Biomarkers of alcohol consumption in patients classified according to the degree of liver disease severity.

OBJECTIVE: In the search for optimal biomarkers of excessive drinking, only a few studies have been conducted to compare the relationships between ethanol consumption, liver status, and various laboratory markers of ethanol-induced diseases. MATERIAL AND METHODS: Concentrations of carbohydrate-deficient transferrin (%CDT and CDTect methods), serum sialic acid (SA), gamma-glutamyl transferase (gamma-GT), aspartate aminotransferase (ASAT), mean corpuscular volume (MCV), and a marker of fibrogenesis (PIIINP) were studied in 102 alcoholics with (n=59) or without (n=43) alcoholic liver disease. Controls were 34 healthy volunteers who were either social drinkers or abstainers. RESULTS: Although concentrations of all markers were significantly higher in the alcoholic patients than in the healthy controls, their diagnostic characteristics showed a considerable degree of variation. The %CDT, SA, and MCV showed the strongest correlations with the amount of recent alcohol intake. The presence of liver pathology notably influenced the results of CDTect, GT, ASAT, and PIIINP. In ROC analyses, the highest rates of diagnostic accuracy for detecting hazardous drinking were reached with GT (0.94), CDT (0.86), and SA (0.85), followed by MCV (0.79) and ASAT (0.77). Upon abstinence, the estimated times for normalization varied between 10 days (CDTect) and 25 days (GT). CONCLUSIONS: Our data suggest distinct differences in the clinical characteristics of biological markers of ethanol consumption. While the overall accuracy of CDT and GT appear to be highest in the detection of problem drinking, serum SA and PIIINP measurements are of further value when the effects of liver pathology and ethanol drinking need to be differentiated.

Assays for acetaldehyde-derived adducts in blood proteins based on antibodies against acetaldehyde/lipoprotein condensates.

BACKGROUND: Acetaldehyde-derived protein condensates (adducts) have been suggested as promising biological markers of alcohol abuse because they represent actual metabolites of ethanol. However, the detection of such condensates in vivo has been hampered by a lack of sensitive and specific methods. METHODS: To develop new approaches for the detection of acetaldehyde adducts, we have raised antibodies against condensates with acetaldehyde and lipoproteins, which have previously been shown to be readily modified by acetaldehyde in vitro. The characteristics of these antibodies were compared with those raised against bovine serum albumin/acetaldehyde adduct and against other types of lipoprotein modifications, as induced by malondialdehyde, oxidation, and acetylation. RESULTS: The antibodies raised against low-density lipoprotein (LDL)/acetaldehyde, very low density lipoprotein (VLDL)/acetaldehyde, and bovine serum albumin/acetaldehyde all reacted with protein adducts generated at physiologically relevant concentrations of acetaldehyde in vitro, whereas the antibodies raised against malondialdehyde/LDL, oxidized LDL, or acetylated LDL were not found to cross-react with the acetaldehyde-derived adducts. In assays for acetaldehyde adducts from erythrocyte and serum proteins of patients with excessive ethanol consumption (n = 32) and healthy control individuals (n = 22), the antibody prepared against the acetaldehyde/VLDL condensate was found to provide the most effective detection of acetaldehyde adducts in vivo. CONCLUSIONS: Current data indicate that acetaldehyde generates immunogenic adducts with lipoproteins in vivo. Antibodies raised against the VLDL/acetaldehyde may provide a basis for new diagnostic assays to examine excessive alcohol consumption.

Acetaldehyde adducts in blood and bone marrow of patients with ethanol-induced erythrocyte abnormalities.

BACKGROUND: Although alcohol abuse is known to cause a wide array of adverse effects on blood cell formation, the molecular mechanisms by which alcohol exerts its toxic actions remain poorly defined. We examine here the formation of acetaldehyde-derived protein modifications in erythrocytes and in their bone marrow precursors using antibodies specifically recognizing acetaldehyde-modified epitopes in proteins independently of the nature of the carrier protein. MATERIALS AND METHODS: We studied 138 consecutive adult patients undergoing bone marrow aspiration due to macrocytosis (MCV values above 99 fL). Assessment included complete blood counts, morphologic review, assessment of alcohol consumption, and biochemical and immunocytochemical assays for acetaldehyde adducts. RESULTS: There were 68 patients (49%) with a history of excessive alcohol consumption, 28 (20%) of whom were patients with severe dependence. The blood smears prepared from the alcoholic patients with macrocytosis also contained stomatocytes and knizocytes. Bone marrow aspirates from 12 alcoholic patients showed vacuolization of pronormoblasts and the presence of ring sideroblasts was noted in 8 cases. In immunocytochemical analyses of the peripheral blood erythrocytes, acetaldehyde-derived epitopes were found to occur both on the cell membrane and inside the erythrocytes. Bone marrow aspirates also showed positive staining for acetaldehyde adducts in the erythropoietic cells in 8 of 11 (73%) consecutive alcoholic patients. Separation of the erythrocyte proteins from the samples of alcoholics on HPLC-chromatography revealed the formation of fast-eluting hemoglobin fractions, which also reacted with antibodies against acetaldehyde adducts. CONCLUSIONS: Current data suggest that acetaldehyde-erythrocyte adducts are formed in vivo in blood and bone marrow of patients with excessive alcohol consumption. This may contribute to the generation of the erythrocyte abnormalities, which are frequently observed in alcoholic patients.

