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A new, automated assay for von Willebrand factor (vWF) activity has recently become commercially available (HemosIL vWF activity assay, Instrumentation Laboratories, Lexington, MA). We prospectively studied 61 specimens from 58 patients undergoing laboratory testing for suspicion of von Willebrand disease with this new method, in comparison with the established ristocetin cofactor method. Assays for factor VIII and vWF antigen were also performed using an established method on an MDA-180 coagulation analyzer (bioMerieux, Durham, NC) and a new method on an ACL TOP coagulation analyzer (Instrumentation Laboratories). Blood types were determined. The results showed no significant difference between the assays for factor VIII (mean, 97% for MDA-180 and ACL TOP; P = .494) or vWF antigen (mean, MDA-180, 109%; ACL TOP, 111%; P = .766). The mean result for the ristocetin cofactor assay was 106% vs 93% with the automated vWF activity ( P = .007). The automated activity assay was 100% sensitive and 86% specific for detecting vWF abnormalities and seems to be a suitable screening test. Abnormal results should be followed up with a ristocetin cofactor activity assay for confirmation. Further study is recommended to confirm these conclusions.

Background: The goal of this study was to investigate the effect of fatty acid ethyl esters (FAEE), nonoxidative metabolites of ethanol, on platelet function. We hypothesized that FAEE increase the risk of bleeding by producing an alteration in platelet membrane structure or function. Methods: Isolated human platelets incubated with FAEE were prepared and multiple assays for platelet activation were performed; β-thromboglobulin release from platelet granules, platelet aggregation, arachidonate release from phospholipids, and intracellular cyclic AMP (cAMP) levels. We examined also the combined effect of epinephrine and FAEE on platelet aggregation. Results: FAEE induced platelet shape change, release of α granules and release of arachidonate from phospholipids without an increase in eicosanoid production and decreased cAMP levels. The platelets did not aggregate in response to FAEE alone, but did shorten the time to maximum aggregation with epinephrine. Conclusion: These studies show that FAEE potentiate platelet activation but do not induce aggregation, presumably because they do not stimulate thromboxane A2 production.

Background: Fatty acid ethyl esters (FAEE) are nonoxidative ethanol metabolites shown to produce toxic effects in the liver and pancreas in vivo and in vitro. Because alcohol-induced chronic pancreatitis is associated with mutations in the gene responsible for cystic fibrosis (CFTR), we hypothesized that CFTR dysfunction leads to increased levels of these toxic nonoxidative ethanol metabolites following alcohol administration. Methods: Cystic fibrosis (CF) and wild-type (WT) mice were injected intraperitoneally with 1, 2, or 3 g/kg of 50% ethanol. Mice were sacrificed and the liver and pancreas removed for FAEE analysis. Results: The mean FAEE concentration (pmol/g) detected in the liver of cftr−/− mice following injection with 2 g/kg of ethanol was significantly greater than the amount detected in WT (p < 0.005). A similar trend in FAEE concentration was seen in the pancreas, but the difference was not statistically different. In both the liver and pancreas, analysis of individual FAEE species demonstrated a selective increase in ethyl oleate. Conclusion: These data show an association between CFTR dysfunction and qualitative and quantitative changes in FAEE in liver and pancreas upon ethanol exposure.

Fatty acid ethyl esters (FAEE), esterification products of fatty acids and ethanol, have been shown to be mediators of ethanol-induced cell injury and their presence in the blood and tissues is a marker of ethanol intake. Recently, it has been shown that FAEE are produced within seconds of infusion of ethanol into the heart, when using a protocol similar to that used for myocardial ablation. This raises the possibility that the mechanism for the death of myocytes in cardiac ablation involves the generation of toxic FAEE. It has also been recently demonstrated that chronic alcoholics have a high concentration of a specific FAEE species—ethyl oleate. The use of the serum ethyl oleate concentration may be helpful in differentiating binge drinkers from chronic alcoholics.

