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Biochemical study of human periodontal ligament: preparation of cell attachment materials induced by pulsed electromagnetic fields.

The periodontium, especially the periodontal ligament and alveolar bone, are tissues constantly subjected to physical stress such as occlusion and mastication. This study was designed to explore the effect of the pulsed electromagnetic fields (PEMF) on the cell attachment and the spread of human periodontal ligament fibroblasts (HPLF) and rat osteoblasts (ROB). PEMF are categorized as one type of mechanical stress. HPLF were obtained by the explantation method described by Saito et al. They were then subcultured in Dulbecco's modified Eagle's medium (D-MEM) and supplemented with 2 mg/ml dialyzed fetal calf serum protein (FCSP), 50 micrograms/ml ascorbic acid and penicillin/streptomycin after trypsinization. ROB were isolated from a two-day-old rat calvaria by the sequential bacterial collagenase digestion method described by Dziak and Brand and were subcultured in D-MEM supplemented with FCSP, ascorbic acid and penicillin/streptomycin. After the confluent HPLF were cultured with serum-free MCDB 107 medium, the quiescent HPLF were exposed with or without PEMF for 24 hr. This was followed by the collection of the control conditioned medium (C-CM) and PEMF exposed conditioned medium (PEMF-CM). The cell attachment assay was performed so that the hydrophobic 24 multiwells were coated with the whole conditioned medium or fractionated conditioned medium by a PO-60K column. After coating, heat inactivated BSA blocked nonspecific sites for cell adhesion, and 3H-TdR labeled HPLF or ROB were cultured on the precoated wells. The activity of cell attachment and spreading was determined by the radioactivity of 3H-TdR using a scintillation counter. The characters of cell attachment factors derived from HPLF were hydrophobic, heat labile and proteolytic enzyme digestible. In addition, the fractionated PEMF-CM enhanced the spreading activity of ROB. PEMF induced the 10 KDa which can enhance the HPLF and ROB spreading. Therefore, the cell attachment and spreading factors secreted by HPLF exposed with PEMF may regulate HPLF and also ROB.

Animals↗

[Effects of EDTPO on hard tissues in rats. 2. Changes of serum components and morphology of hard tissues due to various doses of EDTPO].

Dose dependent effects of EDTPO on morphology and mineralization of hard tissues as well as calcium (Ca), inorganic phosphorus (IP) and alkaline phosphatase (ALP) levels in serum were examined in growing rats. Five groups of male S.D. rats weighing 110-120g were received s.c. injections of 0 (control), 1, 2, 4, and 8mg P/kg body wt. of EDTPO once daily (at noon) for 10 days respectively. Tetracycline (Tc.) were i.p. injected three times, one day before drug treatment (1st), 5th (2nd) and 10th (3rd) day of experimental period. All rats were received i.p. injection of 185kBq (5 microCi) of 45Ca six hours after the 10th injection of EDTPO. On the next day, blood samples were taken from the carotid artery. Then the animals were killed and both tibias, submandibular teeth and right femur were removed. Serum Ca, IP and ALP were determined by spectrophotometry. Wet, dry and ash bone weight were measured and bone calcium was determined by atomic absorption method. 45Ca activity was counted by the liquid scintillation counter. Serum Ca levels revealed decreasing tendency at the dose of 1mg P/kg and 2mg P/kg of EDTPO but increasing tendency at 4mg P/kg and significant increase at the dose of 8mg P/kg. Specific activity of 45Ca in serum increased slightly at 2mg P/kg, but at the dose of 8mg P/kg increased five times compared to that of control. Decrease of IP and increase of ALP level were observed only at the dose level of 8mg P/kg. Wet, dry, and ash weight, and Ca content of femur and 45Ca activity were decreased dose-dependently. In histological examination, changes of tibia and teeth were observed at the dose of 2mg P/kg and following dose increase these changes were more significantly. Ultraviolet line of Tc. appeared three in proximal metaphysis of tibia and dentin at the dose of 1mg P/kg and 2mg P/kg, but at the dose of 4mg P/kg and 8mg P/kg 2nd and 3rd line could not be detected. These result indicated that the effects of EDTPO on serum ingredients, long bone and teeth are initiated at the dose of 2mg P/kg, appeared at the dose of 4mg P/kg significantly, and at the dose of 8 mg P/kg severely. In addition, increase of the ratio of 45Ca specific activity between serum and bone observed at the dose of 4mg P/kg and 8mg P/kg suggested that these effects of EDTPO would be due to inhibitory action on mineralization.

