Alterations in the fraction of oxygen in inspired gas affect rates of renal prostaglandin E2 synthesis in anesthetized dogs

Summary Reductions of oxygen in inspired gas from 20% to 15%, in anesthetized dogs, reduced arterial PO₂ and increased the renal efflux of PGE₂ but not PGF_2a . Renal blood flow, blood pressure, plasma renin activity as well as arterial pH and PCO₂ were unaffected. PGs may mediate the renal hemodynamic or excretory consequences of alterations in PO₂. In addition, minor variations in PO₂ might account, in part, for the variable renal venous PGE₂ concentrations reported under basal conditions.Authentic prostaglandins were supplied by Dr John E. Pike of the Upjohn Company. This work was supported by grants from the Veterans Administration, the Southern Medical Association and U.S. Public Health Service. Dr. Brash is an American Heart Association, Missouri Affiliate Fellow.     

Arterial and mixed venous blood gases following DNP infusions in rabbits

Summary Hypocapnia and respiratory alkalosis was found in arterial and mixed venous blood after i.v. administration of DNP. Stimulation of ventilation resulted in steady PO₂, when PvO₂ decreased, and seemed to be independent on CO₂.     

Effects of CO2 exposure on distribution of various forms of iron and copper in guinea-pig tissues

Summary The effects on iron and copper distribution and metabolism of exposure to high levels of CO₂ were studied in the guinea-pig. Mature, male animals were placed in an atmosphere of 15% CO₂, 21% O₂ (balance N₂), and sacrificed from 1 h to 1 week thereafter. Total iron and copper concentrations of blood, liver, spleen and bone, as well as concentrations of heme and ferritin iron, were measured together with blood hematocrit, reticulocytes, plasma hemoglobin, plasma ceruloplasmin and copper concentrations. The results show clearly that rapid and sustained red cell damage or hemolysis ensued several h from the start of CO₂ treatment. This resulted in loss of iron and copper from the blood, an influx of both elements into liver, spleen and bone, and a rise in plasma ceruloplasmin. Influx of iron into liver and spleen caused an accumulation of ferritin, the main site for iron storage in cells. Following the effect on red cells, there was an accumulation of heme iron, and a decreased hematocrit, best explained by a depressed activity of the reticuloendothelial and erythropoietic systems. A period of adaptation succeeded these events, in which all blood parameters and most tissue values returned to normal, despite the continuing presence of high CO₂. The only changes not reversed were the elevations in liver, spleen and bone iron stores. These remained high, with a net accumulation of >2 mg iron, or 3–4 times more than originally present. The results indicate that at least in the guinea-pig, high CO₂ exposure results in red cell damage and other events leading to an accumulation of additional iron in the body; also, that iron accumulated as ferritin and hemosiderin in liver and spleen may not be readily available to restore blood hemoglobin concentrations on an acute basis.Acknowledgments. We gratefully acknowledge the technical assistance of Joan R. Moor and Lakshmi Vulimiri with these studies, and the support of Grants No. 17249 and HL22410 from the U.S. Public Health Service.     

Natural killer-like activity in human cultured lymphoid cells propagated in the presence of interleukin-2: acquired resistance to prostaglandin E2- or dexamethasone-mediated suppression

Summary The cytotoxic activity of human peripheral blood lymphocytes against the natural killer-sensitive target K562 was suppressed both by prostaglandin E₂ and dexamethasone. On the other hand, cultured lymphoid cells propagated in the presence of interleukin-2 showed strong cytotoxic reactivity against K562 targets, and were resistant to prostaglandin E₂- or dexamethasone-mediated suppression.     

IP<Subscript>3</Subscript> receptor signaling and endothelial barrier function

The endothelium, a monolayer of endothelial cells lining vessel walls, maintains tissue-fluid homeostasis by restricting the passage of the plasma proteins and blood cells into the interstitium. The ion Ca²⁺, a ubiquitous secondary messenger, initiates signal transduction events in endothelial cells that is critical to control of vascular tone and endothelial permeability. The ion Ca²⁺ is stored inside the intracellular organelles and released into the cytosol in response to environmental cues. The inositol 1,4,5-trisphosphate (IP₃) messenger facilitates Ca²⁺ release through IP₃ receptors which are Ca²⁺-selective intracellular channels located within the membrane of the endoplasmic reticulum. Binding of IP₃ to the IP₃Rs initiates assembly of IP₃R clusters, a key event responsible for amplification of Ca²⁺ signals in endothelial cells. This review discusses emerging concepts related to architecture and dynamics of IP₃R clusters, and their specific role in propagation of Ca²⁺ signals in endothelial cells.     

