31.3.2008

Alanne, M., Kristiansson, K., Auro, K., Silander, K., Kuulasmaa, K., Peltonen, L., Salomaa, V. and Perola, M.

Hum Genet. 2007; 122(3-4): 355-65. IF 3.662

Selenoprotein S (SEPS1) is a novel candidate gene involved in the regulation of inflammatory response and protection from oxidative damage.

This study explored the genetic variation in the SEPS1 locus for an association with CVD as well as with quantitative phenotypes related to obesity and inflammation. We used the case-cohort design and time-to-event analysis in two separate prospectively followed population-based cohorts FINRISK 92 and 97 (n = 999 and 1,223 individuals, respectively) to study the associations of five single nucleotide polymorphisms with the risk for coronary heart disease (CHD) and ischemic stroke events. We found a significant association with increased CHD risk in females carrying the minor allele of rs8025174 in the combined analysis of both cohorts [hazard ratio (HR) 2.95 (95% confidence interval: 1.37-6.39)]. Another variant, rs7178239, increased the risk for ischemic stroke significantly in females [HR: 3.35 (1.66-6.76)] and in joint analysis of both sexes and both cohorts [HR: 1.75 (1.17-2.64)].

These results indicate that variation in the SEPS1 locus may have an effect on CVD morbidity, especially in females. This observation should stimulate further investigations of the role of this gene and protein in the pathogenesis of CVD.

Treatment of Hypertension in Patients 80 Years of Age or Older

Beckett, N. S., Peters, R., Fletcher, A. E., Staessen, J. A., Liu, L., Dumitrascu, D., Stoyanovsky, V., Antikainen, R. L., Nikitin, Y., Anderson, C., Belhani, A., Forette, F., Rajkumar, C., Thijs, L., Banya, W. and Bulpitt, C. J.

N Engl J Med. 2008. IF 51.296

BACKGROUND: Whether the treatment of patients with hypertension who are 80 years of age or older is beneficial is unclear. It has been suggested that antihypertensive therapy may reduce the risk of stroke, despite possibly increasing the risk of death.

METHODS: We randomly assigned 3845 patients from Europe, China, Australasia, and Tunisia who were 80 years of age or older and had a sustained systolic blood pressure of 160 mm Hg or more to receive either the diuretic indapamide (sustained release, 1.5 mg) or matching placebo. The angiotensin-converting-enzyme inhibitor perindopril (2 or 4 mg), or matching placebo, was added if necessary to achieve the target blood pressure of 150/80 mm Hg. The primary end point was fatal or nonfatal stroke.

RESULTS: The active-treatment group (1933 patients) and the placebo group (1912 patients) were well matched (mean age, 83.6 years; mean blood pressure while sitting, 173.0/90.8 mm Hg); 11.8% had a history of cardiovascular disease. Median follow-up was 1.8 years. At 2 years, the mean blood pressure while sitting was 15.0/6.1 mm Hg lower in the active-treatment group than in the placebo group. In an intention-to-treat analysis, active treatment was associated with a 30% reduction in the rate of fatal or nonfatal stroke (95% confidence interval [CI], -1 to 51; P=0.06), a 39% reduction in the rate of death from stroke (95% CI, 1 to 62; P=0.05), a 21% reduction in the rate of death from any cause (95% CI, 4 to 35; P=0.02), a 23% reduction in the rate of death from cardiovascular causes (95% CI, -1 to 40; P=0.06), and a 64% reduction in the rate of heart failure (95% CI, 42 to 78; P<0.001). Fewer serious adverse events were reported in the active-treatment group (358, vs. 448 in the placebo group; P=0.001).

CONCLUSIONS: The results provide evidence that antihypertensive treatment with indapamide (sustained release), with or without perindopril, in persons 80 years of age or older is beneficial. (ClinicalTrials.gov number, NCT00122811.) Copyright 2008 Massachusetts Medical Society.

Within-family outliers: segregating alleles or environmental effects? A linkage analysis of height from 5815 sibling pairs

Benyamin, B., Perola, M., Cornes, B. K., Madden, P. A. F., Palotie, A., Nyholt, D. R., Montgomery, G. W., Peltonen, L., Martin, N. G. and Visscher, P. M.

European Journal of Human Genetics. 2008; 16(4): 516-524. Article. IF 3.697

Most information in linkage analysis for quantitative traits comes from pairs of relatives that are phenotypically most discordant or concordant. Confounding this, within-family outliers from non-genetic causes may create false positives and negatives. We investigated the influence of within-family outliers empirically, using one of the largest genome-wide linkage scans for height. The subjects were drawn from Australian twin cohorts consisting of 8447 individuals in 2861 families, providing a total of 5815 possible pairs of siblings in sibships. A variance component linkage analysis was performed, either including or excluding the within-family outliers. Using the entire dataset, the largest LOD scores were on chromosome 15q (LOD 2.3) and 11q (1.5). Excluding within-family outliers increased the LOD score for most regions, but the LOD score on chromosome 15 decreased from 2.3 to 1.2, suggesting that the outliers may create false negatives and false positives, although rare alleles of large effect may also be an explanation. Several regions suggestive of linkage to height were found after removing the outliers, including 1q23.1 (2.0), 3q22.1 (1.9) and 5q32 (2.3). We conclude that the investigation of the effect of within-family outliers, which is usually neglected, should be a standard quality control measure in linkage analysis for complex traits and may reduce the noise for the search of common variants of modest effect size as well as help identify rare variants of large effect and clinical significance. We suggest that the effect of within-family outliers deserves further investigation via theoretical and simulation studies.

Coffee consumption and type 2 diabetes - An extensive review

Bidel, S., Hu, G. and Tuomilehto, J.

Central European Journal of Medicine. 2008; 3(1): 9-19. Review.

Coffee is a complex mixture of potentially active chemicals. It contains significant amounts of phenolic polymers, chlorogenic acid and also caffeine. Agricultural factors, roasting, blending, and brewing determine coffee's chemical composition. Recent epidemiological studies suggest that habitual coffee consumption may help to prevent some chronic diseases including type 2 diabetes. Despite reports from the clinical trials of the effect of caffeine on decreasing insulin sensitivity, long-term prospective studies revealed that coffee may improve fasting glucose, glucose tolerance and insulin sensitivity as well. In the most recent publication habitual coffee drinkers have a lower total and cardiovascular mortality rate among diabetic subjects.

Modulation of voluntary ethanol consumption by beta-arrestin 2

Bjork, K., Rimondini, R., Hansson, A. C., Terasmaa, A., Hyytiä, P., Heilig, M. and Sommer, W. H.

