2.1.2006

New quinolone resistance phenomenon in Salmonella enterica: nalidixic acid-susceptible isolates with reduced fluoroquinolone susceptibility

Hakanen, A. J., Lindgren, M., Huovinen, P., Jalava, J., Siitonen, A. and Kotilainen, P.

J Clin Microbiol. 2005; 43(11): 5775-8.

We describe the emergence of a new quinolone resistance pattern in Salmonella enterica isolates from Southeast Asia. These isolates are susceptible to nalidixic acid but exhibit reduced susceptibility to ciprofloxacin. The increase of such strains may threaten the value of the nalidixic acid disk test to screen for reduced fluoroquinolone susceptibility in salmonellas.

Major depressive episode related to long unemployment and frequent alcohol intoxication

Hämäläinen, J., Poikolainen, K., Isometsä, E., Kaprio, J., Heikkinen, M., Lindeman, S. and Aro, H.

Nordic Journal of Psychiatry. 2005; 59(6): 486-491.

We studied the association between two major problems - unemployment and major depressive episode - and the impact of different timing of periods of unemployment and risk factors, especially alcohol intoxication, for major depressive episode among the unemployed. Major depressive episode during the last 12 months, plus current and past employment status and frequency of alcohol intoxication, were assessed within the nationally representative, cross-sectional 1996 Finnish Health Care Survey, in which non-institutionalized individuals aged 15 - 75 years were interviewed by using the Short Form of the University of Michigan version of the Composite International Diagnostic Interview (the UM-CIDI Short Form). Of the 5993 subjects interviewed, 3818 (64%) were occupationally active and included in the logistic regression analysis, showing that even after adjusting for other potentially confounding variables, current unemployment was associated with major depressive episode (odds ratio, OR = 1.78, 95% confidence interval, CI, 1.38 - 2.29). Further analysis revealed that the increased risk of major depressive episode was only related to long-term unemployment. Frequent alcohol intoxication (at least once a week) increased the risk of major depressive episode remarkably. Compared with the group ''Constantly employed, no frequent alcohol intoxication'', long-term unemployment with no frequent alcohol intoxication had moderately increased risk of major depressive episode (OR = 1.72 ( 95% CI 1.29 - 2.30) and those with frequent alcohol intoxication had highly increased risk [OR = 11.27 (95% CI 5.51 - 23.09) vs. OR - 1.72 (95% CI 1.29 - 2.30]. Long-term unemployment is associated with increased risk of major depressive episode. Frequent alcohol intoxication among long-term unemployed individuals greatly increases the risk of depression.

Selected ion monitoring in quantitative gas-liquid chromatographic - mass spectrometric detection of fatty acid methyl esters from environmental samples

Kontro M, Korhonen L, Vartiainen T, Pellikka P, Martikainen PJ.

J Chromatogr B Analyt Technol Biomed Life Sci. 2005.

To calculate selected ion monitoring (SIM) gas-liquid chromatography (GLC)-mass spectrometry (MS) results of phospholipid fatty acids (PLFAs) from environmental samples, coefficients were calculated for each fatty acid by dividing the sum of ion intensities in SCAN with that of ions followed in SIM. The SIM chromatogram areas were multiplied with the coefficients, and then processed as in SCAN. The results were compared to those obtained using calibration curves and SCAN. The calibration curve and coefficient based results had the greatest errors of 7.8 and 6.7%, respectively, outside standard deviations of SCAN percentages. The PLFA contents calculated using calibration curves and coefficients were 104.9+/-7.3% and 101.5+/-8.6%, respectively, of SCAN values. SIM increased sensitivity approximately 10-fold from SCAN, and the smallest detectable injected amount was approximately 50ng (0.18nmol) for 20 fatty acids, corresponding to 4x10(6) cells.

Neural basis of alcohol dependence

Kuoppasalmi, K. I.

Nordic Journal of Psychiatry. 2005; 59(5): 402-402. Meeting Abstract

Low circulating soluble interleukin 2 receptor level predicts rapid response in patients with refractory rheumatoid arthritis treated with infliximab

Kuuliala A, Nissinen R, Kautiainen H, Repo H, Leirisalo-Repo M

Annals of the Rheumatic Diseases. 2006; 65(1): 26-29.

Background: Treatment with infliximab induces a rapid therapeutic response in most patients with active rheumatoid arthritis. Factors predicting good response are not well known.

Objective: To study the predictive value of baseline level of soluble interleukin 2 receptor (sIL2R), a marker of lymphocyte activation, on the treatment response. Methods: 24 patients with active rheumatoid arthritis received intravenous infusions of infliximab at study entry, at two weeks, at six weeks, and at eight week intervals thereafter.

Outcome was evaluated at six weeks and 22 weeks. Clinical assessment and standard laboratory tests were made and the DAS28 disease activity score was calculated. Serum sIL2R level at entry was measured by automated immunoassay analyser (Immulite (R)). The mean change in DAS28 score from entry to six weeks and 22 weeks was calculated and related to sIL2R level using baseline adjusted robust regression analysis.