Background: Fatty acid ethyl esters (FAEEs) are nonoxidative metabolites of ethanol. FAEEs are found in liver, pancreas, and adipose tissues up to 24 h after consumption of ethanol, and on that basis, they are potentially useful markers for ethanol intake. In this study with rats, we investigated the efficacy of using FAEEs in liver and in adipose tissue as postmortem markers for premortem ethanol ingestion.Methods: An animal study was conducted in which test rats received injections of ethanol and control rats received injections of normal saline. The rats were killed 2 h after the injections. The bodies of the animals were stored at 4 °C up to 12 h, and samples of liver and adipose tissues were collected at different time intervals and processed for FAEE quantification. In another set of experiments, the rats received injections and were killed as described above, but bodies of animals from both groups were stored at 4, 25, or 37 °C for up to 72 h, and liver samples were collected and processed for FAEE quantification.Results: FAEEs were detected up to 12 h after death in liver and adipose tissue samples from the bodies of ethanol-treated animals stored at 4 °C; negligible amounts were detected in the bodies of animals that received normal saline. Adipose tissues contained higher amounts of FAEEs than liver, as well as more species: eight FAEE species in adipose tissue and five in liver tissue. Higher concentrations of FAEEs were detected in livers of treated animals stored at 25 °C for up to 48 h than in livers of controls stored under the same conditions.Conclusions: For at least 12 h after death, FAEEs in liver and adipose tissues are useful postmortem markers of premortem ethanol ingestion.

Objective: There are various demographic and educational factors that affect the academic performance of undergraduate medical students. The objective of this study is to identify these factors and correlate them to the GPA of the students. Methods: A cross-sectional study design utilizing grade point averages (GPAs) of two cohorts of students in both levels of the pre-clinical phase. In addition, self-administered questionnaire was used to evaluate the effect of these factors on students with poor and good cumulative GPA. Results: Among the various factors studied, gender, marital status, and the transportation used to reach the faculty significantly affected academic performance of students. Students with a cumulative GPA of 3.0 or greater significantly differed than those with a GPA of less than 3.0 being higher in female students, in married students, and type of transportation used to reach the college. Factors including age, educational factors, and type of transportation used have shown to create a significant difference in GPA between male and females. Conclusion: Factors such as age, gender, marital status, learning resources, study time, and the transportation used have been shown to significantly affect medical student GPA as a whole batch as well as when they are tested for gender.

Aims: Skeletal muscle appears to be susceptible to chronic and acute excess alcohol intake, giving rise to alcoholic myopathy, a common disease among alcoholics. Fatty acid ethyl esters (FAEE), non-oxidative metabolites of ethanol, have been shown to be toxic to cells in vitro and in vivo. We hypothesized that accumulation of FAEE in skeletal muscle could contribute to the devel- opment of alcoholic myopathy. Methods: Male wistar rats were treated either with 75 mmol ethanol/kg body weight or saline, in the fed state or starved for 1 or 2 days before administration. Rats were thus divided into the following groups: fed-saline (n = 8); fed-ethanol (n = 8); starved 1 day, saline (n = 8); starved 1 day, ethanol (n = 9); starved 2 days, saline (n = 7); and starved 2 days, ethanol ( n= 8). At the end of the incubation, skeletal muscles (abdominal and gastrocnemius), liver, and heart were isolated and processed for FAEE isolation and analysis by gas chromatography-mass spectrometry (GC-MS). Results: Total mass of FAEE in the muscles was much greater than that found in the liver and the heart. In general, the animals that were fasted for 1 day and received ethanol had the highest FAEE levels among the three groups of animals. The major ethyl ester species in all cases were ethyl 16:0, ethyl 18:0, ethyl 18:1 n-9, and ethyl 18:2 n-6. Ethyl 20:4 n-6 and ethyl 22:6 n-3 were also present, except in the fasted 1-day group, where ethyl 22:6 disappeared, though it reappeared in the fasted 2-day group. Conclusion: These findings demonstrate that skeletal muscles contain high levels of FAEE that are synthesized in the body after ethanol exposure. The concentration of FAEE in skeletal muscle in this study was very sim- ilar to FAEE concentration in the liver. This differs from previous studies suggesting a low concentration of skeletal muscle FAEE with ethanol exposure.

