Biomedical subjects

T M Price

Publications and source records attributed to T M Price.

At least 19 recordsLinked to original sources

PubMed1992

Effect of type 1 transforming growth factor-beta on the level of aromatase cytochrome P-450 in human fetal hepatocytes.

Aromatase cytochrome P-450 (P-450AROM) is the enzyme in the steroidogenic pathway controlling the formation of oestrogens from 19 carbon steroid precursors. Aromatase is present in various tissues of the human fetus. The liver is second only to the placenta in the level of P-450AROM activity in the fetus. In this study we examined the effects of type 1 transforming growth factor-beta (TGF beta) on P-450AROM expression in human fetal (HF) hepatocytes. The HF hepatocytes were dispersed into single cells which were placed into monolayer cell culture until confluent. Cells were then rinsed and treated in serum-free media with dibutyryl cyclic AMP (Bu2cAMP) for 72 h. Treatment with Bu2cAMP (2 mmol/l) caused a fivefold increase in aromatase activity in hepatocytes. The increase in aromatase activity apparently represented an increase in P-450AROM enzyme as determined by immunoblotting using an antibody directed against human placental aromatase. TGF beta blocked basal, as well as Bu2cAMP increases, in aromatase activity by over 50%. The effect of TGF beta was dose-dependent with maximal inhibition observed using 2-5 ng TGF beta/ml. Immunodetectable P-450AROM decreased in parallel with activity following TGF beta treatment. The mechanism of TGF beta action was not through increasing phosphodiesterase (PDE) breakdown of cAMP since inhibition of PDE had no effect on TGF beta action. Finally we examined the level of P-450AROM mRNA using competitive polymerase chain reaction amplification. Bu2cAMP increased mRNA levels of P-450AROM by 2.5-fold, while TGF beta inhibited this induction by 35%. The results of this investigation demonstrated that TGF beta is a potent regulator of P-450AROM expression in HF hepatocytes.

Aromatase↗

PubMed1990

Confluent durable endothelialization of endarterectomized baboon aorta by early attachment of cultured endothelial cells.

Since surgical endarterectomy produces a highly thrombogenic vascular flow surface, we evaluated in baboons the capacity of acutely attached, unspread, cultured, homologous aortic endothelial cells to form durable, confluent endothelial cell monolayers and to interrupt acute thrombus formation during exposure to arterial blood flow conditions. When cultured baboon aortic endothelial cells were incubated with segments of freshly endarterectomized baboon aorta for 20 minutes in vitro, 3.8 x 10(5) endothelial cells/cm2 became attached. The endarterectomized surface treated with endothelial cells showed the presence of rounded, unspread endothelial cells with intervening bare areas of vascular media. Platelet deposition onto the endothelial cell--treated surface during exposure to blood under high flow conditions for 1 hour in vivo was markedly reduced by the attached but unspread endothelial cells. Moreover, these attached endothelial cells underwent spreading on the endarterectomized surface in the presence of high shear blood flow to form a confluent endothelial monolayer at the end of 1 hour. Thus acutely attached cultured endothelial cells form a durable thromboresistant monolayer on surgically denuded vascular surfaces during exposure to arterial blood flow.

Animals↗

PubMed1989

Immunohistochemical localization of prostaglandin endoperoxide synthase in human fetal membranes and decidua.

Prostaglandins play an important role during the maintenance of pregnancy and the initiation of parturition. Prostaglandin endoperoxide synthase activity has been demonstrated in human fetal membranes and decidua. Using immunohistochemical techniques, we identified in these tissues the cell types that contain prostaglandin endoperoxide synthase. A total of 33 specimens, ranging from 8 wk to 42 wk gestation, were studied. Decidualized stromal cells stained the most intensely and consistently of all cell types. Cytotrophoblast of the chorion and early placental villi and syncytotrophoblast of all gestational ages demonstrated a lighter, more variable staining pattern. Regardless of gestational age, amnion stained in a heterogeneous fashion, with some cells demonstrating an intense staining and other cells having no staining. There were no observable differences in laboring compared to nonlaboring term specimens. In summary, the specific cell types that contain immunoreactive prostaglandin endoperoxide synthase have been identified in fetal membranes and decidua.

Blotting, Western↗

PubMed1988

Human immunodeficiency virus infection of T cells and monocytes proceeds via receptor-mediated endocytosis.

The rates of internalization and uncoating of 32P-labelled human immunodeficiency virus (HIV) in the human T lymphoid cell line CEM are consonant with a receptor-mediated endocytosis mechanism of entry. This interpretation was affirmed by electron microscopic observation of virions within endosomes. Virus binding and infectivity were inhibited to the same extent by pretreatment with OKT4A antibody, therefore, the CD4 receptor-dependent pathway of internalization appears to be the infectious route of entry. The pattern of internalization by the human monoblastoid cell line U937 proved to be more complex, involving rapid and efficient CD4-independent internalization. Electron microscopy revealed the presence of large intracellular vesicles, each containing several virions. Antibody against the CD4 receptor for virus efficiently blocked infection, but did not reduce significantly HIV binding or internalization in the U937 cell line. Consequently, U937 cells have a CD4-independent pathway of virus internalization that does not coincide with the route of entry for infectious HIV.