Über die Wirkung blutzuckersenkender Sulfonylharnstoffe auf das isolierte Rattenzwerchfell

Summary The action of blood sugar depressing sulfonylureas on glucose and oxygen uptake, as well as on glycogen content and formation of C¹⁴O₂ from uniformly labelled C¹⁴-glucose was investigated in rat hemidiaphragms incubated in phosphate buffer. The following results were obtained: (1) Tolbutamide and Carbutamide increased the glucose uptake. (2) Tolbutamide decreased the glycogen-content. (3) Oxygen uptake as well as formation of C¹⁴O₂ were increased by Tolbutamide. (4) The action of Tolbutamide and insuline was equal with respect to glucose uptake but different as regarding the glycogen content, oxygen uptake and CO₂-formation.It is concluded that sulfonylureas increase glucose oxidation in the rat hemidiaphragm probably without increasing insulin sensitivity. To our knowledge this mechanism of action has hitherto not been described.     

A VEGF-A splice variant defective for heparan sulfate and neuropilin-1 binding shows attenuated signaling through VEGFR-2

The development of functional blood and lymphatic vessels requires spatio-temporal coordination of the production and release of growth factors such as vascular endothelial growth factors (VEGFs). VEGF family proteins are produced in multiple isoforms with distinct biological properties and bind to three types of VEGF receptors. A VEGF-A splice variant, VEGF-A₁₆₅b, has recently been isolated from kidney epithelial cells. This variant is identical to VEGF-A₁₆₅ except for the last six amino acids encoded by an alternative exon. VEGF-A₁₆₅b and VEGF-A₁₆₅ bind VEGF receptors 1 and 2 with similar affinity. VEGF-A₁₆₅b elicits drastically reduced activity in angiogenesis assays and even counteracts signaling by VEGF-A₁₆₅. VEGF-A₁₆₅b weakly binds to heparan sulfate and does not interact with neuropilin-1, a coreceptor for VEGF receptor 2. To determine the molecular basis for altered signaling by VEGF-A₁₆₅b we measured VEGF receptor 2 and ERK kinase activity in endothelial cells in culture. VEGF-A₁₆₅ induced strong and sustained activation of VEGF receptor 2 and ERK-1 and −2, while activation by VEGF-A₁₆₅b was only weak and transient. Taken together these data show that VEGF-A₁₆₅b has attenuated signaling potential through VEGF receptor 2 defining this new member of the VEGF family as a partial receptor agonist. Received 31 May 2006; received after revision 26 June 2006; accepted 14 July 2006     

Effect of 15(R)-15-methyl-prostaglandin E2 on iron absorption in the rat

Summary The administration of 15(R)-15-methyl prostaglandin E₂ (15(R)-15-M-PGE₂) in vivo significantly diminished the uptake of⁵⁹Fe into blood, spleen, liver, femur and dried intestine of rats, whereas acetylsalicylic acid (ASA) increased the counts significantly. This effect of ASA was counteracted by 15(R)-15-M-PGE₂. It is suggested that prostaglandins (PGs) might play an important role in inhibiting iron absorption at the intestinal level.This work was supported by grant No.6638 from CONICET (Argentina). The technical assistance of Mrs María E. Castro and Norma Rizzo is gratefully acknowleged.     

A novel interplay between membrane and nuclear melatonin receptors in human lymphocytes: significance in IL-2 production

Human lymphocyte melatonin, through membrane and nuclear receptors binding, acts as an activator in IL-2 production. Antagonism of membrane melatonin receptors using luzindole exacerbates the drop of the IL-2 production induced by PGE₂ in peripheral blood mononuclear and Jurkat cells. This paper studies the melatonin membrane and nuclear receptors interplay in PGE₂-diminished IL-2 production. The decrease in IL-2 production after PGE₂ and/or luzindole administration correlated with downregulation in the nuclear receptor RORα. We also highlighted a role of cAMP in the pathway, because forskolin mimicked the effects of luzindole and/or PGE₂ in the RORα expression. Finally, a significant RORα downregulation was observed in T cells permanently transfected with inducible MT₁ antisense. In conclusion, we show a novel connection between melatonin membrane receptor signalling and RORα expression, opening a new way to understand melatonin regulation in lymphocyte physiology. Received 23 September 2008; received after revision 19 November 2008; accepted 21 November 2008     

Oxygen consumption in rat skeletal muscle at various rates of oxygen delivery

Summary Resting oxygen consumption of the vascularly isolated gracilis anticus muscle of the rat perfused by natural circulation via the femoral artery with diluted or undiluted blood depends on oxygen delivery (the product of flow rate and arterial oxygen concentration-DO₂) only when DO₂ falls below 16 l/min/g.     

