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PubMed1986

The effect of biliary obstruction on a gram-negative bacteremic challenge: a preliminary report.

Alterations in the sequestration and destruction of bacteria were studied after 10 days of biliary obstruction. Intraperitoneal injection of radiolabeled Escherichia coli was used to study bacterial localization in rats 10 days after common duct ligation and transection or sham celiotomy. Animals were sacrificed 4 hours later and uptake by liver, spleen, lung, and kidney were studied with a scintillation counter. No significant difference in localization between the two groups was noted. Bacteremia was induced in a second set of animals and quantitative bacterial organ cultures were performed. Significantly more viable organisms were identified in lung, liver, and kidney of animals that underwent common duct ligation and transection, when compared with controls that underwent sham celiotomy. This suggests that there is a defect in bacterial killing after 10 days of biliary obstruction. The inability to effectively clear and kill gram-negative bacteria in patients with biliary obstruction may account for some of the clinical complications seen in this patient population.

Animals↗

PubMed1993

Quantification of specific mRNA by flatbed scintillation counting of dual-labeled dot blots.

A dual-labeling technique was developed for direct quantification of specific mRNA using a flatbed liquid scintillation counter. This method simultaneously measures cpm of 32P- and 35S-labeled probes bound to RNA dot blots and subtracts counts due to nonspecific background radioactivity bound to the filter. Probes for T-cell receptor and beta-actin (as the internal standard) were hybridized both separately and simultaneously to RNA isolated from five different sources. There was concordance between the radioactivity measured from single- and dual-hybridizations for each combination of 35S- and 32P-labeled probes. This methodology directly quantifies specific mRNA sequences bound to membranes and has potential for measuring gene dosage, without the need for re-probing or densitometric analysis.

Actins↗

PubMed1987

Micromethod for assaying reverse transcriptase of human T-cell lymphotropic virus type III/lymphadenopathy-associated virus.

A micromethod for assaying the reverse transcriptase enzyme of human T-cell lymphotropic virus type III/lymphadenopathy-associated virus in cocultures of clinical specimens for viral isolation was developed and compared with the macromethod in use. Ultracentrifuged, pelleted, and solubilized viral culture supernatants were transferred into either tubes (macromethod) or microtiter plates (micromethod) and incubated with tritiated enzyme substrate. Trichloroacetic acid-precipitated DNA was collected on individual filter papers with a Millipore filtration manifold (macromethod) or on filter sheets using a semiautomated cell harvester (micromethod). Filters were then placed in scintillation fluid and counted on a beta scintillation counter. Results of the micromethod significantly correlated to those of the macromethod, with a linear relationship between the two. The cutoffs for positivity based on the mean + 2 standard deviations for a set of known negative specimens (n = 19) was 4,973 cpm for the micromethod compared with 5,336 for the macromethod. The intrarun and interrun variations were comparable for both methods. There was a 67% increase in the maximal daily number of specimens which could be run (100 versus 60) as well as a reduction in reagent use. In summary, the micromethod utilizing a semiautomated cell harvester is comparable to the existing macromethod in accuracy and is an improvement due to savings in time and reagents.

HIV↗

PubMed1992

A simple quantitative assay for chloramphenicol acetyltransferase by direct extraction of the labeled product into scintillation cocktail.

