitrogen intermediates (RNIs) play a significant role in the killing of mycobacteria. We have demonstrated that RNI levels generated by AMs were significantly increased when interferon-gamma-primed AMs were incubated with M. tuberculosis. However, the RNI levels were significantly suppressed in the presence of SP-A (10 microg/ml). The specificity of SP-A's effect was demonstrated by the use of F(ab')2 fragments of anti-SP-A monoclonal antibodies and by the use of mannosyl-BSA, which blocked the suppression of RNI levels by SP-A. Furthermore, incubation of deglycosylated SP-A with M. tuberculosis failed to suppress RNI by AMs, suggesting that the oligosaccharide component of SP-A, which binds to M. tuberculosis, is necessary for this effect. These results show that SP-A-mediated binding of M. tuberculosis to AMs significantly decreased RNI levels, suggesting that this may be one mechanism by which M. tuberculosis diminishes the cytotoxic response of activated AMs.

Animals↗
PubMed1999

Comparison of test interval and best prediction methods for estimation of lactation yield from monthly, a.m.-p.m., and trimonthly testing.

A method with best prediction properties that condenses information from all test days into measures of lactation yield and persistency has been proposed as a possible replacement for the test interval method and projection factors. The proposed method uses previously established correlations between individual test days and includes inversion of a matrix for each lactation. Milk weights that were representative of monthly, a.m.-p.m., and trimonthly test plans were examined to compare the accuracy of best prediction and test interval methods for estimating lactation yield. Individual milk weights or daily yields of 658 Canadian cows in 17 herds were selected to correspond to test intervals for 100,000 US cows. For a.m.-p.m. testing, the initial milk weight that was credited was selected randomly from the a.m. or p.m. milking and was alternated thereafter. Trimonthly credits were from one of the first three designated test day weights, selected randomly, and each third designated test weight thereafter. Correlations between 305-d actual lactation yield and lactation estimates by the test interval method were 0.97, 0.96, and 0.93 for monthly, a.m.-p.m., and trimonthly testing, respectively. Corresponding correlations for the best prediction method were 0.97, 0.97, and 0.93. Standard deviations of differences between estimated and 305-d actual yields for monthly, a.m.-p.m., and trimonthly testing were 373, 400, and 546 kg, respectively, for best prediction regressed on herd mean, which was a reduction in estimation error of 4, 6, and 10% over the test interval method. The advantage of best prediction was moderate if two milk weights were recorded monthly and was larger if testing was less frequent. Advantages also were found for fat and protein yields estimated by multitrait best prediction for records with reduced component sampling.

Animals↗
PubMed1999

Mathematical representations of correlations among yield traits and somatic cell score on test day.

Prediction of lactation yields and accuracies of yields for use in genetic evaluation can be improved by including information from test day correlations, especially for milk recording plans that vary in the numbers of milk weights recorded and component samples taken. Daily milk weights for 658 lactations of Canadian cows and monthly test records of milk, fat, and protein yields and somatic cell scores for 500,000 lactations of US cows were used to estimate phenotypic correlations between test days within herd-year. Correlations between daily yields for a designated interval between test days generally were highest for midlactation and were lowest for early and late lactation. Regression (two linear, two quadratic, and interaction effects) on mean DIM and interval between test days predicted correlations with a squared correlation of 0.94 for daily milk yields. Similar relationships were found for US monthly data. Variation in sampling was reduced, computer memory was minimized, and positive definiteness was guaranteed by fitting regressions on simply defined sources of correlation. An autoregressive matrix represented the within-trait correlations very well. The equations developed could be used to derive covariances and, subsequently, to estimate lactation yields and accuracies from combinations of individual daily milk, fat, and protein yields and somatic cell score.

Animals↗
PubMed1998

Interactions of surfactant protein A with epithelial cells and phagocytes.

Surfactant protein A (SP-A) has been shown to bind to and regulate the functions of both alveolar type II cells and immune cells including alveolar macrophages. The interaction of SP-A with type II cells has been shown in vitro to inhibit lipid secretion and to promote the uptake of lipid by these cells and these observations led to the hypothesis that SP-A plays an important role in regulating surfactant turnover and metabolism. The finding that mice made deficient in SP-A by homologous recombination (SP-A -/- mice) have relatively normal surfactant pool sizes has raised the possibility that either redundant mechanisms function in vivo to keep pool sizes normal in the absence of SP-A or that the in vitro findings are not significant in the context of the whole, unstressed animal. The interaction of SP-A with immune cells has been shown to affect a variety of responses which, in general, function to promote host defense against infection. Although SP-A receptors have been identified, additional studies will be required to elucidate the mechanism of interaction of SP-A with these cells and the relative importance of the different receptors in SP-A mediated regulation of cell function.

