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Biomedical subjects

M V Tahamont

Publications and source records attributed to M V Tahamont.

At least 19 recordsLinked to original sources
PubMed1991

Effect of fluid resuscitation from endotoxin shock on lung transvascular fluid and protein exchange.

The hypothesis that volume expansion during septic shock produces a greater transvascular protein flux than volume expansion alone was tested in anesthetized sheep by giving a high dose of endotoxin (40 micrograms/kg) intravenously. After 0.5 h of systemic hypotension, Ringer lactate, equivalent to 8% body wt, was infused followed by an additional 4 h of lymph collection. The results were compared with those from control animals receiving only Ringer lactate. The changes in plasma total protein with time were similar between groups. The increases in lymph flow and lymph protein flux were greater in the endotoxin-challenged group compared with the control group receiving Ringer lactate during the first 2 h but were similar thereafter. The interstitial volume was greater in the endotoxin-challenged animals compared with controls. The extravascular masses or apparent tissue concentrations for albumin or immunoglobulin G did not change in either group receiving Ringer lactate. The pulmonary edema following resuscitation from septic shock with Ringer lactate could be accounted for by either the pulmonary hypertensive effects of endotoxin or an initial, transient increase in microvascular protein permeability but not a sustained increase in microvascular permeability.

Animals↗
PubMed1989

Zymosan-activated plasma causes prolonged decreases in PMN superoxide release in sheep.

It is well established that activation of neutrophils within the pulmonary circulation produces acute lung injury in which adherence of neutrophils to endothelial cells is an obligatory step in the mechanism of injury. The effects of in vivo activation of neutrophils on the in vitro responses of these cells to stimulation have not been determined, although such information may be important in understanding how different etiological factors may interact to produce infection or acute respiratory failure. By using an assay to sequentially measure superoxide anion (O2-) release from adherent neutrophils stimulated with phorbol myristate acetate (PMA), we measured the in vitro activation response of peripheral blood neutrophils isolated before and 24 h after infusion of zymosan-activated plasma (ZAP; or untreated plasma as a control), air bubbles, or PMA in awake, instrumented sheep. Each of the three inflammatory agents produced an increase in lung microvascular permeability characteristic of acute lung injury; control plasma did not. For the in vivo ZAP experiments, stimulated O2- release in vitro by using PMA was approximately 50% lower (P less than 0.05) for neutrophils isolated 24 h after the in vivo infusion (4.3 +/- 0.8 nmol/500,000 cells) than before (8.1 +/- 0.2 nmol/500,000 cells). For the air emboli or PMA in vivo experiments, there were no changes in neutrophil activation responses in vitro. Similarly, infusion of control plasma did not result in reduced neutrophil O2- release. These results show that alterations in the inflammatory potential of neutrophils may occur in vivo and that such alterations appear to be dependent on the mechanism and agent by which lung injury is produced.

Animals↗
PubMed1988

Biochemical analysis of the activation of adherent neutrophils in vitro.

The role played by neutrophil oxidative responses in host defense and injury is an area of active investigation. In order to study neutrophil responses in vitro, methods are required for cell purification, enumeration, and quantification of activation responses, which mimic the in vivo situation as closely as possible. In this communication (and its companion paper, Albertine et al., 1988) improved methods for all of these tasks are described and applied to investigate neutrophil structure-function relationships in vitro and in vivo. Human neutrophils were purified by using a series of platelet-poor plasma-Percoll gradients (51, 62, 76 and 80% in Percoll). This modification of previously published procedures results in consistently successful neutrophil purification and has allowed us to purify neutrophils from bronchoalveolar lavage fluid as well as blood. Activation of human and sheep neutrophils (superoxide anion production) was quantitated by the reduction of ferricytochrome c using a microtiter plate reader to measure the increase in absorbance at 550 nm from adherent neutrophils. Adherence of neutrophils was quantitated by measurement of LDH in cells lysed with Triton X-100 using a new method which uses readily available commercial reagents and can quantitate the LDH content of as few as 5000 neutrophils (or the LDH released from 5% of 100,000 neutrophils). Assay conditions for superoxide anion were optimized, limitations both in assay design and instruments used to measure OD were explored and enumerated, and these methods were used to quantitate sheep and human neutrophil activation responses. Using methods described in Albertine et al. (1988) for fixing neutrophils in microtiter wells after assay of their functional capacity, we have studied the same cells functionally and morphologically. We have used these techniques to study blood and alveolar neutrophils from a patient with acute respiratory failure. His alveolar neutrophils displayed 67% of the activation response as peripheral neutrophils (4.31 +/- 0.12 nmol superoxide released per 250,000 neutrophils at 60 min vs. 6.38 +/- 0.18 in blood, P less than 0.01) and structural changes which suggested previous activation in vivo. These studies demonstrate that similar morphological changes are observed in neutrophils activated with phorbol myristate acetate in vitro, as are observed in cells which have been activated by pathophysiologic processes in vivo.