Antigens, Differentiation, T-Lymphocyte↗

PubMed1988

Durability of confluent endothelial cell monolayers on small-caliber vascular prostheses in vitro.

Since implantation of small-caliber vascular grafts with preformed confluent endothelial cell monolayers (ECMs) may prevent acute platelet deposition and thrombus formation, we have evaluated the conditions necessary to produce durable ECMs. Cultured human umbilical-vein ECs in buffer were attached to vascular grafts--either expanded polytetrafluoroethylene (ePTFE) or knitted Dacron, both with inside diameters of 4 mm--precoated with collaten type I and perfused in vitro with serum-containing culture medium to achieve cell spreading into confluent monolayers. The number of cells attached was quantified by DNA measurements and indium-111 labeling. Morphology was evaluated by scanning electron microscopy. The maximum number of cells that attached acutely was 3.6 X 10(5) cells/cm2 graft, and the minimum number needed for confluence was 1.4 X 10(5) cells/cm2 graft. Confluence was morphologically complete after 2 hours of in vitro perfusion at 15 ml/min. When ECMs were exposed to varying flow rates, cell retention after 1 hour was 96.1% +/- 5.6% at 50 ml/min, 94.6% +/- 6.1% at 100 ml/min, 77.6% +/- 10.8% at 200 ml/min (p less than 0.001), and 40.1% +/- 10.4% at 400 ml/min (p less than 0.0001). Confluence was maintained for all grafts exposed to flows of 100 ml/min or less. Fewer cells were retained when acutely attached, unspread cells (71.5% +/- 11.5%) were compared with established ECMs (89.9% +/- 6.7%) at a flow rate of 100 ml/min (p less than 0.0001). ECMs on knitted Dacron were more durable than on ePTFE (82.7% +/- 5.3% versus 75.5% +/- 4.8% remained attached at 200 ml/min, P less than 0.05). 111Indium-labeled and unlabeled cells were equivalent with respect to saturation level attachment, spreading time to confluence, and durability under flow at 100 ml/min. We conclude that confluent and durable endothelial cell monolayers can be established on small-caliber vascular grafts within 2 hours.

Blood Vessel Prosthesis↗

PubMed1988

Preformed confluent endothelial cell monolayers prevent early platelet deposition on vascular prostheses in baboons.

We assessed the capacity of preformed confluent endothelial cell (EC) monolayers on small-caliber prosthetic grafts to prevent early platelet deposition in a baboon model. Cultured human umbilical vein ECs were attached to expanded polytetrafluoroethylene (Gore-Tex, 4 mm inner diameter, 3 cm length) precoated with type I collagen and perfused in vitro for 2 hours at 15 ml/min with serum-containing culture medium to achieve cell spreading into confluent monolayers. Cell numbers were quantified by deoxyribonucleic acid assay or isotopic counting of indium 111-labeled ECs. Saturation density for cell attachment was 3.55 +/- 0.29 x 10(5) cells per square centimeter of graft. After 1 hour of in vitro perfusion at 100 ml/min, 92.8% +/- 1.8% of cells remained attached and the flow surface was morphologically confluent. When grafts were inserted as extension segments into arteriovenous silicone rubber (Silastic) shunts in baboons, thereby exposing the endothelialized grafts to native flowing blood (100 ml/min) for 1 hour, the EC monolayers remained confluent with 81.05% +/- 5.88% of the cells attached. Indium 111-labeled platelet deposition onto grafts was quantified by dynamic scintillation camera imaging. Platelet deposition on 10 endothelialized grafts was markedly reduced (0.16 +/- 0.04 x 10(9) platelets per graft) compared with 10 untreated control grafts (1.84 +/- 0.59 x 10(9) platelets, p less than 0.02), eight grafts with early attached unspread ECs (2.38 +/- 0.66 x 10(9) platelets, p less than 0.005), and 11 grafts treated with collagen alone (5.93 +/- 0.72 x 10(9) platelets, p less than 0.002).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1987

Inhibition of in vitro nuclear transport by a lectin that binds to nuclear pores.

Selective transport of proteins is a major mechanism by which biochemical differences are maintained between the cytoplasm and nucleus. To begin to investigate the molecular mechanism of nuclear transport, we used an in vitro transport system composed of a Xenopus egg extract, rat liver nuclei, and a fluorescently labeled nuclear protein, nucleoplasmin. With this system, we screened for inhibitors of transport. We found that the lectin, wheat germ agglutinin (WGA), completely inhibits the nuclear transport of fluorescently labeled nucleoplasmin. No other lectin tested affected nuclear transport. The inhibition by WGA was not seen when N-acetylglucosamine was present and was reversible by subsequent addition of sugar. When rat liver nuclei that had been incubated with ferritin-labeled WGA were examined by electron microscopy, multiple molecules of WGA were found bound to the cytoplasmic face of each nuclear pore. Gel electrophoresis and nitrocellulose transfer identified one major and several minor nuclear protein bands as binding 125I-labeled WGA. The most abundant protein of these, a 63-65-kD glycoprotein, is a candidate for the inhibitory site of action of WGA on nuclear protein transport. WGA is the first identified inhibitor of nuclear protein transport and interacts directly with the nuclear pore.