Alkaline Phosphatase↗

[Distribution of N-isopropyl-p-[I-123]iodoamphetamine among the peripheral blood components: an in vitro study].

With the purpose to clarify dynamics of N-isopropyl-p-[I-123]iodoamphetamine (I-123 IMP) in the blood stream its binding to the peripheral blood components was determined by in vitro experiment. I-123 IMP was added to the peripheral venous blood obtained from healthy volunteers to be incubated for different length of time (0-30 min) at 37 degrees C. The blood was then separated into blood cells and plasma. From the latter platelet rich plasma were separated. Radioactivity in each blood component was counted in a well type scintillation counter respectively. To evaluate the affinity of I-123 IMP to red blood cell the component containing blood cells were washed repeatedly with salines. It was found that the fraction of radioactivity in the blood cell component was 68.0 +/- 6.3% (m +/- 1 S.D.), which was higher than that in the plasma (32.0% +/- 6.3%). The radioactivity in the platelet-rich plasma was only 1.7 +/- 1.1% of the total I-123 IMP activity. This percentage did not change by the incubation time. When Tc-99m DTPA was incubated with blood, radioactivity in the blood cell component was only 22.5%, which is further lowered by 32 +/- 2.1% after each washing to reach 6.8% after three times washing. In contrast the radioactivity of I-123 IMP in blood cell component remained as high as 31.1% after eight times washing. Almost constant fraction (8.20 +/- 0.57%) of radioactivity was freed into supernate by each washing. These findings suggest that a certain specific binding mechanism is involved in the binding of I-123 IMP to red blood cells.

Amphetamines↗

An improved procedure for quantitating mitochondrial DNA in cultured mammalian cells.

A new improved method that reproducibly measures small perturbations of mitochondrial DNA in populations of cells has been developed. It is based on first obtaining a cell count and then analyzing three aliquots of cells: one for total DNA per cell by fluorometry, one for total protein per cell and one for the amount of mitochondrial DNA per microgram of total cell DNA. To quantitate mitochondrial DNA, 0, 1, 2, and 3 nanograms of mouse mtDNA purified from a plasmid are added as internal standard DNA to four 1.0-microgram samples of purified total cell DNA containing an unknown amount of mitochondrial DNA (a sample set). Three sample sets are electrophoresed in an agarose gel devoid of ethidium bromide. Following Southern transfer to nitrocellulose and hybridization to purified 32P-labeled mouse mitochondrial DNA, an autoradiogram is prepared for use as a template to locate the mitochondrial DNA bands. These bands are cut out of the nitrocellulose filters, and their 32P-content is determined using a liquid scintillation counter. For each sample set, the counts per minute is plotted against the amount of mitochondrial DNA added. The plot is linear and the negative average of the values for the three intercepts on the x-axis yields the amount of nanograms of mitochondrial DNA per microgram of total cell DNA. The method is highly reproducible with a standard deviation of approximately 9 percent. The advantages of using this method over others that have been reported are discussed.

Animals↗

Adherence of Mycoplasma hyopneumoniae to cell monolayers.

This work was an attempt to develop an in vitro adherence model for Mycoplasma hyopneumoniae, using monolayers of human and porcine lung fibroblasts and porcine kidney cells. Mycoplasma hyopneumoniae grown in Friis mycoplasma broth was radiolabeled with 35[S]-methionine, washed, concentrated, and inoculated on the monolayers. After 15 minutes of centrifugation to facilitate adherence, monolayers were washed 3 times, dissolved with 0.1N NaOH, and suspended in scintillation liquid, and the radioactivity was determined in a liquid scintillation counter. Adherence, measured as a percentage of counts added, varied according to the mycoplasma strain and the cell line used. Comparison of strains J, 144L, and 232 of M hyopneumoniae revealed 7.5 +/- 5.9, 31.9 +/- 13, and 9.6 +/- 5% adherence to porcine kidney cells, respectively. Slightly different, but proportionally the same relationships were obtained with swine or human fibroblasts. Adherence was decreased slightly by repeated washings of the mycoplasma-treated cell monolayers; however, a plateau was reached, indicating irreversibility of the adherence process. Pretreatment of cell monolayers with nonlabeled organisms substantially blocked adherence by labeled organisms. Dilution of labeled organisms resulted in an increased proportion adhering. Therefore, it appears that the adherence was a receptor-dependent event. Treatment of the mycoplasmas with trypsin prior to the inoculation of monolayers resulted in a marked reduction in adherence. Treatment of the mycoplasmas with hyperimmune swine serum against M hyopneumoniae or normal swine serum resulted in 80 to 90% reduction of adherence; however, no inhibition occurred when mycoplasmas were treated with purified IgG from the hyperimmune serum.