FASEB J. 2008. IF 6.721

Beta-arrestin 2 is a multifunctional key component of the G protein-coupled receptor complex and is involved in micro-opiate and dopamine D2 receptor signaling, both of which are thought to mediate the rewarding effects of ethanol consumption. We identified elevated expression of the beta-arrestin 2 gene (Arrb2) in the striatum and the hippocampus of ethanol-preferring AA rats compared to their nonpreferring counterpart ANA line. Differential mRNA expression was accompanied by different levels of Arrb2 protein. The elevated expression was associated with a 7-marker haplotype in complete linkage disequilibrium, which segregated fully between the lines, and was unique to the preferring line. Furthermore, a single, distinct, and highly significant quantitative trait locus for Arrb2 expression in hippocampus and striatum was identified at the locus of this gene, providing evidence that genetic variation may affect a cis-regulatory mechanism for expression and regional control of Arrb2. These findings were functionally validated using mice lacking Arrb2, which displayed both reduced voluntary ethanol consumption and ethanol-induced psychomotor stimulation. Our results demonstrate that beta-arrestin 2 modulates acute responses to ethanol and is an important mediator of ethanol reward.-Bjork, K., Rimondini, R., Hansson, A. C., Terasmaa, A., Hyytia, P., Heilig, M., Sommer, W. H. Modulation of voluntary ethanol consumption by beta-arrestin 2.

Flavonoid intake and risk of pancreatic cancer in male smokers (Finland)

Bobe, G., Weinstein, S. J., Albanes, D., Hirvonen, T., Ashby, J., Taylor, P. R., Virtamo, J. and Stolzenberg-Solomoni, R. Z.

Cancer Epidemiology Biomarkers & Prevention. 2008; 17(3): 553-562. Article. IF 4.289

Extending research on the protective effect of flavonoids in cell culture and animal studies, we examined the association between consumption of flavonoids and flavonoid-rich foods and development of exocrine pancreatic cancer within the alpha-Tocopherol, beta-Carotene Cancer Prevention Study cohort. Of the 27,111 healthy male smokers (50-69 years) who completed a self-administered dietary questionnaire at baseline, 306 developed exocrine pancreatic cancer during follow-up (1985-2004; median, 16.1 years). Intakes of total flavonoids, three flavonoid subgroups, seven individual flavonoids, and flavonoid-rich foods were estimated from a validated food frequency questionnaire. Hazard ratios and 95% confidence intervals were estimated using Cox proportional hazards models. Overall, flavonoid intake was not significantly associated with pancreatic cancer. However, in stratified analysis, greater total flavonoid intake was associated with decreased pancreatic cancer risk in participants randomized during the trial to placebo (fourth versus first quartile: hazard ratio, 0.36; 95% confidence interval, 0.17-0.78; P-trend = 0.009) and not to supplemental alpha-tocopherol (50 mg/d) and/or beta-carotene (20 mg/d; P-interaction = 0.002). Similar patterns and significant interactions were observed for flavonols, flavan-3-ols, kaempferol, quercetin, catechin, and epicatechin. Our data suggest that a flavonoid-rich diet may decrease pancreatic cancer risk in male smokers not consuming supplemental alpha-tocopherol and/or beta-carotene.

The physical strenuousness of work is slightly associated with an upward trend in the BMI

Bockerman, P., Johansson, E., Jousilahti, P. and Uutela, A.

Social Science & Medicine. 2008; 66(6): 1346-1355. Article. IF 2.749

This paper examines the relationship between the physical strenuousness of work and the BMI in Finland, using individual microdata at 5-year intervals over the period 1972-2002. Data came from the National FINRISK Study which contains self-reported information on the physical strenuousness of a respondent's occupation. Our estimates show that the changes in the physical strenuousness of work explain around 7% at most of the increase in BMI for Finnish males observed over a period of 30 years. The main reason for this appears to be the effect of the physical strenuousness of work on BMI which is rather moderate. According to the point estimates, BMI is 2.4% lower when a male's occupation is physically very demanding and involves lifting and carrying heavy objects compared with a sedentary job (reference group of the estimations), other things being equal. Furthermore, it is very difficult to associate the changes in the occupational structure with the upward trend in BMI for females, and the contribution of the changes in the occupational structure is definitely even smaller for females than it is for males. All in all, we show that the changes in self-reported occupation show a slight association with the changes in the logarithm of the BMI scores. (c) 2007 Elsevier Ltd. All rights reserved.

Association of age and education with different types of leisure-time physical activity among 4437 finnish adults

Borodulin, K., Laatikainen, T., Lahti-Koski, M., Jousilahti, P. and Lakka, T. A.

J Phys Act Health. 2008; 5(2): 242-51.

Background: The aim of this study was to investigate the associations of age and education with types of leisure-time physical activity in a population sample of Finnish adults. Methods: The sample, part of the National FINRISK Study, comprised 1940 men and 2497 women age 25 to 64 years. Self-reported questionnaires were used to collect data on education and leisure-time physical activity. Overall leisure-time physical activity was further divided into conditioning and daily physical activity. Results: In men and women, age had an inverse association with conditioning physical activity but not with daily and overall leisure activity. Strong direct associations were found between education and conditioning and overall leisure activity. Conclusions: All age groups report fairly similar levels of overall and daily levels of leisure-time physical activity, but the levels differ across educational groups. In health promotion, more emphasis should be targeted to the population groups with lower education.

Persistence of measles, mumps, and rubella antibodies in an MMR-vaccinated cohort: A 20-year follow-up

Davidkin, I., Jokinen, S., Broman, M., Leinikki, P. and Peltola, H.

Journal of Infectious Diseases. 2008; 197(7): 950-956. Article. IF 5.363

Background. The persistence of antibodies against measles, mumps, and rubella induced by the measles-mumps-rubella ( MMR) vaccine and the kinetics of antibody decline after the second MMR vaccine dose were studied in the same cohort for 20 years. Methods. Measles, mumps, and rubella antibodies were measured by enzyme immunoassay in 20-year follow-up serum samples ( n = 183) of twice-vaccinated individuals, and measles antibodies were also measured in oral fluids ( n = 177). Antibody decay was determined in a group ( n = 58) with subsequent samples collected 1, 8, and 15 years after the second MMR dose. Results. In total, 95%, 74%, and 100% of 183 vaccinees were still seropositive for measles, mumps, and rubella, respectively, and 85% of 177 vaccinees had measurable measles antibodies in their oral fluids. The antibody levels declined significantly after the second dose, but subsequently the rate of decline was slower. Conclusions. A high rate of seropositivity was found 20 years after the first MMR dose, particularly for rubella and measles. Our results show that MMR vaccine-induced antibodies wane significantly after the second dose. According to epidemiological data, the protection induced by MMR vaccination in Finland seems to persist at least until early adulthood. However, the situation requires constant vigilance.

Nuclear translocation of hypoxia-inducible factors (HIFs): Involvement of the classical importin alpha/beta pathway

Depping, R., Steinhoff, A., Schindler, S. G., Friedrich, B., Fagerlund, R., Metzen, E., Hartmann, E. and Kohler, M.

