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.     

Genetic and environmental influences underlying externalizing behaviors, cigarette smoking and illicit drug use across adolescence

We investigated genetic and environmental influences common to adolescent externalizing behavior (at age 12), smoking (at age 14) and initiation of drug use (at age 17) using the FinnTwin12 cohort data. Multivariate Cholesky models were fit to data from 737 monozygotic and 722 dizygotic twin pairs. Heritability of externalizing behavior was 56%, that of smoking initiation/amount 20/32%, and initiation of drug use 27%. In the best-fitting model common environmental influences explained most of the covariance between externalizing behavior and smoking initiation (69%) and amount (77%). Covariance between smoking initiation/amount and drug use was due to additive genetic (42/22%) and common environmental (58/78%) influences. Half of the covariance between externalizing behavior and drug use was due to shared genetic and half due to the environments shared by co-twins. Using a longitudinal, prospective design, our results indicate that early observed externalizing behavior provides significant underlying genetic and environmental influences common to later substance use, here manifested as initiation of drug use in late adolescence.     

Vegetarianism and Eating-Disordered Thinking

Abstract

Two studies compared food choice motives and symptoms of eating disorders among vegetarians and nonvegetarians. The participants filled in a food choice questionnaire and completed an eating attitude test (EAT, Study 1) and eating disorder inventory (EDI, Study 2). The vegetarians scored higher on EAT and on the ineffectiveness, interpersonal distrust, and maturity fears suhscales of EDI than the nonvegetarians. However, no difference was found in the reported importance of weight control among the two groups. The results indicate that vegetarianism and eating disorders are not independent but rather are intertwined phenomena. The potential common links, for example the possibility that vegetarianism is being used as a smokescreen for more severe eating pathology, are discussed.     

Dynamin inhibition interferes with inflammasome activation and cytokine gene expression in Streptococcus pyogenes-infected human macrophages

In the present study, we have analysed the ability of Streptococcus pyogenes [Group A streptococcus (GAS)] to activate the NACHT-domain-, leucine-rich repeat- and PYD-containing protein 3 (NALP3) inflammasome complex in human monocyte-derived macrophages and the molecules and signalling pathways involved in GAS-induced inflammatory responses. We focused upon analysing the impact of dynamin-dependent endocytosis and the role of major streptococcal virulence factors streptolysin O (SLO) and streptolysin S (SLS) in the immune responses induced by GAS. These virulence factors are involved in immune evasion by forming pores in host cell membranes, and aid the bacteria to escape from the endosome–lysosome pathway. We analysed cytokine gene expression in human primary macrophages after stimulation with live or inactivated wild-type GAS as well as with live SLO and SLS defective bacteria. Interleukin (IL)-1β, IL-10, tumour necrosis factor (TNF)-α and chemokine (C-X-C motif) ligand (CXCL)-10 cytokines were produced after bacterial stimulation in a dose-dependent manner and no differences in cytokine levels were seen between live, inactivated or mutant bacteria. These data suggest that streptolysins or other secreted bacterial products are not required for the inflammatory responses induced by GAS. Our data indicate that inhibition of dynamin-dependent endocytosis in macrophages attenuates the induction of IL-1β, TNF-α, interferon (IFN)-β and CXCL-10 mRNAs. We also observed that pro-IL-1β protein was expressed and efficiently cleaved into mature-IL-1β via inflammasome activation after bacterial stimulation. Furthermore, we demonstrate that multiple signalling pathways are involved in GAS-stimulated inflammatory responses in human macrophages.     

Isokinetic thigh muscle performance after long-term recovery from patellar dislocation

Eighty-two patients (50 women, 32 men) underwent isokinetic muscle testing on average 13 years after a conservatively treated unilateral primary patellar dislocation. Three study groups were formed according to the natural history of recovery: group A (n = 32), patients with only primary conservative treatment; group B (n = 34) patients with conservative (group B1; ¶n = 24) or surgical (group B2; n = 10) treatment of redislocations; group C (n = 16) patients with other residual complaints (anterior knee, pain subluxations) requiring surgery. The Cybex 6000 dynamometer system was used as the testing machine for quadriceps and hamstrings muscles, with proportional deficits of peak torque as the test parameter. Isokinetic testing revealed both quadriceps and hamstring muscle atrophy even after long-term recovery from injury. There were statistically significant differences between the three study groups at both tested speeds of quadriceps muscles ¶(60 rad/s, P < 0.002; 180 rad/s, ¶P < 0.009). Groups B1 and B2 presented similar results. The muscle performance findings are probably due to more than one factor: primary immobilization, poor outcome, patellofemoral degeneration, redislocations, and residual knee complaints followed by surgery and deficiency in motor control of thigh muscle had – together or separately – an effect on muscle performance.     