Epithelial Cells↗
PubMed1998

Recombinant rat surfactant-associated protein D inhibits human T lymphocyte proliferation and IL-2 production.

Components of the airspace-lining material may contribute to the local regulation of immune function within the lung. We report here that recombinant rat pulmonary surfactant-associated protein D (SP-D) inhibits the lectin- and anti-CD3-stimulated proliferation of human PBMCs. Inhibition was associated with a decreased production of IL-2, and the addition of human rIL-2 blocked the inhibitory action of SP-D. These effects were not inhibited by maltose, indicating that the inhibitory activity was not dependent upon the lectin activity of SP-D. Studies employing mutant SP-D lacking N-linked sugars or defective in multimerization further indicated that inhibition was not dependent upon cellular interactions with the N-linked oligosaccharide on SP-D or the oligomerization of trimeric SP-D subunits. Although a peptide containing an inverted DGR showed similar IL-2-dependent effects on anti-CD3-stimulated proliferation, deletion of the conserved DGRDGR sequence near the amino-terminal end of the collagen domain did not decrease the suppressive activity of SP-D. We hypothesize that SP-D can dampen lymphocyte responses to exogenous stimuli and protect the lung against collateral immune-mediated damage.

Amino Acid Sequence↗
PubMed1998

GLUT-4 Deficiency and severe peripheral resistance to insulin in the teleost fish tilapia.

Teleost fish, in general, are glucose intolerant; this trait has been attributed to piscine islets secreting insulin primary in response to amino acid secretogogues rather than glucose. However, pancreatic islet from the teleost fish tilapia, when transplanted into diabetic nude mice, were glucose responsive even though tilapia were severely glucose intolerant. This suggested a strong peripheral resistance to the glucostatic effects of insulin. Using Western blotting with polyclonal antibodies as well as Northern analysis for mRNA, tilapia tissues were found to be devoid of GLUT-4, the insulin-sensitive glucose transporter responsible for the hypoglycemic effect of insulin in mammals. The absence of GLUT-4 in peripheral tissues may explain why tilapia, and possibly other teleost fish, are severely glucose intolerant. This suggests that tilapia islets have evolved along mammalian lines to be glucose sensitive while tilapia peripheral tissue have diverged widely. Using the same methods, tilapia were found to have a very limited tissue distribution of the insulin-independent glucose transporter, GLUT-1, which is responsible for basal glucose transport in mammalian cells. It is suggested that tilapia provide a naturally occurring GLUT-4 knockout model.

Adipose Tissue↗
PubMed1998

Cloning, sequencing and characterization of the tilapia insulin gene.

Using degenerate primers based on insulin sequences from other organisms, we report the cloning of the complete tilapia (Oreochromis niloticus) insulin gene. Using nested primers and a cassette ligation strategy we have also cloned 932 base pairs (bp) of 5' flanking and 1152 bp of 3' flanking sequence. The tilapia insulin gene has the similar three exon (one untranslated), two intron distribution found in all insulin genes sequenced to date. However, intron 1 is unique in having a smaller size (73 bp) than found in other organisms. 5' RNA extension revealed the presence of two potential transcriptional start sites. A perfect TATA box is located at -30 bp from the first transcriptional start site. Interestingly, the 5' upstream region contains a microsatellite close to the same position of a unique minisatellite found only in humans and primates. The upstream region also contains several potential control elements to regulate insulin expression that are found in mammalian insulin genes.

Amino Acid Sequence↗
PubMed1998

Degradation of surfactant protein D by alveolar macrophages.

Surfactant protein (SP) D is a pulmonary surfactant-associated protein that may function in lung host defense. SP-D is produced by alveolar type II cells and nonciliated bronchiolar epithelial (Clara) cells of the airway and is secreted into the air space. Here we investigated whether alveolar macrophages degraded SP-D in vitro. We also examined the effects of SP-A and lipids on SP-D metabolism. The results showed that alveolar macrophages bound and degraded SP-D in a time- and temperature-dependent fashion. After 100 min of incubation, the formation of trichloroacetic acid-soluble degradation products increased 4-fold in the medium and 30-fold in the cells. The degradation of SP-D was via a cell-associated process because SP-D was not degraded when incubated in medium previously conditioned by alveolar macrophages. Gel autoradiography of cell lysate samples after incubation with 125I-labeled SP-D demonstrated an increase in degradation products, further confirming the degradation of SP-D by alveolar macrophages. In addition, the degradation of SP-D was not affected by coincubation with SP-A or surfactant-like liposomes containing either phosphatidylglycerol or phosphatidylinositol. In conclusion, alveolar macrophages rapidly degrade SP-D and may play an important role in SP-D turnover and clearance.

Animals↗