Effect of harvesting and sorting on beta-1 integrin in canine microvascular cells.

BACKGROUND: The goal of seeding prosthetic conduits with endothelial cells (ECs) has focused attention on the role of EC adhesion molecules. Cell preparation techniques may affect adhesion molecule expression and graft seeding. METHODS: Using fluorescent antibody labeling and flow cytometric analysis, this study examined the effectsof monolayer detachment (scraping vs trypsinization), timing of immunolabeling (pre- vs postdetachment), gene transfection (transfected vs nontransfected), and cell selection (antibiotic vs fluorescence sorting) techniques on beta-1 integrin expression in canine microvascular EC (K9MVEC). RESULTS: Cell scraping resulted in a significantly higher beta-1 integrin mean fluorescence intensity than did cell trypsinization (P < 0.05). No difference was observed with immunolabeling prior to versus following monolayer harvesting. Gene transfection had no significant effect on beta-1 integrin expression. No advantage was observed between cell selection methods (P > 0.05). CONCLUSION: This study suggests that the monolayer harvesting technique employed has a significant impact on beta-1 integrin quantification by flow cytometric analysis. Furthermore, microvascular EC expression of beta-1 integrin was not adversely affected by gene transfection.

Influence of hormone replacement therapy on graft patency after femoropopliteal bypass grafting.

OBJECTIVE: Thromboembolic events are more frequent in women with established cardiovascular disease who are receiving hormone replacement therapy (HRT). The effect of HRT on the outcome of women undergoing infrainguinal bypass grafting is unknown. The purpose of this study was to estimate the influence of risk factors, in particular HRT, on the outcome of women undergoing femoropopliteal bypass grafting. METHODS: During a 5-year period (between 1993 and 1998), 131 femoropopliteal bypass graft procedures were performed in 106 women. The criteria prepared by the Ad Hoc Committee on Reporting Standards (Society for Vascular Surgery/North American Chapter of the International Society for Cardiovascular Surgery) were followed. Both univariate (Kaplan-Meier method) and multivariate analyses (Cox proportional hazards model) were used to determine the association among preoperative variables, graft patency, limb salvage, and survival. RESULTS: The average age of the patients was 66.4 years; 26% of the patients were receiving HRT. Indications for femoropopliteal bypass grafting were limb salvage (80%) and disabling claudication (20%). Autogenous vein was used in 48% of procedures, polytetrafluoroethylene (PTFE) in 49%, and PTFE-vein composite grafts in 3%. Distal popliteal anastomosis was above the knee in 52% and below the knee in 48%. Overall primary patency rate was 81% at 1 year, 65% at 3 years, and 56% at 5 years. Primary patency rates at 1, 3, and 5 years were 75%, 45%, and 23%, respectively, for HRT users and 84%, 72%, and 65%, respectively, for nonusers of HRT. Overall, cumulative 1- and 5-year limb salvage results were 96% and 92%, respectively, and long-term survival at 1, 3, and 5 years was 96%, 86% and 74%, respectively. With univariate and Cox regression analyses, HRT was identified as the only independent predictor of reduced primary graft patency (Kaplan-Meier method, log-rank test, P =.004; relative risk, 2.5; 95% CI, 1.3-4.8). Women receiving HRT who underwent a procedure with PTFE had the lowest primary graft patency rates (relative risk, 3.4; 95% CI, 1. 5-7.8; P =.006). CONCLUSIONS: Women undergoing femoropopliteal bypass graft procedures who are receiving HRT have significantly reduced primary graft patency rates. The risk of graft failure increases when prosthetic materials are used.

Odor of specific alcohol solutions selectively alters locomotor activity of alcohol-drinking P rats.

Evidence exists that the olfactory property of alcohol affects cerebral processes to influence the pattern of alcohol drinking in an experimental animal. In this study, the level of activity was quantified in the genetically selected alcohol-preferring P rats during exposure to the odor of 4 concentrations of alcohol. A Mini-Mitter transmitter was implanted intraperitoneally in each animal to record individual locomotor activity continuously, in terms of counts per unit time. In the first experiment, each rat was exposed between 1730 and 1930 h to the odor of 5%, 15%, 30%, or 60% alcohol placed in 2 petri dishes flanking both sides of each cage. The control condition was identical, except that the petri dishes were empty. In the second experiment, each rat was offered water and its individually preferred solution of alcohol in the drinking tubes during 1730 to 1930 h on alternate days of exposure to the odor of alcohol. During the odor condition alone, the 30% concentration of alcohol increased the activity counts significantly from the control level of 228.8 +/- 23.9 to 303.5 +/- 28.1 over the first 30 min of the 2-h test period. Plots of activity every 30 min for each alcohol solution, calculated as percent of control, also showed that the odor of 30% alcohol evoked significantly greater activity during the first and fourth intervals, in contrast to the exposure to 5%, 15%, and 60% alcohol, which failed to shift activity beyond the control level. When the P rats were exposed to 30% alcohol 24 h after 2 h of alcohol drinking, their profile of activity was concordant with that prior to alcohol drinking. These results demonstrate that the odor of a specific concentration of alcohol (i.e., 30%) selectively enhances the activity of the P rat. This could reflect an association of the sensory quality of alcohol with an anticipation of drinking. Finally, these experiments provide the first quantitative evidence that an olfactory cue associated with the unique odor of a specific solution of alcohol may determine the nature of the drinking response.