Animals↗
PubMed1988

Morphological analysis of the activation of adherent neutrophils in vitro.

One approach to study the inflammatory potential of neutrophils involves in vitro methods, using either adherent or suspended cells. In order to do structure-function studies, traditional methods require that experiments be done in parallel: one experiment for structure and another for function. In this report new morphological methods for coordinated structure-function studies on the same cells are described. Isolation and biochemical analysis of sheep and human adherent neutrophils were done as described in the companion paper (Cerasoli et al., 1988). Then, at designated time points, the adherent cells were fixed and processed in the microtiter wells for high-resolution light microscopy and transmission electron microscopy. The principal obstacle to the morphological studies was chemical etching of the microtiter wells by the processing solutions and embedding media. Insertion of No. 3 BEEM capsule 'sleeves' (the cap and conical tip were removed) into the wells before processing eliminated the obstacle and provided standard-sized, polymerized blocks for microtomes. Adherent neutrophils activated in vitro with 10(-7) M phorbol myristate acetate developed prominent cytoplasmic vacuoles. Furthermore, the activated cells assumed irregular shapes and cytoplasmic processes. These changes in adherent cell morphology in vitro are similar to those seen in neutrophils which have been activated and fixed in vivo. Thus, the in vitro approach we devised retains the morphologic characteristics of cells in vivo and provides an efficient method to do integrated structure-function studies. Using these techniques, we have studied alveolar neutrophils obtained from a patient with acute respiratory failure. These cells contained conspicuous cytoplasmic vacuoles, few granules, and their border was ruffled. The same morphologic changes observed after activation of peripheral blood neutrophils with phorbol myristate acetate in vitro were seen in the alveolar neutrophils obtained from this patient. Therefore, these studies reveal that similar morphologic changes are seen in neutrophils stimulated in vitro as well as cells which have been activated pathophysiologically in vivo.

Animals↗
PubMed1987

Prostaglandin E1 prevents increased lung microvascular permeability during intravascular complement activation in sheep.

Prostaglandin E1 (PGE1) inhibits a variety of functions of activated neutrophils including respiratory burst, release of leukotriene B4, and adherence to endothelial cells. To determine if PGE, alters the pathophysiology of complement-induced lung vascular injury, experiments were conducted in anesthetized sheep with lung lymph fistulas given a 1-hour infusion of zymosan-activated plasma. PGE1 (30 ng/min/kg) or its saline vehicle was infused intravenously for 90 minutes beginning 30 minutes before the infusion of activated plasma. PGE1 had no effect on leukocyte count, the initial hypoxemia and thromboxane A2 release, or the development of acute pulmonary hypertension. However, PGE1 prevented steady-state increases in lung lymph flow that in vehicle-treated sheep signaled an increase in lung microvascular permeability. Furthermore, extraction of PGE1 by pulmonary endothelial cells was unaffected by the infusion of activated plasma. We propose that PGE1 prevented the increase in lung vascular permeability by inhibiting adherence of activated neutrophils to endothelial cells.

Alprostadil↗
PubMed1986

The effects of cyclooxygenase inhibition on chemiluminescence and aggregation in sheep neutrophils.

Antiinflammatory actions of cyclooxygenase inhibitors may be related to inhibition of the synthesis and release of prostaglandins and thromboxane or to nonspecific actions of particular drugs. The role of cyclooxygenase products of arachidonic acid was studied in two leukocyte functions, free radical release and aggregation, after complement activation. Dose response curves were constructed after treatment with meclofenamate or ibuprofen. To differentiate between effects on free radical release from complement activated neutrophils and scavenging free radicals, additional experiments were made with a cell free system to generate free radicals. Both drugs inhibited complement initiated neutrophil chemiluminescence in a dose dependent manner. Meclofenamate acted primarily as a scavenger while ibuprofen inhibited free radical release. Neither drug had any inhibitory effects on complement induced leukocyte aggregation.

Animals↗
PubMed1986

Thromboxane as a mediator of pulmonary dysfunction during intravascular complement activation in sheep.

Intravascular complement activation results in thromboxane (TxA2) production, pulmonary hypertension, hypoxemia, and increased lung vascular permeability. The purpose of this study was to determine the role of TxA2 as a mediator of these responses. Experiments were made in anesthetized sheep subjected to intravenous injections of zymosan-activated plasma (ZAP) every 30 min for 4 h. Sheep were pretreated with dazoxiben, a TxA2 synthetase inhibitor, or SK and F 88046, a TxA2 end-organ antagonist, and the results were compared with those from untreated sheep. Dazoxiben, but not SK and F 88046, inhibited TxA2 release. The hypertensive response averaged 74 +/- 3 cm H2O after each injection of ZAP in untreated sheep. Neither drug altered this response. Pao2 decreased an average of 20 +/- 1 mmHg in untreated sheep, 3 +/- 1 mmHg in dazoxiben-treated sheep, and 11 +/- 1 mmHg in SK and F 88046-treated sheep. Increases in lung lymph flow and lymph protein clearance were unaffected by treatment. TxA2 appears to be an important mediator of hypoxemia during intravascular complement activation.