A simple, rapid, sensitive, quantitative, and inexpensive assay for chloramphenicol acetyltransferase (CAT) is described. The assay is based on the direct extraction of the products of the reaction into toluene-based liquid scintillation cocktail. The assay is carried out in 7-ml scintillation vials using 1 mM chloramphenicol and either 100 microM acetyl-CoA and 0.1 microCi of [3H]acetyl-CoA or 1 mM acetyl-CoA and 0.5 microCi of [3H]acetyl-CoA. After incubation, the reaction is terminated with 0.5 ml of 0.1 M sodium borate-5 M NaC, pH 9. The acetylchloramphenicols are extracted with 5 ml of 0.4% 2,5-diphenyloxazole-0.005% 1,4-bis(5-phenyloxazol-2-yl)benzene in toluene by a 30-s shaking. After a short centrifugation to clarify the layers, the vials are counted in a liquid scintillation counter. Extracted products are stable in the organic layer. Under these conditions, nearly 100% extraction of acetylchloramphenicols is shown using nonlabeled compounds and spectrophotometric methods. Using pure enzyme in the assay, linearity of activity with enzyme concentration, time, and temperature of incubation is demonstrated. Assays may even be carried out at 60 degrees C, where the enzyme activity is 3.4-fold higher than that at 23 degrees C. The increase in enzyme activity with increasing temperature is due to the increased formation of predominantly 3-acetyl and 1-acetylchloramphenicols and not to 1,3-diacetylchloramphenicol. The present assay compared very well with the standard assay using [14C]chloramphenicol and TLC. Using this assay, we measured quantitatively the CAT activity in extracts of pSV2-CAT-transfected CV-1 cells in 10 min and NIH 3T3 cell extracts in 60 min at 60 degrees C.(ABSTRACT TRUNCATED AT 250 WORDS)

3T3 Cells↗

PubMed1979

[Methodical optimization of redioiodine uptake experiments in living rats].

The paper deals with the estimation of thyroidal uptake of 131I in living rats. The animals are fixed in a specially marked glass tube. This tube is discontinuously moved over a scintillation counter within a lead collimator. Counts of 131I are estimated segmentically. The highest counts ratio with the geometrical factor of the appropriate segment is used to calculate the thyroidal radioiodine uptake. Similar results, obtained with an 131I-source placed in various segments of the top side of glass tube, indicate that the uptake values obtained in this manner exact. Thyroidal uptake values, which were received on living rats with this method (in vivo) and compared with values obtained with the prepared and plated thyroid of the same rats (in vitro) show a very high correlation (r = 0,99; p greater than 0,001). In repeated estimations of the thyroidal 131I-uptake on one animal a variation coefficient of 1.5% (n = 13) was obtained. The advantage of this in vivo method is the possibility to determine the thyroidal activity at various times after 131I-application (2 phase test) and by repeated 131I-applications under different conditions (diet, age, for instance).

Animals↗

PubMed1999

Radiolabeled thymidine: a sensitive tracer for early tumor response and recurrence after irradiation.

UNLABELLED: This study evaluated the sensitivity of a radiolabeled thymidine tracer for assessment of early tumor response and recurrence after irradiation. METHODS: SW707 human colon carcinoma implanted into nude mice was irradiated with 6 or 20 Gy. Tumor volume was determined for an interval of 14 d. At 4, 8 and 24 h and at 2, 3, 7, 10 and 14 d after irradiation, [14C]thymidine uptake into the tumor was determined with a liquid scintillation counter and the intratumoral distribution of [14C]thymidine was visualized and evaluated semiquantitatively by autoradiography using a phosphor imager. RESULTS: In both groups, tumor volume decreased until day 7 after irradiation; afterward, regrowth occurred in only the group that had received 6 Gy. A decrease in thymidine uptake was found as early as 8 h after irradiation. On day 3 after irradiation, thymidine uptake increased again in the 6-Gy group, before the increase in tumor volume, but remained unchanged in the 20-Gy group. Also on day 3, multiple foci of thymidine uptake suggesting proliferation preceding tumor recurrence were seen on autoradiographs from the 6-Gy group but not from the 20-Gy group. Histological findings correlated with the results of autoradiography. CONCLUSION: The results show that radiolabeled thymidine is a sensitive tracer for assessment of early tumor response and recurrence after irradiation. The rapid decrease in uptake, however, does not allow any prediction about tumor recurrence.

Animals↗

PubMed1978

A new assay for rapid measurement of MIF levels by 3H-labelled cells in liquid scintillation counting vials: statistical implications for the measurement of migration inhibition.