Biochimica Et Biophysica Acta-Molecular Cell Research. 2008; 1783(3): 394-404. Article. IF 6.900

Hypoxia-inducible factors are the key elements in the essential process of oxygen homeostasis of vertebrate cells. Stabilisation and subsequent nuclear localisation of HIF-alpha subunits results in the activation of target genes such as vegf, epo and glut1. The passage of transcription factors e.g. HIF-1 alpha into the nucleus through the nuclear pore complex is regulated by nuclear transport receptors. Therefore nucleocytoplasmic shuttling can regulate transcriptional activity by facilitating the cellular traffic of transcription factors between both compartments. Here, we report on the identification of specific interactions of hypoxia-inducible factors with nuclear transport receptors importin alpha/beta. HIF-1 alpha, -1 beta, and HIF-2 alpha are binding to importin alpha 1, alpha 3, alpha 5, and alpha 7. The direct interaction of HIF-1 alpha to alpha importins is dependent on a functional nuclear localisation signal within the C-terminal region of the protein. In contrast, the supposed N-terminal NLS is not effective. Our findings provide new insight into the mechanism of the regulation of nuclear transport of hypoxia-inducible factors. (c) 2007 Elsevier B.V. All rights reserved.

Preface to the proceedings of the satellite symposium of the EAS 76th Congress and the XVth Paavo Nurmi Symposium

Ehnholm, C., Jauhiainen, M., Olkkonen, V., Ikonen, E. and Kovanen, P.

Annals of Medicine. 2008; 40: 4-4. Editorial Material. IF 4.594

Favorable effects of berry consumption on platelet function, blood pressure, and HDL cholesterol

Erlund, I., Koli, R., Alfthan, G., Marniemi, J., Puukka, P., Mustonen, P., Mattila, P. and Jula, A.

Am J Clin Nutr. 2008; 87(2): 323-31. IF 6.562

BACKGROUND: Berries are a particularly rich source of polyphenols. They also contain other bioactive substances, such as vitamin C. Previous studies indicated that the consumption of polyphenol-rich foods (eg, cocoa, tea, and red wine) may induce beneficial changes in pathways related to cardiovascular health. Whether the consumption of berries has similar effects is unknown.

OBJECTIVE: We aimed to investigate the effects of berry consumption on hemostatic function, serum lipids, and blood pressure (BP).

DESIGN: Middle-aged unmedicated subjects (n = 72) with cardiovascular risk factors consumed moderate amounts of berry or control products for 8 wk in a single-blind, randomized, placebo-controlled intervention trial.

RESULTS: Berry consumption inhibited platelet function as measured with a platelet function analyzer (using collagen and ADP as platelet activator) [changes: 11% and -1.4% in the berry and control groups, respectively; P = 0.018, analysis of covariance (ANCOVA)]. Plasma biomarkers of platelet activation, coagulation, and fibrinolysis did not change during the intervention. Serum HDL-cholesterol concentrations increased significantly more (P = 0.006, ANCOVA) in the berry than in the control group (5.2% and 0.6%, respectively), but total cholesterol and triacylglycerol remained unchanged. Systolic BP decreased significantly (P = 0.050, ANCOVA); the decrease mostly occurred in subjects with high baseline BP (7.3 mm Hg in highest tertile; P = 0.024, ANCOVA). Polyphenol and vitamin C concentrations in plasma increased, whereas other nutritional biomarkers (ie, folate, tocopherols, sodium, and potassium) were unaffected.

CONCLUSION: The consumption of moderate amounts of berries resulted in favorable changes in platelet function, HDL cholesterol, and BP. The results indicate that regular consumption of berries may play a role in the prevention of cardiovascular disease.

Apolipoprotein E epsilon 4 allele is associated with increased atrophy in progressive mild cognitive impairment: A vaxel-based morphometric study

Hamalainen, A., Grau-Olivares, M., Tervo, S., Niskanen, E., Pennanen, C., Huuskonen, J., Kivipelto, M., Hanninen, T., Tapiola, M., Vanhanen, M., Hallikainen, M., Helkala, E., Nissinen, A., Vanninen, R. L. and Soininen, H.

Neurodegenerative Diseases. 2008; 5(3-4): 186-189. Article.

Background. The apolipoprotein E (APOE) epsilon 4 allele is a risk factor for Alzheimer's disease. Earlier studies have shown differences in brain structure according to the APOE 64 status. Objective: To assess possible differences in brain structure according to the APOE epsilon 4 status in mild cognitive impairment (MCI) subjects in relation to conversion to dementia. Methods: In a follow-up study of 56 MCI subjects, 13 MCI subjects progressed to dementia (PMCI) during a mean follow-up time of 31 months. Brain structure differences in both stable MCI (SMCI) and PMCl epsilon 4 carriers and noncarriers in the baseline MRI scan were assessed with voxel-based morphometry. Results: The SMCI epsilon 4 carriers had atrophy in the amygdala and hippocampus compared to the SMCI noncarriers. The PMCl epsilon 4 carriers revealed atrophy of the left inferior frontal gyrus and parietal cortex compared to the PMCI noncarriers. Conclusion: The rate of brain atrophy in certain brain areas may be increased in epsilon 4-positive MCI subjects progressing to dementia. Copyright (c) 2008 S. Karger AG, Basel.

Use of health services for major depressive and anxiety disorders in Finland

Hamalainen, J., Isometsa, E., Sihvo, S., Pirkola, S. and Kiviruusu, O.

Depress Anxiety. 2008; 25(1): 27-37. IF 2.549

Factors associated with people suffering from major depressive disorder (MDD) or anxiety disorders seeking or receiving treatment are not well known. In the Health 2000 Study, a representative sample (n=6005) of Finland's general adult (> or =30 years) population was interviewed with the M-CIDI for mental disorders and health service use for mental problems during the last 12 months. Predictors for service use among those with DSM-IV MDD (n=298) or anxiety disorders (n=242) were assessed. Of subjects with MDD, anxiety disorders, or both, 34%, 36%, and 59% used health services, respectively. Greater severity and perceived disability, psychiatric comorbidity, and living alone predicted health care use for MDD subjects, and greater perceived disability, psychiatric comorbidity, younger age, and parent's psychiatric problems for anxiety disorder subjects. The use of specialist-level mental health services was predicted by psychiatric comorbidity, but not characteristics of the disorders per se. Perceived disability and comorbidity are factors influencing the use of mental health services by both anxiety disorder and MDD subjects. However, still only approximately one-half of those suffering from even severe and comorbid disorders use health services for them.

Long-term outcome of major depressive disorder in psychiatric patients is variable

Holma, K. M., Holma, I. A., Melartin, T. K., Rytsala, H. J. and Isometsa, E. T.

J Clin Psychiatry. 2008; 69(2): 196-205. IF 5.533

OBJECTIVE: The prevailing view of outcome of major depressive disorder (MDD), based on mostly inpatient cohorts sampled from tertiary centers, emphasizes chronicity and frequent recurrences. We investigated the long-term outcome of a regionally representative psychiatric MDD cohort comprising mainly outpatients.

METHOD: The Vantaa Depression Study included 163 patients with DSM-IV MDD (71.5% of those eligible) diagnosed using structured and semistructured interviews and followed up at 6 months, 18 months, and 5 years with a life chart between February 1, 1997, and April 30, 2004. The effects of comorbid disorders and other predictors on outcome were comprehensively investigated.