Alveolar soft part sarcoma: a report of two cases with some histochemical and ultrastructural observations.

In a survey of all malignant soft tissue tumors in the extremities and limb girdles in Finland between 1960 and 1969, only one alveolar soft part sarcoma was found among 246 tumors (0.4%). Another alveolar soft part sarcoma, diagnosed in 1976, was more thoroughly studied. There was evidence that the characteristic crystals of alveolar soft part sarcoma are formed from the dense granules. Both were PASM-positive at ultrastructural level. No monoamines were detected in the cells by formaldehyde-induced fluorescence. This is a further fact to nullify the theory of the paraganglionic origin of alveolar soft part sarcoma, but the question of the histogenesis of the tumor still remains open.     

Vegetarianism and Eating-Disordered Thinking

Two studies compared food choice motives and symptoms of eating disorders among vegetarians and nonvegetarians. The participants filled in a food choice questionnaire and completed an eating attitude test (EAT, Study 1) and eating disorder inventory (EDI, Study 2). The vegetarians scored higher on EAT and on the ineffectiveness, interpersonal distrust, and maturity fears suhscales of EDI than the nonvegetarians. However, no difference was found in the reported importance of weight control among the two groups. The results indicate that vegetarianism and eating disorders are not independent but rather are intertwined phenomena. The potential common links, for example the possibility that vegetarianism is being used as a smokescreen for more severe eating pathology, are discussed.     

Live Lactobacillus rhamnosus and Streptococcus pyogenes differentially regulate Toll-like receptor (TLR) gene expression in human primary macrophages

Macrophages are phagocytes that recognize bacteria and subsequently activate appropriate innate and adaptive immune responses. TLRs are essential in identifying conserved bacterial structures and in initiating and mediating innate immune responses. In this work, we have characterized TLR gene expression in human monocyte-derived macrophages in response to stimulation with two live Gram-positive bacteria, a human commensal and probiotic Lactobacillus rhamnosus GG (LGG), and an important human pathogen Streptococcus pyogenes. LGG and S. pyogenes enhanced TLR2 expression in macrophages. LGG and S. pyogenes also required TLR2 for NF-kappaB activation. Only pathogenic S. pyogenes was able to up-regulate TLR3 and TLR7 gene expression. This up-regulation was dependent on IFN-alpha/beta, as neutralizing anti-IFN-alpha/beta antibodies reduced S. pyogenes-induced TLR3 and TLR7 mRNA expression. Our results show that despite similarities, TLR responses of macrophages differ for a Gram-positive probiotic and a pathogen. Our data suggest that macrophages can discriminate between probiotic and pathogenic bacteria by IFN-mediated TLR gene regulation.     

Simple measurements in assessing muscle performance after an ACL reconstruction

The purpose of this study was to evaluate the validity of the simple measurements of the muscle performance of the lower extremity, one legged hop testing for distance and the measurement of the circumference of the thigh 15 cm proximal to the joint line, by comparing them to the isokinetic strength testing 5 to 9 years after an ACL reconstruction with a bone-patellar tendon-bone (BTB) autograft. The measurements were performed on 86 patients on average 7 years after the surgery. The clinical evaluation was performed using the standard knee ligament evaluation form of the International Knee Documentation Committee (IKDC) and the Lysholm and the Marshall knee scores. At seven years, the isokinetic mean strength deficit of knee extension at the 60 degrees per second was 10 % in the operated limb as compared to the contralateral limb (NS). In the knee flexion, the differences were even smaller and statistically not significant either. The strength deficit of the knee extension at all knee angle velocities (p < 0.005), and flexion at 60 degrees per second (p < 0.05), correlated to the one legged hop testing, so that the patients with the greatest strength deficit also had the worst outcome in the one legged hop test. Also, the correlation between thigh atrophy and the deficit in the isokinetic strength test was significant in knee extension at all knee angle velocities (p < 0.001), and in knee flexion at 180 degrees per second (p < 0.005). In the final evaluation of the IKDC, 21 patients were rated as "normal", 51 as "nearly normal", 13 as "abnormal", and one as "severely abnormal". The mean of the Lysholm score was 83, classified as good, and that of Marshall score 43, classified as good. In conclusion, the one legged hop testing, as a functional muscle strength testing after an anterior cruciate ligament reconstruction, seems to correlate well with the isokinetic strength testing of the knee, especially in the knee extension. Because the hop testing can be easily performed and without extra equipment, we recommend its use for the evaluation of the functional muscle performance after an anterior cruciate ligament reconstruction. The measurement of the thigh atrophy is also easy to perform, and should be used beside the one legged hop testing, especially if the isokinetic strength testing is not available.