6-Ketoprostaglandin F1 alpha↗
PubMed1985

Dialysis-induced hypoxemia: membrane dependent and membrane independent causes.

Hypoxemia during hemodialysis may result from several differing processes. We initially studied patients undergoing standard acetate hemodialysis. At 15 minutes of dialysis, leukopenia (primarily neutropenia), a decline of platelet count, and hypoxemia occurred, but without a significant change in mean minute ventilation. Complement activation (V/A ratios of C5a greater than 1.0) persisted throughout dialysis. Leukocyte count returned to baseline by one hour. To separate the effects of solute and/or gas fluxes from those of blood-membrane interaction we studied changes in Po2, WBC, C5a, TxB2, and PGI2 during a period of blood membrane interaction without dialysis, and during subsequent acetate dialysis. Patients were studied with both polyacrylonitrile (PAN) and cuprophan membranes containing different priming solutions during membrane contact alone. Despite leukopenia and complement activation, hypoxemia failed to occur during membrane contact alone. At 15 minutes of subsequent acetate dialysis, significant hypoxemia occurred with both membranes. However, the degree of hypoxemia was twice as great with a cuprophan membrane primed with acetate (18.6 +/- 3.3 mm Hg) compared with air or bicarbonate (9.1 +/- 1.4 and 7.0 +/- 2.0 mm Hg, respectively), or compared with PAN (8 +/- 2.8 mm Hg). Changes in thromboxane B2, PGI2, and C5a did not correlate with changes in Po2. We conclude that there are two major components to dialysis related hypoxemia. One is membrane independent, and may relate to the metabolic effects of acetate or to dialyzer CO2 loss. The remaining portion is membrane dependent, occurring with cuprophan, but not with PAN, and is conditioned by an acetate dependent interaction between blood and membrane.

Adolescent↗
PubMed1985

Arachidonate cyclooxygenase metabolites as mediators of complement-initiated lung injury.

The pulmonary effects of intravascular complement activation can be divided into those that are initial events of activation and those that depend on the duration of intravascular activation. Pulmonary leukostasis is both an initial event and one that persists as long as intravascular complement activation is ongoing. This event is independent of synthesis and release of cyclooxygenase products of arachidonic acid. Initial events include synthesis and release of thromboxane and development of pulmonary hypertension and hypoxemia. Cyclooxygenase products are mediators of these initial pathophysiological events in part or in total. Complement-initiated increases in lung microvascular permeability are dependent on the duration of intravascular complement activation. Activation must be either prolonged or repeated to produce significant microvascular injury. This process is unrelated to synthesis and release of cyclooxygenase products, but is related to release of toxic oxygen metabolites.

Arachidonic Acids↗
PubMed1984

Aggregation and thromboxane synthesis and release in isolated sheep neutrophils and lymphocytes in response to complement stimulation.

In an attempt to determine possible cellular sources of thromboxane synthesis and release during intravascular complement activation, we prepared pure, viable fractions of neutrophils and lymphocytes from sheep blood. Lymphocytes were also isolated from thoracic duct lymph. Blood neutrophils and lymphocytes release thromboxane and the cells aggregate when challenged with zymosan-activated plasma. Blood lymphocytes release thromboxane in greater quantity than do neutrophils with the complement stimulus. Lymph lymphocytes, when stimulated with zymosan-activated plasma, did not release thromboxane or aggregate. These data suggest that both blood neutrophils and lymphocytes are likely sources of complement-initiated thromboxane synthesis and release in sheep.

Animals↗
PubMed1983

Granulocytes mediate the increase in pulmonary vascular permeability after thrombin embolism.

We examined the effect of pulmonary embolization with microthrombi on the lung vascular permeability to proteins and the role of platelets and granulocytes as putative cellular factors in mediating the alterations in permeability. Anesthetized artificially ventilated sheep were prepared with lung lymph fistulas. Pulmonary embolization was induced using thrombin. Pulmonary vascular resistance (PVR) was increased approximately threefold from baseline. Pulmonary lymph flow (Qlym) increased 2 h after thrombin, but the lymph-to-plasma protein ratio (L/P) did not change significantly from base line. Raising the pulmonary capillary pressure (Pc) by inflating a left atrial balloon produced a large increase in Qlym but no change in L/P, indicating a permeability-increasing effect of thrombin. Reduction of platelet count with antiplatelet serum before thrombin also produced an increase in Qlym without a change in L/P. Raising Pc in this group resulted in changes comparable with those in the control group, i.e., increased Qlym without a change in L/P. In contrast to both control and platelet-depleted groups, reduction of the granulocyte count with hydroxyurea did not affect Qlym or L/P after thrombin. Raising Pc in this group increased Qlym but decreased L/P, indicating normal capillary sieving of proteins. Therefore embolization of pulmonary vessels with microthrombi increases pulmonary vascular permeability, and the increase is mediated by granulocytes.