A new quantitative assay for migration inhibitory factor (MIF) employs 3H-labelled cultured mouse or human lymphoid cells migrating from capillary tubes. Capillaries filled with labelled cells are placed in liquid scintillation counting vials, along with the MIF-containing sample and are removed at the end of a five-hour incubation period. The residual, labelled cells which have migrated out of the tubes are solubilized and counted in a liquid scintillation counter. While cultured lymphoblast cells are routinely used in the assay, the method was checked against mouse and guinea pig peritoneal exudate cells in both the labelled cell technique and the conventional chamber assay. The assay is technically simple to perform and a useful tool for laboratory research purposes because of the short span of time needed to obtain the results. These advantages indicate a potential for automation and use of this assay in a clinical immunology laboratory. Statistical analysis of data from both assays demonstrated that the relative variation among replicates is lower in the labelled cell assay. The new assay also measured a significant difference between controls and MIF-containing samples when the migration index (MI) was greater than 80%. Criteria for significant inhibition of migration are discussed in regard to the use of analysis of variance (ANOVA) and other statistical procedures, and the inadequacy of a single measure, such as the MI, is discussed.

Animals↗

PubMed2001

Age-related differences in fat-free mass, skeletal muscle, body cell mass and fat mass between 18 and 94 years.

OBJECTIVE: To determine (1) lean and fat body compartments, reflected by fat-free mass (FFM), appendicular skeletal muscle mass (ASMM), body cell mass (BCM), total body potassium (TBK), fat mass and percentage fat mass, and their differences between age groups in healthy, physically active subjects from 18 to 94 y of age; and (2) if the rate of decrease in any one of the parameters by age might be accelerated compared to others. METHODS: A total of 433 healthy ambulatory Caucasians (253 men and 180 women) aged 18--94 y were measured by dual-energy X-ray absorptiometry (DXA) and whole body scintillation counter (TBK counter) using a large sodium iodide crystal (203 mm diameter). RESULTS: The ASMM change (-16.4 and -12.3% in men and women, respectively) in >75 y-old compared to 18 to 34-y-old subjects was greater than the FFM change (-11.8 and -9.7% in men and women, respectively) and this suggests that skeletal muscle mass decrease in older subjects was proportionally greater than non-skeletal muscle mass. BCM (-25.1 and -23.2% in men and women, respectively) and TBK differences were greater than the differences in FFM or ASMM suggesting altered composition of FFM in older subjects. Women had lower peak FFM, ASMM, BCM and TBK than men. CONCLUSIONS: The decline in FFM, ASMM, BCM and TBK is accelerated in men and women after 60 y of age and FFM, ASMM, BCM and TBK are significantly lower than in younger subjects. Fat mass continued to increase until around 75 y.

Absorptiometry, Photon↗

PubMedDate not supplied

99mTc-pyrophosphate kinetics in man (relationship between bone uptake and mineral content).

Both the kinetics and the regional distribution of 99mTc-PPi were studied in the skeleton of healthy subjects of various ages by surface counting using a scintillation counter placed over different skeletal areas. The results were compared with bone mineral content as determined by the photonic absorption technique. Kinetic data of bone uptake were obtained by subtracting the counts measured in blood from the counts recorded over each body surface after making proportional corrections. The surface-counting patterns show a higher generalized 99mTc-PPi bone uptake in children and adolescents than in adults. The relative regional distribution of the tracer shows no difference between the two groups. The tracer accumulated more rapidly in spongy bone than in compact bone, and the fixation process was carried out during the first 30 min for both spongy and compact bone. In youths, however, higher uptake values corresponded to lower values of mineral content. The findings enable the normal surface-counting pattern to be constructed and confirm that the 99mTc-PPi uptake is related to bone metabolism; they also suggest the application of external measurements for the study of metabolic bone diseases.

Adolescent↗

PubMed1999

Strongyloides ratti: migration study of third-stage larvae in rats by whole-body autoradiography after 35S-methionine labeling.