RESULTS: Over the 5-year follow-up, 98.8% of patients achieved a symptom state below major depressive episode (MDE) criteria, and 88.4% reached full remission, with the median time to full remission being 11.0 months. Nearly one third (29.3%) had no recurrences, whereas 30.0% experienced 1, 12.9% experienced 2, and 27.9% experienced 3 or more recurrences. Preceding dysthymic disorder (p = .028), cluster C personality disorder (p = .041), and longer MDE duration prior to entry (p = .011) were the most significant predictors of longer time in achieving full remission. Severity of MDD and comorbidity, especially social phobia, predicted probability of, shorter time to, and number of recurrences.

CONCLUSION: Previous literature on mostly inpatient MDD may have, by generalizing from patients with the most severe psychopathology, overemphasized chronicity of MDD. The long-term outcome of MDD in psychiatric care is variable, with about one tenth of patients having poor, one third having intermediate, and one half having favorable outcomes. In addition to known predictors, cluster C personality disorders and social phobia warrant further attention as predictors of MDD outcome among outpatients.

Physical activity, physical fitness, and risk of type 2 diabetes mellitus

Hu, G., Lakka, T. A., Barengo, N. C. and Tuomilehto, J.

Metab Syndr Relat Disord. 2005; 3(1): 35-44.

Type 2 diabetes mellitus (DM) is a common chronic disease. Cardiovascular disease is the most prevalent complication of DM. In the past decade, the associations of physical activity, physical fitness, and changes in lifestyle with the risk of type 2 DM have been assessed by a number of prospective epidemiologic studies and clinical trials. Several studies also evaluate the joint associations of physical activity, body mass index, and glucose levels with the risk of type 2 DM. The results of 21 prospective studies and four clinical trials demonstrated that moderate or high levels of physical activity or physical fitness, as well as changes in lifestyle (dietary modification and enhanced physical activity) could prevent type 2 DM.

Lifestyle management in the metabolic syndrome

Hu, G., Lakka, T. A., Lakka, H. M. and Tuomilehto, J.

Metab Syndr Relat Disord. 2006; 4(4): 270-86.

Type 2 diabetes and metabolic syndrome are two of the fastest growing public health problems in both developed and developing countries. Cardiovascular disease is the most prevalent complication of type 2 diabetes and the metabolic syndrome. Overweight, obesity, or weight gain has been shown to be an important risk factor for the development of type 2 diabetes and an important component of the metabolic syndrome. Physical inactivity is another important risk factor for the development of type 2 diabetes. Data from prospective studies have shown that at least 30 min/day of moderate to vigorous physical activity can prevent type 2 diabetes. Moderate or high levels of physical fitness are effective in preventing type 2 diabetes. Results from clinical trials have indicated that lifestyle changes, including dietary modification and increase in physical activity, can prevent type 2 diabetes. Analyses from prospective studies have confirmed that healthy diets are effective and safe ways to prevent type 2 diabetes and the metabolic syndrome. Public health messages, health care professionals, and the health care system should aggressively promote physical activity and responsible nutritional habits during occupation, leisure time, and daily life and prevent overweight and obesity.

Functional interaction of AIRE with PIAS1 in transcriptional regulation

Ilmarinen, T., Kangas, H., Kytomaa, T., Eskelin, P., Saharinen, J., Seeler, J. S., Tanhuanpaa, K., Chan, F. Y. L., Slattery, R. M., Alakurtti, K., Palvimo, J. J. and Ulmanen, I.

Molecular Immunology. 2008; 45(7): 1847-1862. Article. IF 4.768

AIRE (autoimmune regulator) promotes the establishment of self-tolerance by regulating gene expression in the thymus. Mutations in AIRE lead to an autoimmune disease, APECED. Here we have identified PIAS proteins as novel AIRE interaction partners. Although PIAS proteins function as E3 SUMO ligases, AIRE is not sumoylated. We expressed AIRE, wt PIAS1, and PIAS1 mutants with deleted SP-RING domain or SUMO interaction motif (SIM) in different cell lines and demonstrate that AIRE and PIAS1 localize to adjacent nuclear bodies (NBs). The expression of AIRE enhances the formation of PIAS1 NBs. The ability of PIAS1 to localize into NBs and interconnect with AIRE is neither dependent on the SP-RING domain nor the SIM. Further, we show that PIAS1 is able to attract AIRE into SUMO1-containing complexes and that the process is dependent on the SIM of PIAS1. PIASI and AIRE concurrently activate the human insulin promoter, a known target gene of AIRE, and the SP-RING is required for this activation. Moreover, AIRE represses and PIAS1 activates the CSTB promoter, used as a model for a housekeeping promoter, and both the SP-RING and SIM are needed for its activation by PIAS1. Collectively, our data suggest that AIRE and PIAS1 interact functionally to regulate the activities of the target genes of AIRE. (C) 2007 Elsevier Ltd. All rights reserved.

Human herpesvirus-6 and-7 DNA in cerebrospinal fluid of facial palsy patients

Kanerva, M., Jaaskelainen, A. J., Suvela, M., Piiparinen, H., Vaheri, A. and Pitkaranta, A.

Acta Oto-Laryngologica. 2008; 128(4): 460-464. Article. IF 0.738

Conclusions. Finding human herpesvirus (HHV)-7 and dual HHV-6A and -6B DNA in cerebrospinal fluid (CSF) of two facial palsy (FP) patients is intriguing but does not allow etiologic conclusions as such. HHV-6 or -7 DNA was revealed in 10% of the CSF samples tested from 70 immunocompetent adolescents and adults; a highly unusual result. How these findings are associated with the diseases they accompany remains to be defined. Objective. To determine whether herpes simplex virus (HSV)-1 and -2, varicella-zoster virus (VZV), HHV-6A, -6B, and -7, Epstein-Barr virus (EBV), and cytomegalovirus (CMV) DNA could be found in CSF of FP patients or controls. Subjects and methods. In all, 33 peripheral FP patients (26 idiopathic, 5 with herpesvirus infection, 1 puerperal, 1 Melkersson-Rosenthal syndrome) (34 CSF samples) and 36 controls (16 nonidiopathic FP, 7 hearing loss, 6 vertigo, 5 headache, 2 other) previously tested for HSV-1, VZV, and HHV-6 DNA by polymerase chain reaction (PCR) were tested with highly sensitive multiplex-PCR and an oligonucleotide microarray method. Results. One FP patient had HHV-7 DNA and another had HHV-6A and -6B DNA simultaneously. In the control group, one HHV-7, one HHV-6A, and three HHV-6B DNA-positive specimens were found.

Childhood and Current Determinants of Heavy Drinking in Early Adulthood

Kestila, L., Martelin, T., Rahkonen, O., Joutsenniemi, K., Pirkola, S., Poikolainen, K. and Koskinen, S.

Alcohol Alcohol. 2008. IF 2.061

AIMS: To explore the association of parental education, childhood living conditions and several adversities with heavy drinking in early adulthood, and to analyze the effect of the respondent's current circumstances on these associations.

METHOD: The analyses were conducted in a sample of 1234 adults aged 18-29 years participating in the Finnish Health 2000 Survey (65% of the original representative two-stage cluster sample, N = 1894). The outcome measure was heavy drinking measured by g/week for pure alcohol (for men >/=280 g/week and for women >/=140 g/week).

