The prevalence of hemoglobin Tacoma in Finland detected by HbA1c capillary electrophoresis.

Previous studies have identified occasional cases of heterozygous Hb Tacoma in areas that have attracted Finnish immigrants, especially in Sweden and North America, but large studies of this slightly unstable beta variant in vitro have not been carried out. Here we determined the prevalence of hemoglobin variants across Finland. A total of 5059 samples from 11 different hospital districts were analyzed using HbA1c capillary electrophoresis and reviewed for atypical profiles (HbA1c, Capillarys 3 Tera, Sebia). 38 heterozygous Hb Tacoma cases were found (0.75%). The prevalence was highest in South Ostrobothnia (2.0%), located in western Finland, and second highest in the neighboring provinces (1.0-1.4%), but only two districts were devoid of variants. Heterozygous Hb Tacoma was confirmed by genetic testing (NM_000518.5(HBB):c.93G > T (p.Arg31Ser)). In addition, five other variants were found, suggestive of HbE, Hb Helsinki (two cases) and an alpha variant, as interpreted from the electropherograms. The fifth variant, belonging to the South Ostrobothnian cohort, remained unknown at the time of the initial analyses, but was later interpreted as homozygous Hb Tacoma and confirmed by hemoglobin fraction analysis (Hemoglobin(E), Capillarys 3 Tera). In a subsequent retrospective study of the electropherograms of routine HbA1c diagnostics, altogether nine homozygous Hb Tacoma cases were identified in South Ostrobothnia. While heterozygous Hb Tacoma is usually an incidental finding, it interferes with several HbA1c assays. The present study is the first demonstration of homozygous Hb Tacoma. The clinical presentations of homozygous Hb Tacoma are not known and need to be addressed in future studies.

Serum Calprotectin, a Marker of Neutrophil Activation, and Other Mediators of Inflammation in Response to Various Types of Extreme Physical Exertion in Healthy Volunteers.

PURPOSE: While extreme physical exertion is known to induce changes in the status of inflammation comparisons of the responses for various mediators of inflammation after acute bouts of high-intensity exercise have been limited. SUBJECTS AND METHODS: We examined the responses in serum levels of novel inflammatory proteins, calprotectin, suPAR, CD163, and pro- and anti-inflammatory cytokines in 12 physically active volunteers (10 men, 2 women, mean age 37±14 years) before and after completing various types of extreme physical exertion (marathon run, half-marathon run or 24-h cross-country skiing). For comparisons, the levels of the biomarkers were also measured at rest in 30 healthy controls (25 men, 5 women, mean age 42 ± 12 years) with low or sedentary activity. RESULTS: Extreme physical exertion induced significant increases in serum calprotectin (p < 0.0005), suPAR (p < 0.01), CD163 (p < 0.05), IL-6 (p < 0.0005), IL-8 (p < 0.01) and IL-10 (p < 0.0005) (pre- vs 3h-post-exercise). These responses were found to normalize within 48 hours. While the increases in blood leukocytes were of similar magnitude following the different types of exercise, markedly more pronounced responses occurred in serum TNF-α (p < 0.01), IL-8 (p < 0.01) and CD163 (p < 0.05) in those with more intense activity. In 3-h post-exercise samples significant correlations were observed between serum calprotectin and IL-6 (rs = 0.720, p < 0.01), IL-10 (rs = 0.615, p < 0.05), TNF-α (rs = 0.594, p < 0.05), suPAR (rs = 0.587, p < 0.05) and blood leukocytes (rs = 0.762, p < 0.01). CONCLUSION: The present results suggest distinct exercise-intensity dependent changes in mediators of inflammation (including calprotectin, suPAR and CD163) following extreme physical exertion. Our findings indicate that there is a major reversible impact of high-intensity physical exertion on the status of inflammation.

Acute Changes in Inflammatory Biomarker Levels in Recreational Runners Participating in a Marathon or Half-Marathon.