Animals↗

The influence of methionine-5-enkephalin on calcium uptake by the bovine aortic media.

The influence of methionine-5-enkephalin (M-5-E), an endogenous opioid receptor agonist, on calcium uptake by bovine aortic media was investigated in vitro. 45Ca was used and radioactivity was counted in a beta scintillation counter. M-5-E increases Ca2+ uptake by the preparation. This action is inhibited by naloxone and that is proof that an opioid receptor is stimulated. A comparative study showed that phenylephrine, an alpha-adrenoceptor agonist, exhibits the same action as M-5-E, whereas morphine's action is negligible. Phenylephrine contracts the deendotheliazed ring of the bovine aorta, whereas M-5-E fails to do so. It is concluded that an opioid receptor was identified at the bovine aortic smooth muscle. This receptor is stimulated by M-5-E resulting in an increase of the extracellular Ca2+ entrance. Although no relationship was observed between the receptor and the contraction mechanism, a possible role of M-5-E in the maintenance of the vascular tone cannot be excluded.

Animals↗

[Effect of 2-naphthyl-3- (3,4-dimethoxy) phenyl-propenoic acid on the metabolism of arachidonic acid in rabbit platelets].

Washed rabbit platelets were incubated with [14C] arachidonic acid (AA sodium salt) after being exposed to 2-naphthyl-3-(3,4-dimethoxy) phenyl-propenoic acid (NM-PA), indomethacin (Ind) and imidazole (Imi) or control solvent. The metabolites of AA in rabbit platelets were separated with the system of chloroform: methanol: acetic acid: water (90:8:1:0.8) by thin layer chromatography and quantitated by liquid scintillation counter. The metabolism of AA was influenced by NMPA dose-dependently in the range of 0.05-0.5 mmol/L. The formation of thromboxane B2 (TXB2) and 12-hydroxy-5,8,10-heptadecatrienoic acid (HHT), the final metabolites in the pathway of cyclooxygenase-thromboxane synthetase, were decreased from 24 +/- 6.7 to 3.2 +/- 1.6% and 10.3 +/- 2.49 to 4.7 +/- 2.8%, respectively. Meanwhile, 12-hydroxy-5,8,10,14-eicosatetraenoic acid (12-HETE), the metabolite in the pathway of lipoxygenase, was increased from 11.9 +/- 1.7 to 34 +/- 5.6%. It is suggested that NMPA blocks the pathway of cyclooxygenase-thromboxane synthetase and then changes the direction of arachidonate metabolism in platelets, the activation of platelets is inhibited in this way.

Animals↗

Incorporation of C14-thymidine and C14-uridine nucleosides into nucleic acid molecules in the liver of mice exposed to trivalent arsenic in drinking water.

The effect of trivalent arsenic on the C14-thymidine and C14-uridine incorporation into the liver nucleic acid molecules was studied in groups of mice exposed to 3.3 mmol . l-1 sodium arsenite in drinking water for 2, 4, 8, 16 and 32 days. In each exposure group, 18 h before the animals were killed by decapitation, six to ten mice were injected with 0.1 ml of C14-thymidine or C14-uridine of an activity of 0.37 MBq. The radioactivity of liver DNA and liver RNA fractions isolated and separated by extraction from the mouse liver homogenate was measured on a Betazcint BZ 5,000 scintillation counter and expressed as cpm . mg-1 of isolated nucleic acids. Initially, the rates at which the labeled thymidine and uridine nucleosides incorporated into the nucleic acid molecules showed a marked drop below the control value, then started to rise again, continuing to gradually increase as the time of arsenic exposure was increasing; they attained the control value on day 8, and at the last interval of observation (i.e. at 32 days of exposure to arsenic) they reached, respectively, 143% and 127% of the control value. The initial drop in the rate of nucleoside incorporation is believed to reflect the general metabolic inhibition caused by arsenic; the subsequent enhancement of the process of incorporation is ascribed to the induction of enzymes that are involved in the development of a tolerance for arsenic.

Animals↗

Comparison of antigen uptake by peritoneal macrophages and veiled cells from the thoracic duct using isotope-, FITC-, or gold-labelled antigen.