In order to clarify the migration pathway of Strongyloides ratti, Wistar rats were given 5,000 35S-labeled infective larvae subcutaneously and killed at 10, 15, 20, 25, 30, 40, and 50 hr postinfection. Prior to inoculation, the specific radioactivity level was assessed in the labeled larvae using a scintillation counter. The frozen rat specimens were sectioned at 50 microm, and the sections were freeze-dried and mounted on X-ray film in darkness. The labeled larvae appeared as dark spots on the film after 14 days of exposure. The infected larvae remained at the inoculated site (lower abdomen) until 10 hr after infection. Some larvae were found in the head portion, whereas others existed sporadically in the skin, liver, and lungs at 15 hr. After 20 and 25 hr, the majority of larvae had accumulated in the head portion. Many larvae appeared in the cranial and nasal cavities; however, no larvae were found in any other organs or tissues. At 30 hr, most larvae had begun to accumulate in the ethmoid region again. At 40 and 50 hr, some larvae were recognized in the ethmoid region, and most had already reached the small intestine. This suggests that the larvae directly move to the nasofrontal portion through the subcutis, rather than migrating to the head through either the viscera, ascending vessels, or the foramen occipital magnum.

Animals↗

PubMed2003

Comparison of 99mTc-MIBI uptakes on planar images with those in excised rats organs.

The precision with which images reflect tracer uptake in the myocardium has been studied. Additionally, the degree to which Tc methoxyisobutylisonitrile (99mTc-MIBI) in the liver gave the effect to a myocardial image has been examined. After administering Tc-MIBI to normal male rats, we compared the myocardial uptakes obtained using a gamma camera with the actual uptakes in the excised organs. Twenty-nine rats were used. Following imaging, the anterior view at 5, 10, 15, 30, 45, 60, 90 and 120 min after administration of the tracer, uptakes in the heart, lung, liver and blood were estimated with a well-type scintillation counter (WC) and represented as the percentage of the injected dose per gram of tissue (%ID/g). The regions of interest (ROIs) were placed on planar images (PI) and the uptake in each organ was estimated as the percentage of the injected dose per pixel (%ID/pixel). The ratios of PI-to-WC and heart-to-organ were also evaluated. Cardiac uptake with WC was maximum (1.581%+/-1.893%) at 10 min post-injection. On the other hand, that with PI was maximum (1.493%+/-0.598%) at 45 min post-injection, but there were significant differences between both measurements (PI/WC ratio: about 1.0 time). Pulmonary uptake with WC was the maximum at 5 min (0.808%+/-0.015%) post-injection, and decreased gradually. PI measurement showed the maximum value at 45 min (0.760%+/-0.012%). Hepatic uptake with WC was the maximum at 30 min (0.594%+/-0.254%). On the other hand, PI measurement showed the same pattern with WC, but these values were higher value than WC as the whole. PI measurement showed higher uptakes in each organ than WC measurement. It was concluded that uptakes or the heart-to-organ ratio obtained clinically with PI might not represent a value that is always accurate.

Animals↗

PubMed2003

Effects of disodium ascorbyl phytostanol phosphates (FM-VP4) on cholesterol accumulation within rat intestinal cells.

The objective of this study was to determine whether FM-VP4, a novel compound derived from plant sterols, can effectively reduce cholesterol accumulation within rat intestinal epithelial crypt (IEC-6) cells. EC-6 cells were cultured in Dulbecco's minimal essential medium (DMEM) containing 5% fetal bovine serum, 100 U/mL penicillin, 100 micro g/mL streptomycin, and 0.1 units/mL insulin at 37 degrees C under a humidified 5% CO2 atmosphere and seeded at 6.4 x 10(4) cells/well in 48-well plates. Experiments were initiated 14 days postconfluence. IEC-6 cells were exposed to [3H]cholesterol micelles (containing oleic and taurcholic acids), co-incubated with FM-VP4 (0, 10, 50, and 100 micro M) in Hepes Buffered Sterile Saline (HBSS). Cells were also preincubated with FM-VP4 prior to [3H]cholesterol micelle incubation to determine whether its effects are elicited intracellularly. The cellular localization of cholesterol was determined using digitonin. To determine the effects of cholesterol on the extent of FM-VP4 accumulation within IEC-6 cells, [3H]FM-VP4 was incubated with IEC-6 cells in the presence of unlabeled cholesterol micelles (0, 10, and 50 micro M). The extent of [3H]cholesterol or [3H]FM-VP4 associated with cell monolayers was determined after cell lysis using liquid scintillation counting in a Beckman LS6500 Scintillation Counter. Dose-response and time course studies were performed in which control (no FM-VP4 treatment) and FM-VP4 (10-100 micro M) were co-incubated with 50- micro M [3H]cholesterol micelles from 1 minute to 24 hours. Incubation with only 50- micro M FM-VP4 for less than 24 hours resulted in a 50% to 60% reduction (n = 6, P <.05) in [3H]cholesterol associated with the monolayer compared with control (n = 6). Preincubation of FM-VP4 did not elicit a significant reduction in cholesterol accumulation compared with control (n = 6). Approximately 25% of the total [3H]cholesterol associated with the cells was determined to be cytosolic, while 75% was noncytosolic in the presence and/or absence of FM-VP4. [3H]FM-VP4 was also shown to associate with IEC-6 cells at similar concentrations to cholesterol with the most pronounced inhibition of FM-VP4 accumulation occurring at a cholesterol concentration of 50 micro M. However, cholesterol-induced inhibition was detectable only after 1 hour of incubation. FM-VP4 inhibits cholesterol accumulation within IEC-6 cells and is most effective at equimolar concentrations with cholesterol. Our findings further suggest that the action of FM-VP4 is likely at the cell surface and not elicited intracellularly.