BACKGROUND: Strenuous physical activity activates the participant's immune responses; however, few studies exist, observing exercise-induced simultaneous changes in mediators of inflammation. METHODS: We examined individual responses in soluble urokinase-type plasminogen activator receptor (suPAR), a marker of immune activation, soluble endocytic receptor for haptoglobin-hemoglobin complexes (CD163), a marker of monocyte-macrophage activation, C-reactive protein (CRP), and pro- and anti-inflammatory cytokines from blood samples drawn at baseline, at 3- and 48-h post-races from recreational runners who successfully completed the marathon (199 ± 8 min, n = 4) or half-marathon (132 ± 4 min, n = 4) run. For comparisons, biomarkers reflecting muscle, heart, kidney, and liver functions were measured. RESULTS: Significant 3-h post-race increases occurred in levels of suPAR (p < 0.01), CD163 (p < 0.05), white blood cells (p < 0.001), pro-inflammatory cytokines, interleukin-6 (IL-6) (p < 0.001), IL-8 (p < 0.05), and anti-inflammatory cytokine IL-10 (p < 0.05), whereas tumor necrosis factor-α (TNF-α) and transforming growth factor-ß (TGF-ß) remained relatively stable. Full-marathon running lead to more pronounced increases in suPAR, CD163, IL-8, and IL-10 than half-marathon running. In addition, 3-h post-race increases of all these parameters correlated significantly with changes in serum TNF-α and cortisol. The 48-h levels of serum suPAR and both pro- and anti-inflammatory cytokines had decreased to baseline levels, whereas CRP, a marker of acute phase response, increased in those with the most prominent IL-6 and IL-10 elevations in their preceding samples. The highest suPAR, CRP, IL-6, TNF-α, IL-10, and cortisol levels were noted in the individual with the most severe post-race fatigue. CONCLUSIONS: Prolonged running increases mediators of inflammation in an exercise-dose-dependent manner which should be considered in the assessment of health status of physically active individuals after recent acute bouts of strenuous exercise.

Individual responses in biomarkers of health after marathon and half-marathon running: is age a factor in troponin changes?

Although strenuous physical activity is known to cause notable perturbations in blood chemistries, only few studies exist observing exercise-induced simultaneous changes in biomarkers of health status. We compared markers of muscle, cardiovascular, renal, hepatic and inflammatory status at baseline and at 3-h and at 48-h postrace in recreational runners who successfully completed either a marathon (mean age 27 ± 13 years, finishing time 199 ± 8 min, n = 4) or half-marathon (mean age 38 ± 13 years, finishing time 131 ± 6 min, n = 6) race. Significant postrace changes occurred in myoglobin (p < .001), creatinine kinase (p < .01), CK-MB-mass (p < .01), high sensitivity troponin I (p < .05), high sensitivity troponin T (p < .05), brain natriuretic peptide (p < .001), creatinine (p < .01), aminotransferase enzymes (p < .001 for AST and p < .01 for ALT), uric acid (p < .001) cortisol (p < .01), C-reactive protein (p < .05), leukocytes (p < .001), haematocrit (p < .05) and mean corpuscular volume (p < .01). In comparison between the two types of exercise, marathon running lead to more pronounced responses in myoglobin, CK-MB-mass, ALT, AST, lactate and phosphate. Notable elevations in troponin levels were observed only in young participants (<30 years), most strikingly in those ≤20 years of age. The data indicates that prolonged running leads to distinct biomarker alterations, which should be considered in the assessment of health status after recent acute bouts of strenuous exercise. The observations suggesting more pronounced cardiac troponin responses in young individuals warrant further studies in larger populations.

Clinical and Laboratory Responses of Cross-Country Skiing for a 24-H World Record: Case Report.