Veiled cells (VC), isolated from the thoracic duct of irradiated lymphadenectomized mice (MLNX) or peritoneal macrophages (PM phi were incubated with isotope-labelled hen egg lysozyme (HEL), purified protein derivative (PPD) or keyhole limpet haemocyanin (KLH) in vitro. About 2-10 times less antigen was associated with VC than with PM phi when measured in a Philips well-type scintillation counter. Autoradiographs of these cells indicated that 3-10% veiled cells had silver grains associated with them in contrast to 20-95% of the PM phi, depending on the type of antigen studied. It was also shown, from the distribution curves for grains in individual cells, that VC contained smaller numbers of grains than PM phi. Transmission electron microscopy using KLH conjugated with colloidal gold and immunofluorescence microscopy using KLH-FITC confirmed the results obtained from autoradiographs. However, measurements of the uptake of KLH-FITC by individual VC and PM phi using flow cytometry indicated that antigen was associated with nearly all VC in vitro but in much smaller amounts than with PM phi. Both VC and PM phi were capable of presenting HEL to primed T lymphocytes in vitro. These results are discussed in relation to the function of VC as accessory cells compared with PM phi.

Animals↗

The influence of amikacin on digoxin uptake by certain rabbit tissues in vitro.

The influence of amikacin on digoxin uptake by various rabbit tissues was investigated in vitro. 125J-digoxin was used and radioactivity was counted in a gamma scintillation counter. Amikacin decreases digoxin uptake by the renal tissue and this action is probably due to a displacing effect. On the contrary, amikacin increases digoxin uptake by striated and cardiac muscle. It is suggested that the latter action is due to a vasodilating effect of the aminoglycoside antibiotic that favors the microcirculation of the above tissues.

Amikacin↗

[The pharmacokinetic study of 3H-alpha-2-butylhydroxybenzyl alcohol in rabbits].

A derivative of Gastrodigenin, alpha-2-butylhydroxybenzyl alcohol, has shown a high distribution in the mice brain and stronger pharmacologic effects than its parent compounds. With 3H-alpha-2-butylhydroxybenzyl alcohol as radioactive tracer, pharmacokinetic data were obtained after the intravenous administration of a single dose alpha-2-butylhydroxybenzyl alcohol to rabbits, according to a cross-over design. Then, serial plasma samples were taken from 1 min to 480 min and measured by liquid scintillation counter. The data were analyzed by IBMPC computer for the estimation of pharmacokinetic parameters and the judgement of compartment model with a program recently developed by ourselves. The results suggested that the pharmacokinetics of alpha-2-butylhydroxybenzyl alcohol accords with the two compartment open model based on either the comparison of the calculated theoretic value with measured concentration or F test for the model judgement, and that the process of distribution was quite rapid, and the elimination half-life (T1/2 beta) was 12 h, indicating a slow elimination process.

Animals↗

Evaluation of the opsonic capacity of core lipopolysaccharide antiserum of equine origin against smooth Escherichia coli 0111:B4, using macrophage chemiluminescence.

A study was performed to determine whether equine antiserum to core lipopolysaccharide (LPS) would enhance phagocytosis of smooth gram-negative (GN) organisms by equine macrophages. Five healthy adult horses (group A) were immunized with a bacterin prepared from the J-5 mutant of Escherichia coli 0111:B4 and Salmonella minnesota R595 to produce antibodies to core LPS. Five horses (group B) served as nonimmunized controls and were given physiologic saline solution instead of the rough mutant bacterin. Serum antibody titers to core LPS and to smooth E coli 0111:B4 were determined by indirect ELISA. Four serum pools were prepared: pool 1 = sera from horses in group B prior to immunization; pool 2 = sera from horses in group A prior to immunization (preimmune serum); pool 3 = sera from horses in group B, 7 days after the last saline injection; pool 4 = sera from horses in group A, 7 days after the last immunization (core LPS antiserum). The serum pools, either unheated or heated 30 minutes at 56 C, in 3 dilutions (1/50, 1/100, 1/500) were used to opsonize smooth E coli 0111:B4 in an assay of equine peritoneal macrophage chemiluminescence (CL). Peritoneal fluid was collected from clinically normal horses and the macrophages were purified by adherence to borosilicate glass scintillation vials. Each serum type and dilution was added to triplicate vials containing 10(7) colony-forming units of E coli 0111:B4. Luminol-dependent CL was measured with a liquid scintillation counter in the out-of-coincidence mode. Each serum dilution was tested in duplicate vials without bacteria to asses serum-induced nonspecific CL.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

[The evaluation of split renal function in the experimentally induced transient renal vein obstruction].