Animals↗

PubMed2001

Quantitative assay of telomerase activity in head and neck squamous cell carcinoma and other tissues.

OBJECTIVES: To confirm the applicability and use of a new technique to detect and quantify telomerase activity of specimens from head and neck malignant neoplasms and to explore whether the levels of telomerase activity can be a useful marker for cancer risk assessment in head and neck malignant neoplasms. DESIGN: Ninety-six specimens from 39 patients with head and neck malignant neoplasms were obtained. The specimens included 39 from patients with primary tumors (25 with head and neck squamous cell carcinoma and 14 with others), 10 from patients with neck metastases, 10 from patients with dysplasias, and 37 from patients with normal tissue. HeLa cell lines were used as positive control samples. MAIN OUTCOME MEASURE: The levels of telomerase activity were determined using a liquid scintillation counter. RESULTS: The new method has a high rate of outcome reproducibility. The intrabatch and extrabatch variations were 15.6% and 16.4%, respectively. The linear relationship was good between the telomerase activity and the value within 700 radioactive cpm (rcpm) to approximately 7000 rcpm. The levels of telomerase activity determined by radioactive count were more than 1000 rcpm in 42 of the 49 malignant specimens and much more than that in the normal tissues, with the exception of 3 specimens. The levels of telomerase activity in normal tissues were less than 1000 rcpm in every sample and less than that in the malignant neoplasm samples, with the exception of 1 specimen (P < .000). Higher levels of telomerase activity in 2 of 10 tissues from patients who had dysplasias were detected (2 specimens from patients who had severe dysplasia). The differences in the levels of telomerase activity between the head and neck squamous cell carcinomas and the other tumors were not statistically significant (P > .05). CONCLUSIONS: Detection of telomerase activity in head and neck malignant neoplasms can be a useful marker for the assessment of cancer. Telomerase reactivation may play an important role in tumorigenesis in head and neck squamous cell carcinoma. The quantification of telomerase activity may have clinical diagnostic value for head and neck malignant neoplasms.

Adult↗

PubMed1990

[Effect of heavy metal ions on the cells derived from human periodontal ligament. Effects of Pb and Cd].