The physiological consequences of ultra-endurance cross-country skiing in cold conditions are poorly known. We report here clinical, echocardiographic and laboratory findings from a 41-y old male elite skier in a world record trial for 24-h skiing. The athlete completed a total of 406.8 km outdoors with the temperature ranging between -24°C and -5°C during the 24-h period. Post exercise, notable increases from baseline values were observed in myoglobin (50-fold), creatinine kinase (30-fold) and proBNP (6-fold), whereas troponin T or troponin I levels remained unchanged. At baseline, echocardiographic findings showed cardiac hypertrophy and after skiing, a 5% reduction of left-ventricular end-diastolic dimension. Increases in markers of kidney (creatinine) and liver function (alanine aminotransferase), serum uric acid, C-reactive protein and white blood cell counts were also noted. In addition, electrolyte disturbances including hyponatremia, hypophosphatemia and hypocalcaemia were noted during the follow-up. The data indicates that a prolonged period of high-intensity skiing leads to muscle, heart and kidney affection and activation of inflammation even in an experienced elite skier. The observed health effects underscore the need for strict medical surveillance of participants in extreme sports with long duration. Key pointsAn elite athlete was able to ski over 400 km during 24 hours with an outdoor temperature ranging between -5 °C and -24 °C.Several postrace abnormalities occurred in biomarkers of muscle, heart, kidney, liver and inflammation status.Serum troponins, specific markers of myocardial cell damage, remained stable.The report supports careful medical surveillance of participants in extreme sports with long duration.

Impacts of common factors of life style on serum liver enzymes.

AIM: To investigate the impacts of gender, age and factors of life style (alcohol, overweight, coffee and smoking) on serum liver enzymes. METHODS: Serum alanine aminotransferase (ALT) and gamma-glutamyltransferase (GGT) were measured from 6269 apparently healthy individuals (2851 men, 3418 women, mean age 45 ± 12 years, range 25-74 years) in a national cross-sectional health survey. All subjects underwent detailed clinical examinations and interviews including the amount and pattern of alcohol use, coffee consumption and smoking habits. RESULTS: In this population with a mean ± SD alcohol consumption of 65 ± 105 g/wk and body mass index (BMI) of 26.1 ± 4.3 kg/m(2), both ALT and GGT were significantly influenced by alcohol use (P < 0.001) and BMI (P < 0.001), whereas smoking increased only GGT (P < 0.001). A significant effect of age on ALT was seen in men (P < 0.001) whereas not in women. Significant two-factor interactions of alcohol use in men were observed with age (ALT: P < 0.01; GGT: P < 0.001) and BMI (GGT: P < 0.05). For ALT, a significant interaction also occurred between BMI and age (P < 0.005). In contrast, women showed significant interactions of alcohol use with BMI (GGT: P < 0.05), smoking (GGT: P < 0.001), and coffee consumption (GGT: P < 0.001). CONCLUSION: Life-style associated changes in liver enzymes may reflect health risks, which should be considered in the definition of normal limits for liver enzymes.

Serum soluble urokinase plasminogen activator receptor in alcoholics: relation to liver disease severity, fibrogenesis, and alcohol use.

BACKGROUND AND AIM: Heavy alcohol consumption may lead to development of liver disease and the need for non-invasive parameters for detecting those at risk is widely acknowledged. METHODS: We measured serum soluble urokinase-type plasminogen activator receptor (suPAR) levels from 63 patients with alcoholic liver disease (ALD), 57 heavy drinkers without apparent liver disease, and 39 controls who were either moderate drinkers or abstainers. RESULTS: The highest serum suPAR concentrations were detected in patients with ALD (P < 0.001) showing high diagnostic accuracy in differentiating ALD patients from heavy drinkers without liver disease (area under curve 0.921, P < 0.001). Levels of suPAR correlated positively with serum markers of fibrogenesis (aminoterminal propeptide of type III procollagen and hyaluronic acid) (P < 0.001), with clinical (combined clinical and laboratory index P < 0.01) and morphological (combined morphological index P < 0.05) indices of liver disease severity and with the stage of fibrosis (P < 0.01). The suPAR concentrations were also elevated in heavy drinkers when compared with healthy controls (P < 0.001). CONCLUSION: The data indicate that serum suPAR concentrations are increased as a result of heavy alcohol consumption and further with development of ALD, showing a good diagnostic performance in detecting those with liver disease. The association with the histological severity of ALD and correlation with fibrosis indicates potential of serum suPAR also as a prognostic marker in ALD.