Results: Baseline level of serum sIL2R (mean (SD), 621 (325) U/ml) did not correlate with baseline DAS28 score (r = 0.24 (95% confidence interval, -0.18 to 0.58)). At six weeks DAS28 scores improved, with a mean change of -2.53 (-3.08 to -1.98) (p < 0.001). This change was predicted by low baseline sIL2R level (regression coefficient per 100 U/ml: 0.205 (0.003 to 0.407) (p = 0.047)). At 22 weeks the DAS28 scores improved, with a mean change of -2.26 (-2.75 to -1.77) (p < 0.001). The change was not predicted by baseline sIL2R level.

Conclusions: Low baseline sIL2R level may predict a rapid clinical response in patients with refractory rheumatoid arthritis treated with infliximab.

In vitro activities of 11 fluoroquinolones against 226 Campylobacter jejuni strains isolated from Finnish patients, with special reference to ciprofloxacin resistance

Lehtopolku, M., Hakanen, A. J., Siitonen, A., Huovinen, P. and Kotilainen, P.

Journal of Antimicrobial Chemotherapy. 2005; 56(6): 1134-1138.

Objectives: Although Campylobacter jejuni is naturally susceptible to fluoroquinolones, the resistance to these antimicrobials has increased rapidly during the recent years.

The aim of this study was to compare the activities of various older and newer fluoroquinolones towards C. jejuni, with special attention on ciprofloxacin-resistant strains. Methods: We analysed the in vitro activities of 11 fluoroquinolones against 226 C. jejuni strains collected from Finnish patients between 1995 and 2000.

Results: Of all 226 C. jejuni strains, 134 (59.3%) were resistant to ciprofloxacin (MIC >= 4 mg/L), 1 (0.4%) was intermediately resistant (MIC = 2 mg/L) and 91 (40.3%) were ciprofloxacin-susceptible (MIC <= 1 mg/L). The MIC50 and MIC90 values of ciprofloxacin for the 91 ciprofloxacin-susceptible strains were 0.25 and 0.5 mg/L, respectively. The corresponding MIC50 and MIC90 values of levofloxacin were 0.25 and 0.5 mg/L, and those of moxifloxacin were 0.125 and 0.25 mg/L, these being lower than those of norfloxacin and ofloxacin. The two newer fluoroquinolones, sitafloxacin and clinafloxacin, exhibited the lowest MIC50 and MIC90 values: 0.016 and 0.064 mg/L of sitafloxacin and 0.032 and 0.125 mg/L of clinafloxacin, respectively. Sitafloxacin and clinafloxacin exhibited the lowest MIC50 and MIC90 values also for the 134 ciprofloxacin-resistant C. jejuni strains: 0.25 and 1 mg/L of sitafloxacin and 1 and 4 mg/L of clinafloxacin, respectively.

Conclusions: Of the newer fluoroquinolones presently under development, sitafloxacin is in vitro highly effective towards C. jejuni, with low MIC values also for the ciprofloxacin-resistant strains. Sitafloxacin might be a candidate for clinical trials on campylobacteriosis.

Chronic systemic endotoxin exposure: an animal model in experimental hepatic encephalopathy

Lindros, K. O. and Järveläinen, H. A.

Metab Brain Dis. 2005; 20(4): 393-8.

Plasma levels of gut-derived endotoxins (lipopolysaccharides, LPS) are often elevated in cirrhotics and are thought to contribute to hepatic encephalopathy. Circulating LPS activates macrophages to produce tumor necrosis factor alpha (TNF-alpha) and other potentially cytotoxic proinflammatory mediators. A pathogenic role forendotoxins is supported by studies showing that treatment with Lactobacillusor antibiotics, both of which reduce LPS-producingintestinal Gram-negative bacteria, alleviates experimental liver damage. To mimic the "leaky gut" syndrome with endotoxin translocation into the circulation in cirrhotics, a new animal model was developed. Rats were chronically exposed to ethanol and for the four last weeks also infused with endotoxin into the jugular vein from subcutaneously implanted osmotic minipumps. Animals receiving endotoxin had elevated hepatic expression of both pro- and anti-inflammatory cytokines, but compared to ethanol treatment alone hepatic steatosis and inflammatory changes were only marginally increased. This demonstrates marked endotoxin tolerance, probably as a consequence of a counteracting anti-inflammatory cytokine response. The role of gut-derived endotoxin in hepatic encephalopathy has recently received considerable attention. To further delineate the role and actions of endotoxin and its extrahepatic effects, studies applying both acute challenge and chronic infusion seem warranted. The chronic endotoxin model, mimicking the "leaky gut," may best be combined with more robust ways to impair liver function, such as carbon tetrachloride treatment, bile duct ligation, or galactosamine administration.