The split renal function was measured in order to evaluate the alteration of renal function secondary from transient renal vein obstruction. Three microcuries of I-131 Hippuran and 7 microCi of Tc-99m DTPA were injected into the femoral vein of adult male rats 30 min, 3 hr, 6 hr, 2 days and 1 week after the renal vein obstruction for 30 min. Blood samples were taken from the tail vein 5, 10, 15, 20 and 30 min after injection. Immediately after taking blood sample at 30 min, both kidneys and urinary bladder with ureters were taken. The radioactivities of Tc-99m and I-131 of these samples were measured respectively using a well type scintillation counter at the energy ranges of 140 KeV +/- 25% and 360 KeV +/- 25%. In the obstructed side, the blood clearances of I-131 Hippuran (ERPF) and Tc-99m DTPA (GFR) were decreased at 30 min and 6 hr after the release of left renal vein obstruction. Therefore the filtration fraction was not significantly changed. In conclusion, the single study using either Tc-99m DTPA or I-131 Hippuran may be enough to evaluate the alteration of split renal function in a model of renal vein obstruction, although the combined study using both agents is more preferable.

Animals↗

Palmitylation of the glycoprotein IIb-IIIa complex in human blood platelets.

The presence of covalently bound palmitic acid in fibrinogen receptors, glycoproteins (GP) IIb and IIIa, has been explored in human blood platelets. Membrane fractions were isolated from fresh blood platelets labeled with [9,10-3H]palmitic acid and then analyzed for radioactive proteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Protein bands were visualized by staining with Coomassie Brilliant Blue, excised, and counted in a liquid scintillation counter. The results indicate that membrane proteins with electrophoretic mobility corresponding to glycoproteins IIb and IIIa incorporate [9,10-3H]palmitic acid. The palmitylated glycoproteins IIb and IIIa were immunoprecipitated by specific anti-GP IIb and GP IIIa antisera. It is interesting to note that the palmitylation of these glycoproteins occurred rapidly in platelets activated with 0.5 unit of thrombin or 30 microM ADP. At the concentration used (100 micrograms/ml), cycloheximide did not inhibit incorporation of [3H]palmitate into the glycoproteins showing that this process is not dependent upon protein synthesis. The acyl moiety was resistant to denaturating detergents, delipidation with organic solvents, and hydrolyzable with hydroxylamine. In the case of membrane protein with the electrophoretic mobility of GP IIb, the radioactive label was significantly decreased after reduction with 2-mercaptoethanol. Final identification of GP IIIa as an acylated product in human platelets incubated with [9,10-3H]palmitic acid was provided by two-dimensional polyacrylamide gel electrophoresis. In contrast to GP IIb alpha, GP IIIa isolated by this method showed the presence of attached radioactive palmitic acid residues. Analysis by high performance liquid chromatography after methanolysis of the [3H]palmitate-labeled glycoproteins confirmed the fatty acid nature of the label. Palmitylation is a newly identified post-translational modification of the fibrinogen receptor which may play an important role in its interaction with the membrane and/or its biological function.

Adult↗

[Microbiologic in vitro comparison of plaque-inhibiting mouthwashes].

The colonization on rat molars of S. sobrinus OMZ 176, S. mutans OMZ 376, or a combination S. sanguis OMZ 9 and S. sobrinus OMZ 176 after a short exposition to various fluoride solutions and disinfectants was tested in vitro. The test solutions included Act, Candida, Veadent, sodium fluoride, amine fluoride, stannous fluoride, zinc fluoride hexetidine, stannous fluoride/amine fluoride solutions, chlorhexidine and water. The sterilized rat molars were immersed in the above test solutions for 60 seconds, then incubated with streptococci in broth for 8 hours, again dipped into the same test solutions for 60 seconds and reincubated for an additional 30 hours. The streptococcal suspension contained 14C labelled sucrose solutions. The deposits on the molars were dissolved in 6N potassium hydroxide during 16 hours. Finally, the beta rays emitted by the dissolved radiolabelled suspension were counted using a scintillation counter. The sodium fluoride containing solutions exerted no or a very limited effect on the bacterial deposits. In contrast to the other test solutions, sanguinaria extract (Veadent mouth rinse) mildly inhibited the S. sobrinus and S. mutans deposits, but plaque formation by the combination of streptococci was not hampered. Zinc fluoride/hexetidine, amine fluoride and stannous fluoride/amine fluoride solutions had a distinct and significant inhibitory effect on S. sobrinus and S. mutans deposits, but only a weak effect when mixed cultures were used for plaque formation. Chlorhexidine significantly inhibited the deposits of the three bacterial strains used in these experiments.