Comparison of ethyl glucuronide and carbohydrate-deficient transferrin in different body fluids for post-mortem identification of alcohol use.

AIMS: Alcohol abuse is a major risk factor for premature death. Confirming the role of alcohol consumption in cause-of-death investigations has, however, remained difficult, due to lack of reliable biomarkers. METHODS: We compared ethyl glucuronide (EtG) and carbohydrate-deficient transferrin (CDT) assays from serum, urine, cerebrospinal fluid and vitreous humor in a forensic autopsy population with either a positive (n = 38) or negative (n = 22) history of alcohol abuse based on detailed medical and police records and forensic toxicological investigations. RESULTS: A positive blood alcohol concentration (median 1.15‰, range 0-3.3‰) was found in 26/38 (68%) of the cases with a documented history of alcohol abuse. EtG concentrations (mean ± SD) in urine (339 ± 389 mg/l, P < 0.001), vitreous humor (4.2 ± 4.8 mg/l, P < 0.001), serum (6.9 ± 8.9 mg/l, P < 0.01) and cerebrospinal fluid (1.7 ± 2.7 mg/l, P < 0.01) were significantly higher among the cases with a positive history of alcohol use than those in the alcohol-history negative group, whereas in corresponding comparisons CDT was significantly different only in cerebrospinal fluid (4.3 ± 2.1 vs. 2.3 ± 0.6%, P < 0.05). The highest sensitivities (92%) in detecting ante-mortem alcohol use were obtained for urine and vitreous humor EtG assays. CONCLUSION: Our data indicate that measurements of EtG in urine or vitreous humor show the highest diagnostic accuracies in post-mortem investigations of excessive alcohol consumption and can be recommended for routine applications.

Individual and joint impacts of ethanol use, BMI, age and gender on serum gamma-glutamyltransferase levels in healthy volunteers.

Excessive ethanol consumption, obesity and increasing age may all lead to increased serum levels of gamma-glutamyltransferase (GGT) enzyme, which plays a key role in the metabolism of extracellular reduced glutathione. However, as yet, the interactions between the various modulators of GGT activities have remained poorly defined. We analyzed data from 15,617 apparently healthy individuals (7254 men and 8363 women, mean age 46 ± 13 years, range 25-74 years) who participated in a national cross-sectional health survey in Finland between 1997 and 2007. All subjects underwent detailed clinical examinations and interviews, including the amount of ethanol use and smoking habits. GGT levels were measured from all participants, and the individual and joint impacts of the different study variables on GGT levels were assessed. Significant individual effects were noted for ethanol use (p < 0.001), body mass index (BMI) (p < 0.001), age (p < 0.001) and smoking (p < 0.001). In men, significant two-factor interactions occurred between ethanol use and age (p < 0.020). Among those over 40 years of age, ethanol consumption was found to be a stronger determinant of increased GGT levels than in men below 40 years, whereas in the latter age group, BMI was found to predominate. In women, a significant two-factor interaction occurred between ethanol and BMI (p = 0.010), whereas it did not with ethanol use and age. The data underscores the role of ethanol consumption and age as major determinants of increased GGT levels in men, whereas in women, a relatively stronger impact was noted for ethanol intake and BMI. In light of the ability of GGT enzyme to modulate crucial redox-sensitive functions, the present findings also support the use of GGT as a biomarker of oxidative stress.

Dose- and gender-dependent interactions between coffee consumption and serum GGT activity in alcohol consumers.

