Isolation of endothelial cells from human first trimester decidua using immunomagnetic beads.

A technique was developed for the isolation of purified endothelial cells from first trimester decidual tissue, using QB End/40-coated magnetic polystyrene beads (Dynabeads). QB End/40 is an endothelial-specific monoclonal antibody which reacts with the coagulation cascade co-factor, thrombomodulin. Small endothelial cell aggregates were released from first trimester decidua by the sequential use of collagenase and trypsin. This dissociation method yielded 15-20% endothelial cells which were further purified to greater than 90% homogeneity by rosetting with QB End/40-Dynabeads. Cultures of purified decidual endothelial cells provide a useful tool for investigating cell-cell interaction in the first trimester placental bed.     

Human trophoblast adhesion to matrix proteins: inhibition and signal transduction

At the time of implantation, the extracellular matrix proteinslaminin and fibronectin are abundant in the decidua and aredistributed pericellularly around each individual stromal cell.First trimester human trophoblast expresses both laminin andfibronectin receptors, specifically the ₁ß_{1 5}ß₁₆ß₁ and ₆ß₄ integrin heterodimers. In thisstudy we have demonstrated that in-vitro adhesion of first trimesterhuman trophoblast to purified extracellular matrix proteinsand to purified decidual stromal cell monolayers can be inhibitedby monoclonal antibodies directed against appropriate integrinsubunits and by synthetic peptides containing an arginine-glycine-asparticacid sequence. Monoclonal antibodies (mAbs) to the ₅ and ß₁integrin subunits and a synthetic peptide significantly inhibitedadhesion to fibronectin. Binding of trophoblast to laminin wasblocked with mAbs to the ₆ and ß₁ but not ₁ and ß₄integrinsubunits. Similarly, integrin-mediated adhesion to monolayersof decidual stromal cells could be blocked with mAbs to the₅, ₆, ß₆ and ß₄ integrin subunits. Integrin-mediatedsignal transduction in normal and malignant trophoblast wasinvestigated by Western blotting. A 115 kDa protein was themajor tyrosine phosphorylated protein detected in trophoblastafter binding to laminin or fibronectin. The profile of tyrosinephosphorylated proteins differed for malignant trophoblast. integrins/matrix/signal transduction/trophoblast     

Separation anxiety: Extreme dehiscence of a mitral annuloplasty ring

We report a patient admitted with acute pulmonary edema 3 months after mitral valve repair, with no history of inter‐current febrile illness. Transesophageal echocardiography (TEE) demonstrated severe mitral regurgitation (MR) and an abnormally positioned annuloplasty ring, suggestive of dehiscence. The extreme extent of ring dehiscence was visualized on 3‐dimensional TEE (3D), with near‐complete separation of the ring. Strept.Mitis and Cristatus were isolated from the ring following redo mitral valve surgery, confirming endocarditis as the mechanism for dehiscence. This report highlights the additive role and superior ability of 3D TEE in the identification and anatomic delineation of mitral ring dehiscence.     

Endogenous endothelial nitric oxide synthase-derived nitric oxide is a physiological regulator of myocardial oxygen consumption

Our objective was to determine the precise role of endothelial nitric oxide synthase (eNOS) as a modulator of cardiac O2 consumption and to further examine the role of nitric oxide (NO) in the control of mitochondrial respiration. Left ventricle O2 consumption in mice with defects in the expression of eNOS [eNOS (-/-)] and inducible NOS [iNOS (-/-)] was measured with a Clark-type O2 electrode. The rate of decreases in O2 concentration was expressed as a percentage of the baseline. Baseline O2 consumption was not significantly different between groups of mice. Bradykinin (10(-4) mol/L) induced significant decreases in O2 consumption in tissues taken from iNOS (-/-) (-28+/-4%), wild-type eNOS (+/+) (-22+/-4%), and heterozygous eNOS(+/-) (-22+/-5%) but not homozygous eNOS (-/-) (-3+/-4%) mice. Responses to bradykinin in iNOS (-/-) and both wild-type and heterozygous eNOS mice were attenuated after NOS blockade with N-nitro-L-arginine methyl ester (L-NAME) (-2+/-5%, -3+/-2%, and -6+/-5%, respectively, P<0.05). In contrast, S-nitroso-N-acetyl-penicillamine (SNAP, 10(-4) mol/L), which releases NO spontaneously, induced decreases in myocardial O2 consumption in all groups of mice, and such responses were not affected by L-NAME. In addition, pretreatment with bacterial endotoxin elicited a reduction in basal O2 consumption in tissues taken from normal but not iNOS (-/-)-deficient mice. Our results indicate that the pivotal role of eNOS in the control of myocardial O2 consumption and modulation of mitochondrial respiration by NO may have an important role in pathological conditions such as endotoxemia in which the production of NO is altered.     

A Clinico-Pathological Analysis of Seventeen Cases of Non-Hodgkin'S Lymphoma Involving the Orbit

Seventeen patients with non-Hodgkin's lymphoma involving the orbit were studied. There were 12 males and 5 females and their median age was 53 years. Three cases were classified as stage I, two as stage II and 12 as stage IV. the histology was small lymphocytic type in 4 of the 5 stage I and II cases, and immunophenotyping, confirming their B-cell origin, was performed in two of these cases. All stage I and II patients responded favourably to local radiotherapy alone. the 12 stage IV patients had a marked male predominance and more commonly an intermediate grade histology and T-immunophenotype. Several of the orbital lesions seemed to represent extension of nasal lymphomas. the stage IV patients had a poorer survival than the stage I and II patients but the difference was not statistically significant, probably due to the small number of patients.