It has been thought that the incorporation of Pb and Cd into human body, especially calcified tissues including the tooth, has altered the physiological processes of development. But, the biochemical events that initiate this process remain quite unknown. The present study attempt to explore the effect of Pb and Cd on human periodontal ligament fibroblast-like cells of the permanent tooth (HPLF) and the deciduous tooth (HPLF-Y) with respect to the cell growth, ALPase activity and incorporation of 14C-amino acids. HPLF and HPLF-Y migrated from a explant and subcultured according to the previously described method of Saito were inoculated 1.25 x 10(4) cells/cm2 in D-MEM supplemented with 2 mg/ml FCSP, 50 micrograms/ml ascorbic acid and antibiotics. After 24 hrs, HPLF and HPLF-Y were treated every two days for 10 days with 0-200 microM Pb or 0-10 microM Cd. Protein contents, DNA contents and ALPase activity were determined by Bio-Rad protein assay, deaminobenzoic acid assay and p-nitrophenylphosphate (pH 10.15) assay respectively. At 6 days HPLF were incubated with 3H-thymidine (TdR 2.0 microCi/well) and then the incorporation of 3H-TdR into cold TCA precipitates was assayed by a liquid scintillation counter. And also, at 6 days HPLF and HPLF-Y were incubated with 14C-amino acids (2.0 microCi/60 mm dish) for 24 hrs. The cell layers labeled with 14C were extracted with 15 mM Tris-HCl buffer containing 7 M urea (pH 7.4) and applied to the gel permeation chromatography of HPLC system to separate the fractions according to molecular weight. The HPLF and HPLF-Y incubated with Pb and Cd were morphologically identical. Pb stimulated the protein contents of extracellular matrix of HPLF, but not HPLF-Y. Cd inhibited the protein contents of cell layers of HPLF and HPLF-Y. With increasing concentrations of Pb and Cd, the incorporation of 3H-TdR into HPLF was inhibited. On the other hand, Cd stimulated the ALPase activity per DNA content since the ALPase activity of HPLF and HPLF-Y was decreased by the addition of Pb. The distribution of 14C-labeled protein according to molecular weight did not alter the chromatographic pattern of HPLF incubated with Pb and Cd. But, that of HPLF-Y incubated with Pb was relatively shifted to low molecular size. Therefore, these responser concluded that HPLF were not completely identical with HPLF-Y. Pb and Cd not only had a toxic effect on cell growth, but also they may regulate the metabolic alteration.

Alkaline Phosphatase↗

PubMed1999

Prediction of in vivo background in phoswich lung count spectra.

Phoswich scintillation counters are used to detect actinides deposited in the lungs. The resulting spectra, however, contain Compton background from the decay of 40K, which occurs naturally in the striated muscle tissue of the body. To determine the counts due to actinides in a lung count spectrum, the counts due to 40K scatter must first be subtracted out. The 40K background in the phoswich NaI(TI) spectrum was predicted from an energy region of interest called the monitor region (95-129 keV), which is above the 238Pu region (11-32 keV) and the 241Am region (40-82 keV), where photopeaks from 238Pu and 241Am occur. Empirical models were developed to predict the backgrounds in the 238Pu and 241Am regions by testing multiple linear and nonlinear regression models. The initial multiple regression models contain a monitor region variable as well as the variables gender, (weight/height)alpha, and interaction terms. Data were collected from 64 male and 63 female subjects with no internal exposure. For the 238Pu region, the only significant predictor was found to be the monitor region. For the 241Am region, the monitor region was found to have the greatest effect on prediction, while gender was significant only when weight/height was included in a model. Gender-specific models were thus developed. The empirical models for the 241Am region that contain weight/height were shown to have the best coefficients of determination (R2) and the lowest mean squares for error (MSE).

Actinoid Series Elements↗

PubMed1987

Use of in vivo/in vitro unscheduled DNA synthesis for identification of organ-specific carcinogens.

There are still only a few in vivo short-term assay methods for predicting potential organ-specific carcinogens and mutagens in mammals, although such methods are required for evaluating the in vivo effects of in vitro mutagens. In the in vivo/in vitro UDS assay methods described here, chemicals are given to experimental animals and induction of UDS in target organs is determined by in vitro organ culture or primary cell culture in the presence of [3H]dThd. Incorporation of [3H]dThd into DNA is measured with a liquid scintillation counter or by autoradiography. These methods have now been applied to the glandular stomach, forestomach, colon, liver, kidney, pancreas, tracheal epithelium, nasal epithelium, and spermatocytes. With minor modifications, they may also be applied to other organs. The present review shows that induction of UDS in various organs correlated well with the induction of cancer in these organs. The present authors have used the present methods to identify some potential organ-specific mutagens and carcinogens in mammals. The present authors found that three dicarbonyl compounds, glyoxal, methylglyoxal, and diacetyl, induced apparent UDS and TDS in the glandular stomach, and other groups found that 2-NT, MA6BT, and CNEt6BT induced UDS in the liver. These in vivo/in vitro UDS assays are better than in vitro UDS assay for identification of potential organ-specific mutagens and carcinogens in mammals and are especially useful for identifying potential mutagens and carcinogens that are specific for certain organs, such as the stomach, liver, and kidney. They are also useful for examining the potential mutagenicities and carcinogenicities of carcinogen analogs. However, these methods are not suitable for general in vivo screening because they are not yet available for all organs. A further advantage of the methods is that they can be used to examine larger numbers of animals at one time than other methods for detecting DNA damage, such as alkaline elution or alkaline sucrose density gradient centrifugation. Glyoxal enhanced cancer induction in the glandular stomach by the administration of a limited amount of MNNG and then glyoxal afterward in the two-stage stomach carcinogenesis.