RESULTS: 8% of young adult men and 5% of women were heavy drinkers. In both genders, parental alcohol problems and other childhood adversities, poor own education, and unemployment status increased the risk of heavy drinking. The impact of childhood on heavy drinking was partly independent and partly mediated by adult characteristics, in particular, for both genders, low level of education.

CONCLUSIONS: Childhood adversities are associated with heavy drinking in early adulthood among both genders. Childhood social circumstances as well as low educational level and unemployment should be taken into account in planning preventive policies to tackle the harms caused by excessive alcohol use at the individual and population level.

Coronary events in persons aged 75 years or older in Finland from 1995 to 2002: The FINAMI study

Koukkunen, H., Salomaa, V., Lehto, S., Ketonen, M., Immonen-Raiha, P., Lehtonen, A., Havulinna, A., Kesaniemi, Y. A. and Pyorala, K.

American Journal of Geriatric Cardiology. 2008; 17(2): 78-86. Article.

The authors used population-based myocardial infarction (MI) register data to examine trends in incidence, case fatality, treatment strategies of MI, and coronary heart disease (CHD) mortality in persons aged 75 to 99 years in 4 areas of Finland during 1995 through 2002. This age group contributed 53% (n=13,977) of all CHD events, and 65% occurred in women. CHD mortality declined among men annually by 3.5% and 1.0% in the 75- to 84-year-old and 85- to 99-year-old age groups, respectively. Among women, it declined by 2.2% per year in the 75- to 84-year-old age group but increased by 1.3% per year in the 85- to 99-year-old age group. MI attack rate did not change in men but increased significantly in women aged 85 to 99 years. Clinical management of MI in elderly patients was more conservative than in middle-aged patients. In conclusion, one-half of all CHD events occur among persons aged 75 years or older, and elderly patients with CHD represent an increasing burden to the health care system.

Transitions into and out of daylight saving time compromise sleep and the rest-activity cycles

Lahti, T. A., Leppamaki, S., Lonnqvist, J. and Partonen, T.

BMC Physiol. 2008; 8: 3.

BACKGROUND: The aim of this study was to analyze the effects of transition out of and into daylight saving time on the rest-activity cycles and sleep. Rest-activity cycles of nine healthy participants aged 20 to 40 years were measured around transitions out of and into daylight saving time on fall 2005 and spring 2006 respectively. Rest-activity cycles were measured using wrist-worn accelerometers. The participants filled in the Morningness-Eveningness and Seasonal Pattern Assessment Questionnaires before starting the study and kept a sleep diary during the study.

RESULTS: Fall transition was more disturbing for the more morning type and spring transition for the more evening type of persons. Individuals having a higher global seasonality score suffered more from the transitions.

CONCLUSION: Transitions out of and into daylight saving time enhanced night-time restlessness and thereby compromised the quality of sleep.

The effect of birth order and parental age on the risk of type 1 and 2 diabetes among young adults

Lammi, N., Moltchanova, E., Blomstedt, P., Eriksson, J. G., Taskinen, O., Sarti, C., Tuomilehto, J. and Karvonen, M.

Diabetologia. 2007; 50(12): 2433-8. IF 5.247

AIMS/HYPOTHESIS: The aim of this study was to examine the effects of birth order and parental age on the risk of type 1 and type 2 diabetes among Finnish individuals aged 15-39 years.

METHODS: Data on all cases of type 1 diabetes (n = 1,345) and type 2 diabetes (n = 1,072), diagnosed between 1992 and 1996, were collected from four sources: standardised national reports from diabetes nurses, the National Hospital Discharge Register, the Drug Prescription Register and the Drug Reimbursement Register. Information on matched controls and the family members of all study subjects were obtained from the National Population Registry. The odds ratios (ORs) for both types of diabetes were estimated using a conditional logistic regression model.

RESULTS: There was a U-shaped relationship between maternal age and the risk of type 2 diabetes in the offspring: the risk was higher in children born to young and old mothers compared with children born to mothers aged around 30 years. The children born second (OR 0.76, 95% CI 0.62-0.94), third (OR 0.73, 95% CI 0.55-0.95), or fourth (OR 0.66, 95% CI 0.47-0.94) had a lower risk of type 2 diabetes than the first-born children. Maternal age, paternal age, and birth order did not have an effect on the risk of type 1 diabetes in the individuals aged 15-39 years at the time of diagnosis.

CONCLUSIONS/INTERPRETATION: Maternal age and birth order are both associated with the risk of early-onset type 2 diabetes. However, part of these associations may be due to low birthweight. In this study neither parental age nor birth order showed a significant association with the risk of type 1 diabetes diagnosed after 15 years of age.

Coffee and Tea Consumption and Risk of Stroke Subtypes in Male Smokers

Larsson, S. C., Mannisto, S., Virtanen, M. J., Kontto, J., Albanes, D. and Virtamo, J.

Stroke. 2008. IF 5.391

BACKGROUND AND PURPOSE: Coffee and tea consumption could potentially reduce the risk of stroke because these beverages have antioxidant properties, and coffee may improve insulin sensitivity. We examined the associations of coffee and tea consumption with risk of stroke subtypes.

METHODS: We used prospective data from the Alpha-Tocopherol, Beta-Carotene Cancer Prevention Study, a cohort study of 26 556 male Finnish smokers aged 50 to 69 years without a history of stroke at baseline. Coffee and tea consumption was assessed at baseline using a validated food-frequency questionnaire. During a mean follow-up of 13.6 years, from 1985 through December 2004, 2702 cerebral infarctions, 383 intracerebral hemorrhages, and 196 subarachnoid hemorrhages were ascertained from national registries.

RESULTS: After adjustment for age and cardiovascular risk factors, both consumption of coffee and tea was statistically significantly inversely associated with the risk of cerebral infarction but not intracerebral or subarachnoid hemorrhage. The multivariate relative risk of cerebral infarction for men in the highest category of coffee consumption (>/=8 cups/d) was 0.77 (95% CI, 0.66 to 0.90; P for trend <0.001) compared with those in the lowest category (<2 cups/d). The corresponding relative risk comparing men in the highest category of tea consumption (>/=2 cups/d) with those in the lowest category (nondrinkers) was 0.79 (95% CI, 0.68 to 0.92; P for trend=0.002).

CONCLUSIONS: These results suggest that high consumption of coffee and tea may reduce the risk of cerebral infarction among men, independent of known cardiovascular risk factors.

Analytical techniques for drug detection in oral fluid

Lillsunde, P.

Ther Drug Monit. 2008; 30(2): 181-7.

ABSTRACT:: Analytical techniques for detection of drugs in oral fluid (OF) are reviewed with emphasis on applications used in European Union (EU) roadside testing projects. Oral fluid is readily accessible and collectible. It has become an interesting material because no medical personnel are needed for sampling. This matrix is especially applicable for preliminary drug testing in driving under the influence controls and for monitoring illicit drug use in drug treatment. Oral fluid is also an increasingly used specimen in epidemiologic studies and in workplace drug testing. Drugs are present at lower levels in OF than in urine. The window of detection of drugs in OF reflects the corresponding window in blood, suggesting OF as a specimen of choice for roadside testing. Saliva/blood ratios vary from drug to drug, from person to person, and even intraindividually making therapeutic drug monitoring in OF challenging. Several sensitive methods for drug testing in OF have been developed during the last years.

