Sphingosine-1-phosphate as a ligand for the G protein-coupled receptor EDG-1

The sphingolipid metabolite sphingosine-1-phosphate (SPP) has been implicated as a second messenger in cell proliferation and survival. However, many of its biological effects are due to binding to unidentified receptors on the cell surface. SPP activated the heterotrimeric guanine nucleotide binding protein (G protein)-coupled orphan receptor EDG-1, originally cloned as Endothelial Differentiation Gene-1. EDG-1 bound SPP with high affinity (dissociation constant = 8.1 nM) and high specificity. Overexpression of EDG-1 induced exaggerated cell-cell aggregation, enhanced expression of cadherins, and formation of well-developed adherens junctions in a manner dependent on SPP and the small guanine nucleotide binding protein Rho.     

Macrophage migration inhibitory factor production by Leydig cells: evidence for a role in the regulation of testicular function

Macrophage migration inhibitory factor (MIF), described originally as a product of activated T lymphocytes, recently has been found to be released by monocytes/macrophages and the anterior pituitary gland. Immunohistochemical studies of the adult rat testis using an affinity-purified polyclonal antimurine MIF antibody demonstrated strong staining for MIF in Leydig cells and their putative precursors. Peritubular myoid cells and the seminiferous epithelium were negative for MIF staining; however, a weak reaction around the heads of elongated spermatids also was observed. The expression of MIF messenger RNA and protein in whole rat testis was demonstrated by Northern blot and Western blot analyses, respectively. Both MIF messenger RNA and protein immunoreactivity in Leydig cells was observed in testes obtained from long term hypophysectomized rats. Significant concentrations of intracellular MIF were detected in lysates of the TM3 Leydig cell line (7.23 +/- 2.6 pg/microgram protein), and testicular interstitial fluid contained 14.7 +/- 1.6 ng/ml MIF protein, as measured by MIF-specific enzyme-linked immunosorbent assay. To gain insight into the possible biological role of MIF in the testis, cultures of adult rat seminiferous tubules and purified Leydig cells were incubated together with recombinant murine MIF (rMIF). Neither rMIF (50 ng/ml) nor a neutralizing anti-MIF antiserum was found to affect basal or LH-stimulated Leydig cell steroidogenesis in vitro. However, a dose-dependent decrease in the secretion of inhibin by the seminiferous tubules was observed at rMIF concentrations ranging from 10-100 ng/ml. Taken together, these data indicate that Leydig cells produce MIF in vivo and suggest an important regulatory role for this newly discovered mediator of testicular function.     

papG alleles among Escherichia coli strains causing urosepsis: associations with other bacterial characteristics and host compromise

Alleles I, II, and III of the P adhesin gene papG were sought by PCR among 75 Escherichia coli blood isolates from adults with urosepsis, and the papG genotype was compared with associated bacterial characteristics and host compromise status. Allele II predominated over allele III in the total population (71% of the strains versus 7% for allele III; P < 0.01). Allele I was not encountered. In comparison with allele II, allele III was significantly associated with the presence of hly and the absence of iuc (which encode hemolysin and aerobactin biosynthesis), nonagglutination of digalactoside-coated beads, absence of aerobactin production, membership in serogroups O6 and O18, and host compromise, particularly cancer and upper urinary tract abnormalities.     

The clinical and physiological effect of whole-lung lavage in pulmonary alveolar proteinosis: a ten-year experience

We have utilized whole-lung lavage in the successful treatment of 18 patients with pulmonary alveolar proteinosis. Our ten-year experience includes serial evaluations of patients with disabling lung dysfunction who had a total of 49 whole-lung lavages under general anesthesia. Clinical and physiological responses were documented both before and after each lavage. There were no complications or deaths. All patients were radiographically, physiologically, and symptomatically improved within hours after the procedures. Five patients required from two to four repeat lavages one to three years later. The treatment of this disorder has included a wide variety of techniques. We attribute our results to the use of a lung lavage technique that includes: (1) unilateral whole-lung lavages at two to three day intervals; (2) isotonic saline as the lavage solution; (3) use of a mechanical chest percussor during lavage; and (4) measuring the total thoracic compliance of each side in the immediate postlavage period as a guide for extubation. We conclude that whole-lung lavage is a safe, highly effective, repetitively applicable treatment for pulmonary alveolar proteinosis.     

Effect of Grinding on Flaw Geometry and Fracture of Glass

Fracture mechanics is combined with fracture surface analysis to analyze brittle failure of glass bars which were tested relative to the direction of grinding. Grinding essentially produces two sets of flaws from which failure occurs. In the most severe set, formed basically parallel to the grinding direction, the ratio of the average depth ( a ) to the half-width ( b ) is 0.5. In the less severe set, formed perpendicular to the grinding direction, the average a / b ratio is 1.6. In both sets the most severe flaws are generally associated with a particularly deep grinding groove or gouge. The strength reduction resulting from testing perpendicular to the grinding direction results from the larger flaw size and slightly higher stress-intensity factor resulting from the greater ellipticity of the flaws formed parallel to the grinding grooves and perpendicular to the tensile axis. Detailed analysis of these 2 sets of flaws causing failure of appropriately oriented specimens shows that (1) the fracture mirror radius, r , occurs at a constant stress-intensity level independent of flaw geometry; (2) unsymmetric fracture mirrors result from unsymmetric, irregular flaws leading to unsymmetric stress-intensity distributions; (3) is constant for semielliptical flaws; and (4) fracture energy calculated from an expression including mirror constants, the flaw-to-mirror size ratio, and the flaw geometry agrees with measured values over a wide range of a / b values.     

Amplified migration inhibition effect

Upon exposure to specific antigen in tissue culture, sensitive lymphocytes released macrophage migration inhibition factor and other lymphokines into the supernatant culture medium. Migration of peritoneal macrophages from nonsensitive animals was inhibited in the presence of such supernatants. However, with previous techniques it was found that an inhibitory effect was present at only low low titers (less than 10(2)). It is therfore of great interest that by increasing cellular density, the total number of cells being kept constant, inhibitory activity can be amplified by a factor as great as 10(10). This amplification was observed only when lymphocytes and macrophages were loosely packed, as by spontaneous sedimentation in a conical test tube. The effect was abolished by dispersing the cell suspension in a flat-bottomed flask or, alternatively, by shaking the test tube so that intimate prolonged intercellular contact was prevented.