Animals↗

PubMed1996

The effect of leukocytes on adhesion molecules. An explanation of blood transfusion enhancement of tumor growth.

OBJECTIVE: To determine the mechanism of the adverse relationship between perioperative blood transfusion and lung cancer recurrence, by focusing on endothelial cell adhesion molecules (CAMs), which are thought to play a role in distant tumor cell implantation. DESIGN AND OUTCOME MEASURES: Murine endothelial cells were cocultured with allogeneic leukocytes, syngeneic leukocytes, and syngeneic lung carcinoma cells for 60 hours. The percentage of endothelial cells expressing vascular CAMs (VCAMs) and intercellular CAMs (ICAMs) was quantified during this time using indirect immunofluorescence and flow cytometry. Tumor cell adhesion to the endothelium was quantified for 6 hours using cells labeled with sulfur 35 and a scintillation counter. SETTING: Laboratory. MATERIALS: C57/BL and Balb/C mice. RESULTS: Vascular CAM was not expressed on the endothelium, but ICAM was preferentially expressed without stimulation. Tumor-cell adhesion and endothelial ICAM expression were inversely related. After 15 hours of coculture, tumor cell adhesion was four-fold greater in the experimental group than in the control, and coincident endothelial ICAM expression was fourfold lower. CONCLUSION: Endothelial cell ICAM expression is negatively correlated with metastatic potential.

Animals↗

PubMed1999

Photobinding of [gamma-(32)P] ATP gamma-benzophenone to the surface of a polyurethane membrane in the preparation of a beta-particle-emitting balloon catheter.

PURPOSE: The goal of this study was to photochemically bind 5'-[gamma-(32)P]-azido-ATP gamma-benzophenone ((32)P-ATP-BPA) to a polyurethane surface. Expandable balloon catheters composed of (32)P-coated polyurethane have the potential for preventing restenosis following percutaneous transluminal coronary angioplasty. METHODS: After extensive preparation and cleaning of polyurethane disks, 10 microL of the radioactive ATP-BPA reagent (specific activity = 9.4 Ci/mmol) was applied to the surface. After drying, the membrane disks were exposed ultraviolet radiation (254 nm; 6,000 microwatts/cm(2)) for up to 2 h and subsequently washed. The amount of (32)P bound to the membrane disks was determined by Cerenkov counting in a liquid scintillation counter. The effect of the labeling solution composition (solvent, presence of potassium or manganese ions, addition of surfactants, etc.) on photobinding efficiency was determined. RESULTS: The efficiency of attaching the (32)P-ATP-BPA reagent to the polyurethane surfaces was markedly dependent upon the cleaning and pretreatment conditions. Following detailed washing and rinsing steps, a photobinding efficiency of 36.4+/-3.6% was obtained with 10 min UV exposure time using (32)P-ATP-BPA solutions that were 95/5 methanol/water by vol. Increasing the concentration of the (32)P-ATP-BPA reagent did not improve the photobinding efficiency; however, the total amount of (32)P bound to the disks was increased. CONCLUSIONS: Photochemical methods can be employed to attach beta(-)-emitting radionuclides to polymers that are employed as balloon catheters. The preparation of the polymeric material (washing, rinsing, and drying) is critically important in maximizing the amount of (32)P-ATP-BPA that can be bound to the polymer.

Adenosine Triphosphate↗