The mixed 5-HT 1A/2A receptor drug FG5938 suppresses alcohol drinking while enhancing feeding in P rats.

The neurotransmitter serotonin (5-HT) has long been implicated in the etiology of aberrant consumption of alcohol. Several compounds thought to possess a potential therapeutic value to counteract drinking have high affinities for 5-HT1A and 5-HT2A receptors in the brain. For example, amperozide and FG5865 significantly reduce the volitional intake of alcohol, without altering food intake, both in rats genetically predisposed or chemically induced to drink alcohol. The present study was undertaken in the alcohol-preferring (P) rat to determine whether an amperozide like drug. FG5938 (1-[4-(p-fluorophenyl)butyl]-4-(6-methyl-2-pyridinyl)-piperazine fumarate). exerts an action on the volitional drinking of alcohol as well as on the intakes of food and water. In 11 male P rats, the pattern of preference for different concentrations of alcohol was determined by an 11-day test for water vs. 3 to 30% alcohol solutions. After maximally preferred alcohol concentrations, i.e., 9 to 15% had stabilized for 4 days, saline or FG5938 was injected subcutaneously at 1600 and 2200 h in a dose of 2.5, 5.0, or 10.0 mg/kg over 4 consecutive days. Following treatment, preference testing for the same concentrations of alcohol was continued for 5 additional days. FG5938 caused a significant suppression in alcohol drinking in terms of both absolute g/kg and proportion to total fluid intake. During its administration, FG5938 also enhanced the ingestion of food and water of the P animals significantly, with the largest intake occurring on the initial day, while body weights increased. After FG5938 injections, food and water intakes returned to predrug levels. The saline control vehicle had no significant effect on the intakes of alcohol, food, or water of the P rats. Overall, these results show that FG5938 acts to attenuate alcohol preference while simultaneously increasing the ingestion of food paradoxically. To our knowledge, this is the first known drug to possess this unique property. Finally, these findings support the view that a compound having affinities to both 5-HT1A and 5-HT2A receptors may be useful as a therapeutic agent in the treatment of alcoholism.

Tomato juice, chocolate drink, and other fluids suppress volitional drinking of alcohol in the female Syrian golden hamster.

Although the hamster generally prefers alcohol at a level similar to that of the rat or mouse selectively bred to consume alcohol, the drinking hamster demonstrates neither physical dependence on alcohol nor elevated blood levels of alcohol, which are two typical criteria characterizing an animal model of alcoholism. The present investigation was designed to determine whether a third criterion of an animal model (i.e., consumption of high levels of alcohol in the presence of a palatable fluid, fulfilled by the P rat) would be met by the female Syrian golden hamster (Mesocricetus auratus). A standard 3-bottle preference test was undertaken in 6 female hamsters over an 11 day period, in which water was presented in one tube and, in a second tube, a v/v solution of alcohol which was increased in concentration from 3% to 50% on each day as follows: 3%, 5%, 7%, 9%, 12%, 15%, 20%, 25%, 30%, 40%, and 50%. Then each hamster was offered its individually determined, maximally preferred concentration of alcohol for 4-8 days, which was 20%, 25%, or 30% alcohol. The mean absolute intake of alcohol during this period was 17.9 +/- 1.1 g/kg per day, whereas the mean proportion of alcohol to total fluid was 0.68 +/- 0.05. Then over a 4-day interval, a solution of tomato juice, peach juice, mango juice, dextrose and a chocolate beverage (Ensure Plus), all made isocaloric to the alcohol solutions with dextrose, was placed in the third tube simultaneously with water and the individually preferred concentration of alcohol.(ABSTRACT TRUNCATED AT 250 WORDS)

Female Syrian golden hamster: drinking of high concentrations of ethanol aversive to other mammals.

The present experiments were designed to determine: 1) the pattern of preference for different concentrations of ethanol in the female Syrian golden hamster (Mesocricetus auratus), and 2) the influence of drinking ethanol on their intakes of food and total calories. A standard three-bottle preference test was undertaken in six female hamsters over an 11-day period in which water was offered together with ethanol, which was increased in concentration over 11 days from 3% to 50% as follows: 3%, 5%, 7%, 9%, 12%, 15%, 20%, 25%, 30%, 40%, 50%. Then, each hamster was offered its individually preferred concentration for a period of 8 days until the intake of ethanol had stabilized. During the preference testing for 3-25% solutions, the proportional intakes ranged between 0.6 and 0.8 whereas the mean absolute amount consumed per day increased from 2.3 to 16.1 g/kg at the 25% concentration. However, at the 50% concentration, ethanol drinking declined substantially to 8.7 g/kg per day. The overall mean percent concentration of ethanol preferred by the hamsters was 24.2 +/- 1.5%. During the following 8-day period when the maximally preferred concentration of ethanol of each hamster was offered with water, the mean intake of ethanol was 17.9 +/- 1.1 g/kg per day. Throughout the test sequence, the caloric intake of the animals was maintained in that calories obtained from food declined at the same rate as the calories obtained from ethanol in rising concentrations.(ABSTRACT TRUNCATED AT 250 WORDS)