The plasmin–antiplasmin system: structural and functional aspects

The plasmin–antiplasmin system plays a key role in blood coagulation and fibrinolysis. Plasmin and α₂-antiplasmin are primarily responsible for a controlled and regulated dissolution of the fibrin polymers into soluble fragments. However, besides plasmin(ogen) and α₂-antiplasmin the system contains a series of specific activators and inhibitors. The main physiological activators of plasminogen are tissue-type plasminogen activator, which is mainly involved in the dissolution of the fibrin polymers by plasmin, and urokinase-type plasminogen activator, which is primarily responsible for the generation of plasmin activity in the intercellular space. Both activators are multidomain serine proteases. Besides the main physiological inhibitor α₂-antiplasmin, the plasmin–antiplasmin system is also regulated by the general protease inhibitor α₂-macroglobulin, a member of the protease inhibitor I39 family. The activity of the plasminogen activators is primarily regulated by the plasminogen activator inhibitors 1 and 2, members of the serine protease inhibitor superfamily.     

Differential vascular effects of neuropeptide Y(NPY) selective receptor agonists

Neuropeptide Y (NPY) increases blood pressure either directly or indirectly by potentiating the effect of various vasoconstrictors. Only one (the Y₁-receptor) of two subtypes of receptors (Y₁ and Y₂) is thought to mediate the vascular smooth muscle contraction. To test this hypothesis we challenged isolated rat mesenteric arteries that had a functional endothelium with (1–36) NPY and with specific Y₁-receptor ([Leu³¹, Pro³⁴] NPY) and Y₂-receptor ([Ahx^5–24, -Glu²--Lys³⁰] NPY) agonists. The Y₁-receptor agonist elicited a contractile response similar to that of NPY, whereas the Y₂-receptor agonist had no effect on wall tension. We also found that the presence of a functional endothelium has no influence on the contractile response to NPY. From these data we conclude that the direct contractile effect of NPY in the mesenteric artery is mediated by stimulation of Y₁-receptors and is not endothelium-dependent.     

Effect of hydrogen peroxide on the binding of Merocyanine 540 to human erythrocytes

The fluorescent dye Merocyanine 540 (MC540) is often used as a probe to monitor the molecular packing of phospholipids in the outer leaflet of biomembranes. In a previous study we showed that the increased staining of erythrocytes with a perturbed membrane structure was mainly due to an increase in the fluorescence yield of cell-bound MC540, rather than to an increase of the number of bound molecules. Erythrocytes and ghosts exposed to continuous fluxes of H₂O₂ exhibited pronounced lipid peroxidation. Further, red blood cells subjected to this form of oxidative stress also showed increased staining with MC540. It appeared that this was caused by a strong increase in binding of MC540, together with a slight red shift of the fluorescence emission maximum and a small increase in the fluorescence yield of bound MC540. The changed MC540 binding characteristics were not observed when lipid peroxidation was suppressed by the presence of the antioxidant BHT in the incubation medium. However, open ghosts exposed to H₂O₂ showed no increase of MC540 binding, excluding a direct involvement of lipid peroxidation. Measurement of fluorescence emission spectra and gel filtration studies showed that MC540 can bind to H₂O₂-exposed hemoglobin. Experiments with erythrocytes lysed in hypotonic medium after exposure to H₂O₂ revealed that peroxidation of lipids with H₂O₂ induced a non-specific permeabilization of the plasma membrane to MC540, thereby allowing MC540 to bind to the oxidatively denatured, more hydrophobic hemoglobin. These results indicate that conclusions about packing of phospholipids in the outer leaflet of the membrane based on increased MC540-staining should be drawn with care. Received 27 September 1996; received after revision 5 November 1996; accepted 27 November 1996     

Sur l'inversion de l'action hypertensive de la prostaglandine F2α, chez le rat

Summary In normal rats, prostaglandin F₂ is a potent vasoconstrictor. The corresponding systemic hypertension is progressively attenuated by tachyphylaxis, revealing a longlasting fall of blood pressure. Tachyphylaxis is accelerated by previous injection of arachidonic acid and bradykinin. The vasodepressive activity of PGF₂ is inhibited by indomethacin.

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Nous remercions vivement le docteurPike (Upjohn, Kalamazoo) qui nous a fourni gracieusement ces prostaglandines.     