Cell-mediated immune response during primary Chlamydia pneumoniae infection.

The development of Chlamydia pneumoniae-specific cell-mediated immunity was studied during a primary C. pneumoniae infection. The immune response was detected as positive lymphocyte proliferation and secretion of interferon gamma. C. pneumoniae-induced activation of both CD4(+) and CD8(+) T cells was detected in the early phase of infection, but activation of only CD4(+) T cells was detected in the later stage.

Lysyl hydroxylase 3 is a multifunctional protein possessing collagen glucosyltransferase activity.

Lysyl hydroxylase (EC ) and glucosyltransferase (EC ) are enzymes involved in post-translational modifications during collagen biosynthesis. We reveal in this paper that the protein produced by the cDNA for human lysyl hydroxylase isoform 3 (LH3) has both lysyl hydroxylase and glucosyltransferase (GGT) activities. The other known lysyl hydroxylase isoforms, LH1, LH2a, and LH2b, have no GGT activity. Furthermore, antibodies recognizing the amino acid sequence of human LH3 and those against a highly purified chicken GGT partially inhibited the GGT activity. Similarly, a partial inhibition was observed when these antibodies were tested against GGT extracted from human skin fibroblasts. In vitro mutagenesis experiments demonstrate that the amino acids involved in the GGT active site differ from those required for LH3 activity.

Cold-induced responses in the upper respiratory tract.

Recent investigations have shown that the prevalence of bronchial asthma is higher among skiers exposed to cold and dry air than among nonskiers. The upper airway passages are responsible for warming and humidifying the inhaled air. During exercise in cold and dry air, warming and humidifying of the inhaled air continues in the bronchial tree. Under these conditions both nasal and bronchial mucosa are cooled by inspiratory air and remain cooled throughout the respiratory cycle. The air which reaches the bronchoalveolar air space is at body temperature and fully saturated in all conditions. In this article we briefly review the studies carried out regarding the effects of cold air inhalation on the upper respiratory tract.

Water balance and physical performance in cold.

Although a lot of attention is focused on problems of energy balance and nutrition in cold environment, water balance has received less attention. In a cold environment the water balance might be disturbed because the need of water could be increased, the use of water could be decreased and the redistribution of blood could change water volume in circulation. In dehydrated subjects, exercising at -15 degrees C at submaximal work level, both oxygen uptake and heart rate were significantly higher during water deprivation while the anaerobic threshold was lower. The time before reaching exhaustion was also shortened. However, maximal oxygen uptake and maximal muscle strength were not affected. The results suggest lower efficiency, higher physical strain and earlier exhaustion of dehydrated subjects in cold. After repletion of 1.8% body weight loss by an equal amount of fluid (5% sucrose solution) the oxygen uptake was significantly decreased at a submaximal work level at -20 degrees C, suggesting an improved mechanical efficiency and decreased physical strain. Although physical performance could be restored by rehydration, a rapid rehydration is not recommended because of increased diuresis, increased blood pressure and vigorous shivering stimulated by cold fluids. Instead, a continuous maintenance of water balance is recommended, with a fluid temperature above 25-30 degrees C and with a carbohydrate content below 7%.

Sauna builder's asthma caused by obeche (Triplochiton scleroxylon) dust.

Two carpenters developed rhinitis, conjunctivitis, bronchial wheezing, and dyspnoea while using obeche (Triplochiton scleroxylon, African maple) at their work while they built saunas. Skin prick tests showed an immediate reaction, specific IgE to obeche was detected in their serum, and bronchial provocation test with obeche gave an immediate reaction with decrease of FEV1 and PEF values. The symptoms disappeared after avoiding the use of obeche. Obeche may cause a health hazard to carpenters who are exposed to this dust and who may develop allergic symptoms after the exposure.