Context.—Technologic advances affecting analyzers used in clinical laboratories have changed the methods used to obtain many laboratory measurements, and many novel parameters are now available. The effects of specimen transport through a pneumatic tube system on laboratory results obtained with such modern instruments are unclear. Objective.—To determine the effects of sample transport through a pneumatic tube system on routine and novel hematology and coagulation parameters obtained on state-of-the-art analyzers. Design.—Paired blood samples from 33 healthy volunteers were either hand delivered to the clinical laboratory or transported through a pneumatic tube system. Results.—No statistically significant differences were observed for routine complete blood cell count and white cell differential parameters or markers of platelet activation, such as the mean platelet component, or of red cell fragmentation. When 2 donors who reported aspirin intake were excluded from the analysis, there was a statistically, but not clinically, significant impact of transport through the pneumatic tube system on the mean platelet component. There were no statistically significant differences for prothrombin time, activated partial thromboplastin time, waveform slopes for prothrombin time or activated partial thromboplastin time, fibrinogen, or fibrin monomers. Conclusions.—Although further study regarding the mean platelet component may be required, transport through a pneumatic tube system has no clinically significant effect on hematology and coagulation results obtained with certain modern instruments in blood samples from healthy volunteers.

The consumption of alcohol is known to have both positive and negative effects on the functioning of the cardiovascular system in general, and on platelet function in particular. Fatty acid ethyl esters (FAEEs) are non-oxidative metabolite of ethanol that may mediate the ethanol effect on platelet function leading to either bleeding or clotting. The aim of the current study was to investigate the synthesis, uptake, and hydrolysis of FAEEs by human platelets. Isolated platelets were incubated with ethanol for various times, and FAEE synthesis were measured by gas chromatography mass-spectrometry (GC-MS). In addition, platelets were incubated with 14C-ethyl oleate, and FAEE uptake and hydrolysis were measured. There was significant synthesis of FAEEs by human platelets within 30 min of exposure to ethanol. The major FAEE species formed by human platelets exposed to ethanol were ethyl palmitate and ethyl stearate. FAEE uptake by human platelets showed maximum uptake by 60 s. The majority of FAEEs (50–80%) incorporated into platelets remained intact for up to 10 min. FAEE hydrolysis led to an increase in free fatty acids, with minimal subsequent esterification of the free fatty acids into phospholipids, triglycerides, and cholesterol esters. These studies show that FAEEs, non-oxidative metabolite of ethanol, can be incorporated into, synthesized, and hydrolyzed by human platelets.

Several epidemiologic studies have shown that moderate intake of alcohol is associated with a lower risk of cardiovascular disease (CVD), but the mechanism is not fully elucidated. One of the proposed mechanisms of the protective effect of moderate alcohol intake is its beneficial effect on hemostasis. The aim of this review is to summarize the effect of ethanol intake on platelet aggregation and activation, coagulation factors including von Willebrand factor (vWF), and the fibrinolytic system. With regard to the effect of alcohol on platelet function, evidence in the literature suggests both platelet activation and platelet inhibition by ethanol. A unifying hypothesis is that platelets are partially activated by ethanol, with partial degranulation allowing for continued circulation of platelets with impaired function. Evidence also exists showing that ethanol intake decreases fibrinogen, factor VII, and vWF levels. In addition, alcohol intake has been found to increase fibrinolysis by increasing tissue plasminogen activator activity. The effect of ethanol on platelets, coagulation factors, and the fibrinolytic system is likely to contribute to protection against CVD.

Leptin is a nutritionally regulated adipocyte-derived cytokine. Previous studies in obese patients have demonstrated increased inflammatory markers and increased platelet aggregation in association with leptin. However, the effects of leptin administration on markers of inflammation and platelet aggregation in a human model of undernutrition have not previously been studied.The objective of the study was to investigate markers of inflammation, platelet activation, and platelet aggregation in a model of caloric deprivation and increased leptin sensitivity.This study was a randomized, placebo-controlled study conducted between November 2002 and November 2003.The study was conducted at an inpatient care setting at the General Clinical Research Center.Twenty healthy, young (18-35 yr old), normal-weight (body mass index, 20-26 kg/m2) women were recruited from local advertisements. No subjects withdrew due to adverse effects.The effects of physiological recombinant methionyl human leptin or identical placebo administration were investigated over a 4-d fast.The primary outcome measures for this study were C-reactive protein (CRP) and indices of platelet activity.Leptin administration prevented the fasting-induced decline in leptin (P0.05 vs. placebo at each time point). Leptin administration increased CRP (6.3 +/- 2.4 vs. 0.7 +/- 0.3 mg/liter; P = 0.04), circulating P-selectin (11.6 +/- 10.2 vs. -28.9 +/- 15.6 ng/ml; P = 0.04), and induction of platelet aggregation (5.8 +/- 2.6 vs. -2.7 +/- 2.9%, P = 0.04, percent maximum platelet aggregation) relative to placebo administration (change in leptin vs. change in placebo, respectively, for each variable). Leptin tended to increase serum amyloid A [0.1 +/- 0.2 vs. -0.3 +/- 0.1 log10 (ng/ml); P = 0.07], and the changes in serum amyloid A and CRP were highly correlated (r = 0.83; P0.0001). No changes in TNFalpha, IL-6, IL-10, plasminogen activator inhibitor-1, haptoglobin, intercellular adhesion molecule, or vascular cell adhesion molecule were seen between the groups.Our data provide evidence that physiological leptin administration stimulates inflammatory and platelet responses in humans during caloric deprivation.