Probiotics in infancy induce protective immune profiles that are characteristic for chronic low-grade inflammation

Marschan, E., Kuitunen, M., Kukkonen, K., Poussa, T., Sarnesto, A., Haahtela, T., Korpela, R., Savilahti, E. and Vaarala, O.

Clinical and Experimental Allergy. 2008; 38(4): 611-618. Article. IF 3.668

Background Probiotics are widely studied both in the treatment and prevention of allergic diseases, but their mode of action is poorly known. Objective Our aim was to examine the effect of probiotic bacteria on in vivo cytokine, antibody, and inflammatory responses in allergy-prone infants. Methods In a randomized double-blind study, probiotic bacteria or placebo were given for 1 month before delivery to mothers and for 6 months to infants with a family history of allergy. Plasma samples were analysed for C-reactive protein (CRP), total IgA and IgE, food-specific IgA, IgG, and IgE, IL-2, IL-4, IL-6, IL-10, TNF-alpha, and IFN-gamma. We analysed the associations of immunological and inflammatory parameters at age 6 months with probiotic treatment and allergic phenotype at 2 years. Results Infants receiving probiotic bacteria had higher plasma levels of CRP (P=0.008), total IgA (P=0.016), total IgE (P=0.047), and IL-10 (P=0.002) than infants in the placebo group. Increased plasma CRP level at age 6 months was associated with a decreased risk of eczema [odds ratio (OR) 0.41 [95% confidence interval (CI) 0.17-0.99], P=0.046], and with a decreased risk of allergic disease [OR 0.38 (95% CI 0.16-0.87), P=0.023] at age 2 years, when adjusted with probiotic use. Conclusion The association of CRP with a decreased risk of eczema at 2 years of age in allergy-prone children supports the view that chronic, low-grade inflammation protects from eczema. Probiotic-induced low-grade inflammation was characterized by elevation of IgE, IgA, and IL-10, the changes typically observed in helminth infection-associated induction of regulatory mechanisms. The findings emphasize the role of chronic microbial exposure as an immune modulator protecting from allergy.

Social participation, trust and self-rated health: a study among ageing people in urban, semi-urban and rural settings

Nummela, O., Sulander, T., Rahkonen, O., Karisto, A. and Uutela, A.

Health Place. 2008; 14(2): 243-53. IF 1.828

This study examined associations between self-rated health and combinations of social participation and trust among ageing people in three living areas of Finland (N=2815, 66% response rate). Social participation and trust combinations were: low social capital (low participation/low trust), traditionalism (low/high), "the miniaturisation of community" (high/low) and high social capital (high/high). The highest rate of good self-rated health was found among the high social capital group, but after adjusting for background variables, statistical significance remained only in the urban area. High social capital measured at an individual level may thus promote health among ageing people.

Depression and its association with diabetes, cardiovascular disease, and birth weight

Paile-Hyvarinen, M., Raikkonen, K., Forsen, T., Kajantie, E., Yliharsila, H., Salonen, M. K., Osmond, C. and Eriksson, J. G.

Ann Med. 2007; 39(8): 634-40. IF 4.594

BACKGROUND: Diabetes increases the risk for depression.

AIM: To study the independent effects of diabetes mellitus (DM) and cardiovascular disease (CVD) on the prevalence of depression and to examine low birth weight as a possible common explanatory factor.

METHODS: 2003 subjects from the Helsinki Birth Cohort Study underwent a 75-g oral glucose tolerance test and filled out the Beck Depression Inventory.

RESULTS: Depressive symptoms were more prevalent among subjects with diabetes (23.5%) than among those with normal glucose tolerance (16.6%) (P < 0.001). A history of CVD also markedly increased the odds of having depressive symptoms (odds ratio (OR) = 2.38, 95% confidence interval (CI) = 1.70-3.32, P < 0.001). The association between DM and depressive symptoms was, however, rendered non-significant when adjusting for the presence of CVD. Being born with a low birth weight doubled the risk for having depressive symptoms (OR = 2.64, 95% CI = 1.42-4.91, P = 0.002) and magnified the association between CVD/DM and depression.

CONCLUSION: Diabetes has only a minor independent effect on concurrent occurrence of depressive symptoms, while cardiovascular disease seems to be a more important underlying factor. The association between disease and depression is in particular characteristic to individuals born with a low birth weight.

Roadside oral fluid testing: Comparison of the results of Drugwipe 5 and Drugwipe Benzodiazepines on-site tests with laboratory confirmation results of oral fluid and whole blood

Pehrsson, A., Gunnar, T., Engblom, C., Seppa, H., Jama, A. and Lillsunde, P.

Forensic Science International. 2008; 175(2-3): 140-148. Article. IF 1.397

Drugged drivers pose a serious threat to other people in traffic as well as to themselves. Reliable oral fluid screening devices for on-site screening of drugged drivers would be both a useful and convenient means for traffic control.

In this study we evaluated the appropriateness of Drugwipe 5 and Drugwipe Benzodiazepines oral fluid on-site tests for roadside drug screening. Drivers suspected of driving under the influence of drugs were screened with the Drugwipe tests. Oral fluid and whole blood samples were collected from the drivers and tested for amphetamine-type stimulant drugs, cannabis, opiates, cocaine and benzodiazepines by immunological methods, GC and GC-MS. The performance evaluations of the tests were made by comparing the results of the Drugwipe tests with laboratory GC-MS confirmation results of oral fluid or whole blood. In addition to the performance evaluations of the Drugwipe tests based on laboratory results, a questionnaire on the practical aspects of the tests was written for the police officers who performed the tests. The aim of the questionnaire was to obtain user comments on the practicality of the tests as well as the advantages and weak points of the tests. The results of the performance evaluations were: for oral fluid (sensitivity; specificity; accuracy) amphetamines (95.5%; 92.9%; 95.3%), cannabis (52.2%; 91.2%: 85.1%), cocaine (50.0%; 99.3%; 98.6%), opiates (100%; 95.8%; 95.9%), benzodiazepines (74.4%; 84.2%; 79.2%) and for whole blood accordingly, amphetamines (97.7%; 86.7%; 95.9%), cannabis (68.3%; 87.9%; 84.9%), cocaine (50.0%; 98.5%; 97.7%), opiates (87.5%; 96.9%; 96.6%) and benzodiazepines (66.7%; 87.0%; 74.4%). Although the Drugwipe 5 successfully detected amphetamine-type stimulant drugs and the police officers were quite pleased with the current features of the Drugwipe tests, improvements must still be made regarding the detection of cannabis and benzodiazepines. (C) 2007 Elsevier Ireland Ltd. All rights reserved.

Infant Growth and Hostility in Adult Life

Raikkonen, K., Pesonen, A. K., Heinonen, K., Lahti, J., Kajantie, E., Forsen, T., Osmond, C., Barker, D. J. and Eriksson, J. G.