AIMS: Coffee consumption has been recently linked with decreased blood gamma-glutamyltransferase (GGT) activities and protection from alcoholic liver disease. To explore the relationship and dose response, we assessed the impacts of coffee and alcohol intake on serum GGT activity in apparently healthy men and women with varying levels of coffee and alcohol consumption. METHODS: Data on coffee, alcohol consumption and serum GGT activities were collected from 18,899 individuals (8807 men and 10,092 women), mean age 48 years, range 25-74 years, who participated in a large national cross-sectional health survey. Body mass index, smoking index and age were used as covariates in all analyses. RESULTS: Among the study population, 89.8% reported varying levels of coffee consumption; 6.9% were abstainers from alcohol, 86.1% moderate drinkers, 3.7% heavy drinkers and 3.3% former drinkers. In men, the elevation of GGT induced by heavy drinking (>280 g/week) was found to be significantly reduced by coffee consumption exceeding 4 cups per day. A similar trend was also observed among women, which however, did not reach statistical significance. CONCLUSION: Coffee modulates the effect of ethanol on serum GGT activities in a dose- and gender-dependent manner. These observations should be implicated in studies on the possible hepatoprotective effects of coffee in alcohol consumers.

Immunoassay for ethyl glucuronide in vitreous humor: a new tool for postmortem diagnostics of alcohol use.

Although excessive alcohol consumption plays a major role in fatal events, the role of alcohol use as a possible contributing factor at the time of death is not easy to establish due to lack of suitable biomarkers for postmortem analyses. We used an immunological approach to measure ethyl glucuronide (EtG) concentrations from vitreous humor (VH) and serum from 58 individuals representing a forensic autopsy population of cases with either a well-documented history of excessive alcohol use (n=37) or cases without such history (n=21), according to medical and police records and blood alcohol determinations (BAC). The immunoassay was based on the Microgenics DRI-EtG EIA reagents applied on an automated Abbott Architect c8000 clinical chemistry analyzer. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) determination of EtG and ethyl sulfate (EtS) was used as a reference method. At a cut-off of 0.3mg/l for VH-EtG, the immunoassay correctly identified 92% of the cases with a history of excessive alcohol use, whereas the BAC was positive (cut-off 10mg/dl) in 68% of the cases. A significant correlation emerged between VH-EtG and serum EtG (r=0.77, p<0.001) and between VH-EtG and BAC (r=0.62, p<0.001), although VH-EtG was frequently elevated also in cases with no detectable BAC. The EtG immunoassay showed a strong correlation with the LC-MS/MS reference method (r=0.94, p<0.001) and there was 100% agreement in the frequency of marker positive and negative findings between the immunoassay EtG results and the LC-MS/MS analysis of EtG and EtS. The present data indicate that the immunoassay for VH-EtG is a useful forensic tool for screening of antemortem alcohol use.

Effect of age and gender on the relationship between alcohol consumption and serum GGT: time to recalibrate goals for normal ranges.

AIMS: While serum gamma-glutamyltransferase (GGT) enzyme activity is a well established biomarker of excessive alcohol consumption and liver dysfunction, recent studies have also implicated it as a predictor of morbidity due to extrahepatic causes. Therefore, further information on the associations between ethanol intake and GGT activities in apparently healthy individuals appears warranted. METHODS: Data on alcohol consumption and serum GGT activities were collected from 18,899 individuals (8807 men, 10,092 women), mean age 48 years and range 25-74 years, who participated in a national cross-sectional health survey. Alcohol use was assessed by detailed questionnaires and the study population was subsequently divided into subgroups according to age and gender. Body mass index and smoking were used as covariates in all analyses. RESULTS: In men over 40 years, a reported regular consumption of 8 standard ethanol doses ('dose' = 12 g ethanol) or more per week was found to lead to a significant elevation in serum GGT activities, whereas those below 40 showed first significant changes not until the reported ethanol intake exceeded 14 doses per week. For women, the corresponding threshold levels were four and seven standard ethanol doses, respectively. CONCLUSION: The data pertaining to the present population sample indicate that rather low levels of reported regular ethanol consumption lead to elevated levels of GGT and that age over 40 markedly enhances the impact of alcohol consumption on GGT activity. The present findings should form the basis for defining safe levels of ethanol consumption and in recalibrating goals for normal limits in the clinical use of GGT measurements.