Animals↗

Direct determination of angiotensin-converting enzyme inhibitors in plasma by radioenzymatic assay.

We describe a simple, rapid, specific radioenzymatic assay for "CGS 16617," a new, potent inhibitor of angiotensin-converting enzyme (ACE; EC 3.4.15.1) in human plasma. This assay is based on the principle that the inhibitor (i.e., the drug) binds to the ACE in plasma and hence the amount of free ACE in plasma is inversely related to the amount of active inhibitor present. Free enzyme is reacted with a radiolabeled substrate, and the radioactive product is selectively extracted into the scintillation cocktail for quantification. Fivefold-diluted plasma samples are incubated with [3H]hippuryl-glycyl-glycine enzyme substrate at 37 degrees C for 30 min and the liberated [3H]hippuric acid is selectively extracted into scintillation cocktail. The radioactivity is counted in a liquid scintillation counter. Both within-run and between-run, the variability (CV) of the assay is less than 10%. As little as 200 ng of the drug per liter can be quantified in 50-microL plasma samples. The method can also be used to assay two other ACE inhibitors, pentopril and CGS 14831, demonstrating that the method can be readily adapted to any ACE inhibitor having a single active component in plasma. The ester prodrug pentopril can also be assayed after ester hydrolysis. This method is suitable for analysis of large numbers of samples in clinical laboratories for routine monitoring of the concentrations of active ACE inhibitors in blood.

Angiotensin-Converting Enzyme Inhibitors↗

Evaluation of the EDTA-washed diet for use in the experimental production of zinc deficiency in human subjects.

Ethylenediaminetetraacetate (EDTA) is known to bind zinc (Zn) and other metals. EDTA-washed soy protein-based diet has been extensively used as a dietary model for the production of Zn-deficiency in human subjects as well as in experimental animals. The present study was conducted to test this diet for possible contamination with EDTA (inhereted by washing procedure) and also to evaluate it for its acceptability and palatibility by human subjects. In the first experiment, EDTA-washed soy protein-based diets were prepared as previously described (repeated washing with EDTA-sodium salt followed by washing with deionized water and filteration), but 14C-EDTA was used. Scintillation counter analysis showed no traces of 14C in the prepared diet (100% of the 14C activity was recovered in the filtrate). In the second experiment, the diets were prepared using inactive EDTA, then fed to 10 volunteers for sensory evaluation. Results of this trial showed that the palatibility and acceptability of these diets are generally very poor. In conclusion, the use of EDTA-washing procedure did not result in any contamination of the soy assay protein with the EDTA. However, the present dietary model may not be used to induce zinc deficiency in human volunteers on a long term basis since the palatibility of the washed product is poor.

Diet↗

Comparison of indium 111 oxine-labeled platelet aggregation between sutured and argon laser-assisted vascular anastomoses.

The thrombogenicity of argon laser-assisted vascular anastomoses (LAVAs) was compared with that of sutured vascular anastomoses (SVAs) by measurement of platelet aggregation at the site of repair in a canine model. Sequential 1 cm longitudinal carotid and femoral arteriotomies (n = 80) or jugular and femoral phlebotomies (n = 80) were performed, with each vessel having two tandem, randomly positioned arteriotomies or phlebotomies separated by a 4 cm length of intact vessel. One incision was repaired by SVA with continuous 6-0 polypropylene sutures and the other by argon LAVA. For the laser fusions, argon laser energy was applied to the adventitial surface of the vessel with a 300 micron fiberoptic probe with 0.5 W power, 1100 joules per square centimeter energy fluence, and 150 second exposure per 1 cm length. The arterial and venous segments of SVAs and LAVAs and an equivalent length of normal vessel were harvested at 48 hours (n = 16, 16, 16), 2 weeks (n = 12, 12, 12), and 4 weeks (n = 12, 12, 12). Autologous indium 111 oxine-labeled platelets were injected intravenously 48 hours before removal of the vascular repairs and the radioactivity of the specimens was determined on removal with a NaI (T1) well-type scintillation counter. Anastomotic platelet adherence index (APAI) was calculated as the ratio of emissions of SVA or LAVA to normal reference vessel.(ABSTRACT TRUNCATED AT 250 WORDS)

Anastomosis, Surgical↗