Psychosom Med. 2008. IF 3.857

Objective: Hostility may confer a risk of cardiovascular disease and all-cause mortality, but why is uncertain. A common origin in suboptimal fetal and early postnatal life may lie beneath. This study tested whether prenatal and postnatal growth predicts hostility in adult life.

Methods: Women (n = 939) and men (n = 740) born in Helsinki, Finland, from 1934 through 1944 filled out the Cook-Medley Hostility Scale at an average age of 63.4 years. Growth was estimated from birth, child welfare clinic, and school records. Adult body size was measured in a clinic.

Results: Men and women who had higher levels of hostility in adulthood were born lighter and thinner, showed slower weight gain from birth to 6 months of age, were lighter throughout childhood (standardized regression coefficients (beta) <-0.05; 95% confidence intervals (95% CI), -0.14 to -0.00; p values <.05), but were heavier in adulthood (beta values > 0.06; 95% CIs, 0.02-0.14; p values <.01). They were also shorter from 6 months until the age of 1 year (beta values <-0.09; 95% CIs, -0.14 to -0.03; p values <.003), and tended to be shorter in adulthood (beta = -0.05; 95% CI, -0.09 to 0.00; p = .06). The latter effects were largely attributable to slower growth in stature from birth to 6 months (beta = -0.08; 95% CI, -0.14 to -0.02; p = .005). The associations were not explained by major confounders.

Conclusions: Our study suggests that slow prenatal and infant growth is linked with hostility in adult life.

What is ideal genetic counselling? A survey of current international guidelines

Rantanen, E., Hietala, M., Kristoffersson, U., Nippert, I., Schmidtke, J., Sequeiros, J. and Kääriäinen, H.

European Journal of Human Genetics. 2008; 16(4): 445-452. Article. IF 3.697

The objective of this article is to review guidelines that address counselling in the context of genetic testing in order to summarise what aspects of counselling they consider most important, and to examine how they construct the ideal of genetic counselling.

Guidelines were collected by examining the websites of different international professional, political, ethical and patient organisations, either previously known or found with the help of the Google search engine, and also using references listed in other studies. The most frequently mentioned topics in the collected 56 guidelines were sought, and this was carried out with the software package Qualitative Solutions and Research for Non-numerical Unstructured Data Indexing Searching and Theorizing. Topics related to genetic counselling that were mentioned in at least 30 of 56 collected documents were considered to be the most important aspects of genetic counselling. The ideal of genetic counselling is expressed in the analysed guidelines as being composed of (1) an appropriately trained professional who understands genetics and its ethical implications well; (2) relevant and objective information; (3) assurance of the counsellee's understanding; (4) psychological support; (5) informed consent; (6) confidentiality of genetic information; (7) considering familial implications; (8) appropriate handling of potential discrimination of testing; and (9) assuring autonomous decision-making by the counsellee.

The ideal of genetic counselling is rather consistent in the guidelines, but there are some contradictions between the requirements of objective information-giving and adapting counselling to counsellee's circumstances.

No association between common variants in glyoxalase 1 and autism spectrum disorders

Rehnstrom, K., Ylisaukko-Oja, T., Vanhala, R., von Wendt, L., Peltonen, L. and Hovatta, I.

Am J Med Genet B Neuropsychiatr Genet. 2008; 147(1): 124-7. IF 4.463

The autism spectrum disorders (ASDs) are complex diseases with a strong genetic component.

Numerous candidate gene studies have tested association between various functional and positional candidate genes and autism, but no common variation predisposing for autism has been identified to date. It has been previously proposed, that glyoxalase 1 (GLO1) might be involved in the pathogenesis of autism as GLO1 protein polarity was significantly changed in the brains of autism patients compared to controls. GLO1 harbors a functional polymorphism that affects the polarity and the enzymatic activity of the protein. In the same study, this polymorphism showed a suggestive association to autism. To investigate whether common variants in GLO1 predispose to autism in the Finnish population, we have genotyped six polymorphisms in GLO1 in families with more than 230 individuals affected with ASDs and carried out both linkage and association analyses. We did not observe significant linkage or association between any SNP and ASDs.

Therefore, we suggest that common variants in GLO1 are not significant susceptibility factors for ASDs in the Finnish population.

Ethically problematic treatment decisions in different medical specialties

Saarni, S. I., Halila, R., Palmu, P. and Vanska, J.

J Med Ethics. 2008; 34(4): 262-7. IF 1.222

BACKGROUND: Ethical dilemmas are an integral part of medicine. Whether physicians actually feel that they have made ethically problematic treatment decisions or choices in their work is largely unknown. Identifying physicians with ethical problems, and the types of problems and underlying factors, might benefit organisational and educational efforts to help physicians solve ethical dilemmas in a constructive way. We investigated how the frequency and types of ethically difficult treatment decisions vary by specialty.

METHOD: A mail survey of all non-retired Finnish physicians (n = 17,172, response rate 75.6%) was conducted in 2004. Of those who had made any ethically problematic treatment decisions, the types of decisions and reasons given for these decisions were asked for. Factor analysis was used to investigate clustering of ethically problematic treatment decisions, and logistic regression to investigate the effect of specialty, adjusted for age and gender.

RESULTS: Psychiatrists experienced ethically problematic treatment decisions most frequently, followed by pulmonologists, internists and neurologists. Problems were reported least often by pathologists, laboratory physicians and ophthalmologists. Overtreatment was more common than undertreatment in most specialties, with the exception of psychiatrists who emphasised undertreatment and patient rights issues.

CONCLUSION: Physicians of different specialties differ significantly regarding frequency and types of ethically problematic treatment decisions they have made. Psychiatrists differ from all other specialists in reporting more undertreatment and patient rights issues. Experiencing ethically problematic decisions might affect the quality of care and physician well-being in many ways. The findings could be useful for both under- and postgraduate ethics education.

Ethically problematic treatment decisions: a physician survey

Saarni, S. I., Parmanne, P. and Halila, R.

Bioethics. 2008; 22(2): 121-9. IF 1.672

BACKGROUND: Experiencing ethical problems requires both ethically problematic situations and ethical sensitivity. Ethically problematic treatment decisions are distressing and might reflect health care quality problems. Whether all physicians actually experience ethical problems, what these problems are and how they vary according to physician age, gender and work sector are largely unknown.

METHODS: A mail survey of all non-retired physicians licensed in Finland (n = 17,172, response rate 75.6%).

RESULTS: The proportion of physicians reporting having made ethically problematic treatment decisions decreased in linear fashion from 60% at ages below 30 years to 21% at ages over 63 years. The only problem that did not decrease in frequency with age was having withdrawn necessary treatments. Women and primary care physicians reported problematic decisions most often, although gender differences were small. Primary care physicians most often reported having performed too many investigations or having pressured patients, whereas hospital physicians emphasized having withdrawn necessary treatments. Performing unnecessary treatments or investigations was explained by pressure from patients or relatives, and performing too few treatments or investigations was explained by inadequate resources.