Acetylglucosamine↗

PubMed1987

In vitro endothelialization of small-caliber vascular grafts.

To establish the conditions for achieving immediate and complete endothelial cell coverage of the luminal surfaces of small-caliber (internal diameter:4 mm) vascular grafts in vitro, the attachment and spread of endothelial cells cultured from human umbilical veins to expanded polytetrafluoroethylene (PTFE) and knitted Dacron grafts was studied. Cell number was quantified by fluorescent measurements of deoxyribonucleic acid. The completeness of cell coverage and cell junction formation were assessed by means of both scanning and transmission electron microscopy. Cell attachment was compared after expanded PTFE or knitted Dacron grafts were precoated with gelatin, laminin, fibronectin, fibrin, or collagen, singly or in combination. Saturation cell attachment of 3.5 +/- 0.7 X 10(5) cm-2 was completed within 15 minutes when (1) type I collagen was used to form the substrate matrix, (2) human umbilical vein endothelial cells were suspended in phosphate-buffered saline solution supplemented with calcium and magnesium, and (3) the suspended cell number was greater than or equal to 5 X 10(5). In contrast, attachment to untreated or laminin-treated surfaces was 1.3 +/- 0.33 X 10(5) cells cm-2 and attachment to fibrin- or fibronectin-treated surfaces was 2.8 +/- 0.47 and 2.4 +/- 1.1 cells X 10(5) cm-2, respectively. However, to produce a confluent flow surface, the attached cells required several hours in culture medium to spread completely. Maintenance of confluent cell coverage on the graft surface for 3 days in vitro was achieved by means of continuous perfusion with oxygenated tissue culture medium RPMI-1640-HEPES supplemented with 20% fetal bovine serum. We conclude that optimum immediate confluent endothelial coverage of small-caliber vascular grafts requires a high concentration of cells, attachment to collagen-precoated grafts, and several hours of incubation in complete culture medium.

Blood Vessel Prosthesis↗

PubMed1985

Renal failure and hemolytic anemia associated with mitomycin C. A case report.

The authors report a case of renal failure and microangiopathic hemolytic anemia associated with the administration of mitomycin C in a 62-year-old man. Examination of the renal biopsy specimen revealed glomerular sclerosis and necrosis with interstitial fibrosis on light microscopy. Electron microscopic findings revealed deposits of fibrin within the glomeruli and interstitial tissue and separation of glomerular epithelial cells from the underlying lamina densa. Following plasma exchange, the hemolytic process improved, but renal failure persisted.

Anemia, Hemolytic↗

PubMed1985

The initial stages of adsorption of plasma derived proteins on artificial surfaces in a controlled flow environment.

Surfaces of amorphous carbon, polystyrene, and polycarbonate were exposed to solutions of fibrinogen, a modified fibrinogen which lacked the alpha chain protuberance, and serum albumin. The results were studied by electron microscopy. The exposures occurred in a well characterized flow environment at a shear rate of 135/s. All three protein species formed a film when adsorbed to carbon films. When the proteins were adsorbed to polystyrene surfaces, formation of a network was observed. Polycarbonate surfaces adsorbed the proteins both as a network and as a continuous film. It was observed that the nature of the adsorption process depended upon the specific combination of molecule and material. For example, on carbon, individual fibrinogen molecules retain their trinodular structure and adsorb randomly until a monolayer forms. On polystyrene, the individual fibrinogen molecules appear as globules and a network forms before complete coverage occurs.

Adsorption↗

PubMed1985

Amniotic fluid embolism. Three case reports with a review of the literature.

Amniotic fluid embolism is a catastrophic event of the intra- and early postpartum period which may also be seen with cesarean delivery and during abortions. Presenting symptomatology includes respiratory distress with cyanosis, shock, and possibly tonic-clonic seizures. DIC frequently occurs. The pathogenesis may include entry of amniotic fluid through lacerations or ruptures of the uterus or cervix, through endocervical veins and through abnormal uteroplacental sites, such as with placental abruption, placenta previa, or placenta accreta. Amniotic fluid probably causes cardiovascular-respiratory symptoms by pulmonary vascular obstruction and through a vasoactive substance causing pulmonary vascular constriction. The lethality of amniotic fluid may be enhanced by a high particulate content or meconium staining. The diagnosis of amniotic fluid embolism may be made ante mortem by demonstrating amniotic fluid debris in central blood samples or expectorated sputum. Postmortem diagnosis often requires meticulous examination of the pulmonary microvasculature with the utilization of special stains. Treatment is directed towards symptoms of shock, arterial hypoxemia, and DIC. Acute renal failure may complicate the picture after shock. If the patient survives the embolic and coagulative problems, recovery is usually complete without long-term sequelae.

Abortion, Missed↗

PubMed1982