Substance P-induced fibrinolysis in the forearm of healthy humans

Physiological saline with or without substance P (50 ng/ml) was infused into the humeral artery in 6 healthy males. Indices of fibrinolytic activity (whole blood diluted lysis time, euglobulin lysis time, lysis areas in non-heated fibrin plates produced by plasma or euglobulin precipitate, plasminogen plasma levels, ₂-macroglobulin, Cl-inhibitor, and ₂-antiplasmin) were evaluated in the homolateral antecubital vein before and after 5 min of substance P or saline infusion. After substance P the fibrinolytic activity increased, as can be seen from the shortening of lysis times (p<0.01) and enlargement of the lysis areas (p<0.01). A reduction of plasminogen plasma levels (p<0.01), associated with a decrease in ₂-anitplasmin (p<0.01), was also found. Alpha₂-macroglobulin and Cl-inhibitor were instead unaltered by the peptide. The saline infusion, on the other hand, was unable to modify any of the examined indices. We concluded that exogenous substance P given intra-arterially increases fibrinolytic activity in locally-sampled venous blood through a mechanism which remains to be elucidated.     

Extended blood half-life of monomethoxypolyethylene glycol-conjugated hen lysozyme is a key parameter controlling immunological tolerogenicity

The blood half-life of a protein is prolonged by conjugating a protein with a linear amphiphilic polymer, monomethoxypolyethylene glycol (mPEG). The conjugation gives a protein immunotolerogenicity; hence, it is likely that the long half-life is crucial for the tolerogenicity. We prepared a tolerogenic mPEG conjugate of hen egg lysozyme (mPEG_1.5-HEL), which is conjugated 1.5-fold the molecular weight of mPEG against that of HEL, and evaluated the relationship between in vivo stability and the tolerogenicity. mPEG_1.5-HEL retained immunogenicity to prime HEL-specific T cell and antibody responses and had a long blood half-life, more than 27 times that of native HEL. The tolerant state was maintained as long as mPEG_1.5-HEL was detected in sera. With a decrease in the blood mPEG_1.5-HEL level, the tolerant state returned gradually to the responsive state; however, reinjection of mPEG_1.5-HEL again restored the tolerance. Thus, the extended blood half-life of HEL by mPEG conjugation is probably vital for establishing and maintaining the tolerant states. Received 17 May 1999; accepted 1 June 1999     

Über eine neue Methode der Messung sehr kleiner Mengen von CO in Luft,Blut und Lunge

Summary We have developped a new method to detect and measure very small amounts of CO having a sensitivity of 1 p. CO per 100 million p. air. HgO is heated in an oven reaction chambre where it reacts with CO in releasing proportional amounts of mercury vapor; the latter is then measured quantitatively in an UV electronic spectrophotometer. Reaction chamber and UV meter are assembled into a handy portable unit. The CO amount in blood is measured by decomposing a sample of blood in a volumetric flask with H₂SO₄ and drawing the gas mixture through the unit. The CO amount in alveolar air is measured by means of breathing bags. It is thus possible to study the influence of subtoxic doses of CO and the cancerogenic action of tobacco smoking.     

Antidiuretic and thermogenic effects of intracerebroventricular prostaglandin H2 in ethanol-anaesthetized rats

Summary When PGH₂ was administered intracerebroventricularly at doses of 5 and 15 nmoles in ethanol-anaesthetized rats, alcohol diuresis was inhibited and rectal temperature, blood pressure and heart rate were all significantly increased.     

Association between pregnancy-associatedα 2-glycoprotein (α 2-PAG) and mixed leucocyte reaction determinants on the leucocyte surface

Summary ₂-PAG is present on the surface of mononuclear blood leucocytes and can be demonstrated predominantly on B-lymphocytes and monocytes. Pretreatment of cells with antibody to ₂-PAG leads to a marked reduction in Fc-rosette formation. Competitive blocking experiments with specific antisera reveal a particularly close asociation between ₂-PAG and MLR (mixed leucocyte reaction) determinants on the cell surface. These findings suggest one mechanism whereby ₂-PAG may modify cell-mediated immune responses.     

Plasma glycoproteins of diabetic and normal Chinese hamsters

Summary Plasma hexosamine levels were elevated in lines of diabetic Chinese hamsters when compared to nondiabetics. Plasma protein bound hexoses,a ₂ macroglobulin and total lipoprotein levels were not significantly correlated with either animal line or levels of fasting blood glucose.Acknowledgments. For the support which made this project possible, we wish to thank the Michigan Heart Association and The Upjohn Co.