Animals↗
PubMed1983

Thrombin-induced lung vascular injury. Roles of fibrinogen and fibrinolysis.

We examined the contributions of fibrin and the fibrinolytic mechanism in mediating the thrombin-induced lung vascular injury. Studies were made in sheep in which alterations in lung transvascular fluid and protein exchange were assessed using the lung lymph fistula preparation. Group I(n = 10) control sheep in which left atrial pressure was raised to increase pulmonary lymph flow (Qlym); Group II(n = 8) control sheep in which thrombin was infused intravenously (58.3 +/- 12.6 U/kg) to induce pulmonary thromboembolization; Group III (n = 6) sheep were defibrinogenerated using Ancrod (purified fraction of venom of Agkistrodon rhodostoma) prior to embolization with thrombin (124.5 +/- 2.5 U/kg); Group IV (n = 13) sheep were treated with tranexamic acid to inhibit fibrinolysis prior to embolization with thrombin (140.0 +/- 25.8 U/kg). In Group II, pulmonary thromboembolization increased Qlym without a change in the steady-state lymph-to-plasma protein concentration ratio (L/P). Raising left atrial pressure (increases Pla) postembolization to test for an increase in pulmonary vascular permeability to proteins further increased Qlym but did not alter L/P, indicating an increase in permeability. In Group III, the thrombin-induced increase in Qlym was blunted and the L/P decreased; increases Pla further increased Qlym but decreased L/P. In Group IV, Qlym also increased after thrombin-induced embolization and L/P decreased; increases Pla further increased Qlym but decreased L/P. The responses to increases Pla of Groups III and IV were similar to the response of Group I rather than to that of Group II, indicating that defibrinogenation and fibrinolytic inhibition prevented the increase in lung vascular permeability. The results indicate that circulating fibrinogen and activation of plasmin are necessary for development of lung vascular injury after pulmonary thromboembolization.

Animals↗
PubMed1982

Mechanisms of lung vascular injury after intravascular coagulation.

Pulmonary intravascular coagulation and the resultant microembolization increase lung vascular permeability to proteins. The increase in permeability is mediated by the activation of cellular and humoral factors after intravascular coagulation. In particular, intravascular coagulation results in sequestration and activation of leukocytes, which appears to be of primary importance in mediating the lung vascular injury. In addition, fibrin entrapment and the generation of fibrin-degradation products after fibrinolysis also contribute to the vascular injury. The classical inflammatory agents, such as prostaglandin, histamine, bradykinin, and serotonin, may modulate the degree of injury after pulmonary vascular thrombosis, but they do not appear to be the primary mediators of the injury. Platelet aggregation did not mediate the lung vascular injury after intravascular coagulation, but was responsible for the pulmonary gas-exchange impairment that occurs with pulmonary vascular thrombosis.

Animals↗
PubMed1982

Lung fluid balance after pulmonary embolization: effects of thrombin vs. fibrin aggregates.

We examined the relationship between the activation of fibrinolysis and the increase in lung vascular permeability after pulmonary microembolization (PM). Sheep were prepared with lung lymph fistulas to assess pulmonary transvascular fluid and protein dynamics. Studies were made in three groups: group I (n = 8) in which PM was induced by an iv infusion of thrombin (60 +/- 13 NIH U/kg); group II (n = 7) in which PM was induced by an iv infusion of 50-micrometers-diameter fibrin microaggregates (0.32 +/- 0.009 g/kg); and group III in which the left atrial pressure was increased by 10-15 Torr by inflation of a balloon catheter. Thrombin caused an increase in pulmonary lymph flow (Qlym) without a change in the lymph-to-plasma protein concentration ratio (L/P ratio) indicating an increase in the lung vascular permeability to proteins. Fibrin microaggregates also increased Qlym, but the increase was associated with a decrease in the L/P ratio. The results in the latter group were similar to those obtained after left atrial hypertension in normal sheep. The increase in permeability after PM induced with thrombin was associated with large increases in the plasma concentration of fibrin degradation products, as compared with PM induced by fibrin microaggregates. The process of intravascular coagulation with the resultant generation of fibrinolysis and fibrin degradation products may be required for the increase in lung vascular permeability to proteins after pulmonary microembolization.

Animals↗