This chapter discusses the relationship of alcohol and fatty acid ethyl esters. Fatty Acid Ethyl Esters (FAEE) are nonoxidative ethanol metabolites that are esterification products of fatty acids and ethanol. FAEE directly contribute to cell injury induced by ethanol abuse, but the exact mechanism by which FAEE mediate this toxic effect remains to be discovered. Most of the enzyme activities found to be catalytic for FAEE synthesis are esterification enzymes that link fatty acid or fatty acyl-CoA to ethanol. However, there are a number of reports that suggest that the hydrolysis of fatty acids from a more complex lipid molecule (such as phospholipid and triglyceride) is the rate limiting step in the formation of FAEE, and that the esterification step is presumably either nonrate-limiting or nonenzymatic. FAEE are of growing importance in the toxicity induced by excess ethanol intake. There is an increasing evidence that FAEE mediated cytotoxicity and FAEE are proving to be a robust marker of ethanol intake in a variety of cells and tissues, and in the blood. The use of FAEE to monitor ethanol intake or to identify individuals as chronic alcoholics should continue to strengthen as more clinical studies are completed.

Fatty acid ethyl esters (FAEEs) are nonoxidative ethanol metabolites that have been implicated as mediators of alcohol-induced organ damage. FAEEs are detectable in the blood after ethanol ingestion, and on that basis represent markers of ethanol intake. FAEEs have also been quantitated in human liver and adipose tissue and have been shown to be postmortem markers of premortem ethanol intake. A substantial difference in FAEE concentration was found in liver and adipose tissue of patients with detectable blood ethanol at the time of autopsy vs those with no detectable blood ethanol, who were either chronic alcoholics or social drinkers. Most currently available diagnostic markers for chronic alcoholism have limited clinical utility. Data in this report demonstrate that the amount or type of FAEEs can be used to differentiate a chronic alcoholic from an episodic heavy drinker (binage drinker) at or near peak blood ethanol concentrations and approximately 24 hours after discontinuation of ethanol. Thus, FAEEs are markers of ethanol intake in blood and tissues and can be useful in distinguishing chronic alcoholics from binge drinkers.

Fatty acid ethyl esters (FAEEs) are cytotoxic nonoxidative metabolites of ethanol that are produced by esterification of alcohol and fatty acids (1)(2)(3). After ethanol intake, FAEEs are found mainly in the liver, pancreas, heart, and brain, which are the major organs adversely affected by ethanol intake (4)(5). FAEEs are detectable in the blood for up to 24 h after consumption of ethanol to at least 600 mg/L (6)(7)(8), making the presence of FAEEs in blood a useful marker for ethanol intake. This investigation presents the results of studies on the effects of collection tube, storage time, and storage temperature on FAEE concentrations in blood, with the goal of creating a reliable clinical assay for serum and plasma FAEE quantification. Four volunteers participated in this study. Each subject was given a weight-adjusted amount of 100-proof vodka mixed with fruit juice in a 1:3 ratio (8). The vodka-juice beverage was divided into nine equal aliquots, which were administered every 10 min over a 90-min time period. After the last aliquot was drunk, blood samples were collected in four different 5-mL Vacutainer Tubes from each subject to assess the influence of the collection tube …