CONCLUSIONS: In general, young physicians felt pressured to do too much, whereas older physicians felt they could not do enough due to inadequate resources. Older physicians might be less exposed to ethically problematic situations, be more able to handle them or have lower ethical sensitivity. Young physicians could benefit from support in resisting pressure to perform unnecessary treatments, whereas older physicians might benefit from training in recognizing ethical issues.

Plasma ascorbic acid preparation and storage for epidemiological studies using TCA precipitation

Salminen, I. and Alfthan, G.

Clin Biochem. 2007. IF 2.331

OBJECTIVES:: To introduce a procedure to validate an ascorbic acid method using trichloroacetic acid (TCA) for plasma stabilization at different storage temperatures.

METHODS:: EDTA and heparin plasma were precipitated with TCA (1:5) containing 0.54 mol/L EDTA, or without. Samples were stored at -20 degrees C and -70 degrees C and their stability was tested at room temperature for 24 h.

RESULTS:: A significant 40% loss (p<0.001) of plasma ascorbic acid was found when EDTA samples with added EDTA were stored at -20 degrees C for 2-4 weeks compared with storage at -70 degrees C. Ascorbic acid in heparin plasma without added EDTA was most unstable and samples left at room temperature for 24 h lead to almost a total loss of ascorbic acid. Addition of EDTA to the TCA solution improved stability of samples of both plasma types at room temperature.

CONCLUSION:: The recommended procedure for ascorbic acid determination in plasma stabilized with TCA is immediate storage at -70 degrees C and inclusion of EDTA into the TCA solution.

Whole genome amplification with phi29 DNA polymerase to enable genetic or genomic analysis of samples of low DNA yield

Silander, K. and Saarela, J.

Methods Mol Biol. 2008; 439: 1-18.

In many large genetic studies, the amount of available DNA can be one of the criteria for selecting samples for study. In the case of large population cohorts, selecting samples based on their DNA yield can lead to biased sample selection. In addition, many valuable clinical and research sample collections exist in which the amount of DNA is very small. Unbiased whole genome amplification (WGA) of such unique samples enables genomewide scale genetic studies that would have been impossible otherwise. Multiply primed rolling circle amplification (MPRCA) and multiple displacement amplification (MDA) methods are based on the same principle. The DNA amplification is non-PCR based and uses Phi29 DNA polymerase and random hexamer primers for unbiased whole genome amplification. MDA is used for linear DNA molecules, such as genomic DNA.

This chapter reviews the various applications in which whole genome amplified DNA can be used, the types of commercial kits available, and the quality control steps necessary before using the DNA in the genetic studies.

Hepatitis C virus proteins interfere with the activation of chemokine gene promoters and downregulate chemokine gene expression

Sillanpaa, M., Kaukinen, P., Melen, K. and Julkunen, I.

J Gen Virol. 2008; 89(Pt 2): 432-43. IF 3.110

The hepatitis C virus (HCV) non-structural (NS) 3/4A protein complex inhibits the retinoic acid inducible gene I (RIG-I) pathway by proteolytically cleaving mitochondria-associated CARD-containing adaptor protein Cardif, and this leads to reduced production of beta interferon (IFN-beta).

This study examined the expression of CCL5 (regulated upon activation, normal T-cell expressed and secreted, or RANTES), CXCL8 (interleukin 8) and CXCL10 (IFN-gamma-activated protein 10, or IP-10) chemokine genes in osteosarcoma cell lines that inducibly expressed NS3/4A, NS4B, core-E1-E2-p7 and the entire HCV polyprotein. Sendai virus (SeV)-induced production of IFN-beta, CCL5, CXCL8 and CXCL10 was downregulated by the NS3/4A protein complex and by the full-length HCV polyprotein. Expression of NS3/4A and the HCV polyprotein reduced the binding of interferon regulatory factors (IRFs) 1 and 3 and, to a lesser extent, nuclear factor (NF)-kappaB (p65/p50) to their respective binding elements on the CXCL10 promoter during SeV infection. Furthermore, binding of IRF1 and IRF3 to the interferon-stimulated response element-like element, and of c-Jun and phosphorylated c-Jun to the activator protein 1 element of the CXCL8 promoter, was reduced when NS3/4A and the HCV polyprotein were expressed. In cell lines expressing NS3/4A and the HCV polyprotein, the subcellular localization of mitochondria was changed, and this was kinetically associated with the partial degradation of endogenous Cardif.

These results indicate that NS3/4A alone or as part of the HCV polyprotein disturbs the expression of IRF1- and IRF3-regulated genes, as well as affecting mitogen-activated protein kinase kinase- and NF-kappaB-regulated genes.

In vitro toxicity of cereulide on porcine pancreatic Langerhans islets

Virtanen, S. M., Roivainen, M., Andersson, M. A., Ylipaasto, P., Hoornstra, D., Mikkola, R. and Salkinoja-Salonen, M. S.

Toxicon. 2008. IF 2.509

Cereulide is a K(+) ionophore cytotoxic and mitochondriotoxic to primary cells and cell lines of human and other mammalian origins. It is a heat-stable, highly lipophilic (logK(ow) 5.96) peptide (1152gmol(-1)) produced by certain strains of Bacillus cereus, a bacterium connected to emetic food poisonings.

In this study the pancreatic toxicity of purified cereulide, and cereulide-containing bacterial extracts, was studied using fetal porcine Langerhans islets in culture. Exposure to 1ngml(-1) of purified cereulide caused necrotic cell death of the islet cells impairing their insulin content within 2 days. Cell extracts of cereulide-positive B. cereus strains connected to food poisoning or isolated from foodstuffs were toxic, corresponding to their measured cereulide content. Extracts of B. cereus strains producing or not producing the B. cereus diarrheal toxin, but no cereulide, were tolerated by the porcine islet cultures up to concentrations 1000-fold higher compared to extracts from strains containing cereulide, and up to exposure times of 7d. Cereulide thus was identified as the B. cereus-produced substance toxic towards porcine fetal Langerhans islets and beta cells.

Identification of Cytokines That Might Enhance the Promoter Activity of HLA-B27

Zhao, L., Fong, Y., Granfors, K., Gu, J. and Yu, D.

J Rheumatol. 2008. IF 2.940

OBJECTIVE: Although the mechanism by which HLA-B27 induces ankylosing spondylitis is unclear, a minimum threshold of transcription is essential, a process controlled at the promoter. Our aim was to scan the effect of a panel of cytokines on the promoter of the HLA-B27 gene over serial timepoints.

METHODS: The promoter region of B*2705 gene was cloned into a luciferase reporter, stably transfected into HeLa cells, and used to monitor the serial effect of 25 cytokines. Results of HLA-B27 promoter-reporter assays were compared to those of real-time polymerase chain reactions.

RESULTS: After an initial delay, significant activation of the HLA-B27 promoter was observed with tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma), and IFN-beta. While early response of the HLA-B27 promoter was highest with TNF-alpha and IFN-gamma, ultimately the highest activity was observed with IFN-beta.

CONCLUSION: The only promoter of HLA-B alleles studied in the past was that of HLA-B7, and always at only a single fixed timepoint of culture. This is the first study to show that activation of HLA-B27 promoter is a sequential event, and that TNF-alpha and IFN-beta are major participants at different timepoints.