In vitro and in vivo release of ciprofloxacin from osteoconductive bone defect filler

Mäkinen, T. J., Veiranto, M., Lankinen, P., Moritz, N., Jalava, J., Törmälä, P. and Aro, H. T.

Journal of Antimicrobial Chemotherapy. 2005; 56(6): 1063-1068.

Objectives: Impregnation of antimicrobial agents within biodegradable carriers with osteoconductive properties could provide the means for one-stage surgical treatment of osteomyelitis. In this study, the in vitro and in vivo antibiotic release from this type of bone defect filler was characterized.

Methods: Cylindrical pellets (2.5 x 1.5 mm) were manufactured from bioabsorbable poly(l-lactide-co-glycolide) (PLGA) matrix, ciprofloxacin [8.3 +/- 0.1% (w/w)] and osteoconductive bioactive glass microspheres (90-125 mu m) [27 +/- 2% (w/w)]. In vitro studies were carried out to delineate the release profile of the antibiotic. The antimicrobial activity of the release antibiotic was verified with MIC testing. In a time-sequence study in the rabbit, pellets were surgically implanted in the proximal tibia and the antibiotic concentrations achieved in bone were measured at 1, 2, 3, 4, 5 and 6 months.

Results: In vitro elution studies showed sustained release of ciprofloxacin at a therapeutic level (> 2 mu g/mL) over a time period of 4 months. The released ciprofloxacin had maintained its antimicrobial capacity against five standard ATCC strains. In vivo, the delivery system produced high local bone concentrations (247.9 +/- 91.0 mu g/g of bone) for a time period of 3 months with no significant systemic exposure. Histomorphometry and micro-CT imaging confirmed new bone formation around the pellets within 3 months as a sign of an independent osteoconductive property of the composite.

Conclusions: The tested composite seems to be a promising option for local therapy of surgically treated bone infections. The main advantages are the antibiotic release for a definite time period with therapeutic concentrations, which may minimize slow residual release at suboptimal concentrations.

Cow milk is not responsible for most gastrointestinal immune-like syndromes - evidence from a population-based study(1-3)

Paajanen, L., Korpela, R., Tuure, T., Honkanen, J., Järvelä, I., Ilonen, J., Knip, M., Vaarala, O. and Kokkonen, J.

American Journal of Clinical Nutrition. 2005; 82(6): 1327-1335.

Background: Gastrointestinal hypersensitivity to cow milk (CM) may be more common among school-aged children and young adults than previously thought.

Objective: The objective was to study various gastrointestinal complaints and the immunologic mechanisms associated with food-related. especially CM-related, gastrointestinal disorders in young adults. Design: Of 827 subjects aged 16-21 y who completed a questionnaire on food-related gastrointestinal symptoms, 49 symptomatic subjects agreed to a clinical examination, including an interview, blood tests, a lactose-maldigestion test, a blinded CM challenge and, in severely symptomatic subjects (n = 12), an endoscopic examination. Twenty-nine subjects served as controls.

Results: Approximately 10% of the subjects reported having major gastrointestinal symptoms, mainly food-related (n = 70 of 86), during the preceding year. Specific organic disease was found in 2 symptomatic subjects: 1 case of celiac disease and 1 of colitis. The result of the lactose-maldigestion test was positive in 16 of the remaining 47 symptomatic subjects, but only 4 carried the C/C-13910 genotype for adult-type hypolactasia. The symptomatic subjects had restricted their consumption of certain foods, particularly CM. However, in a blinded challenge, CM-induced symptoms were rare. The symptomatic subjects had higher plasma soluble intercellular adhesion molecule 1 (P = 0.007) and lower granzyme A (P = 0.001) concentrations than did the control subjects. Duodenal biopsy samples tended to have higher intraepithelial CD3+ cell counts (P 0.065) and a higher expression of transforming growth factor 0 (P 0.073) and interleukin 12p35 messenger RNA (P = 0.075) than did the control subjects.

Conclusions: In an unselected cohort of young adults, 8% reported food-related gastrointestinal symptoms. The finding of immunologic activity implied the existence of a food-related gastrointestinal syndrome but not one induced by CM.

A whole cell immunization-derived monoclonal antibody that protects cells from coxsackievirus A9 infection binds to both cell surface and virions

Roivainen, M., Alakulppi, N., Ylipaasto, P., Eskelinen, M., Paananen, A., Airaksinen, A. and Hovi, T.

J Virol Methods. 2005; 130(1-2): 108-16.

Coxsackievirus A9 (CAV-9) infects human rhabdomyosarcoma (RD) cells using an unidentified RGD-independent receptor. Monoclonal antibodies were prepared by immunizing mice with intact RD cells and by selecting cells from the cytopathic effect of CAV-9 for protection. Here we describe a monoclonal antibody that binds to host cell plasma membrane and protects cells from virus infection. In addition, binding of the virus to cell monolayers was more efficient in the presence of the antibody, suggesting that the antibody is also capable of recognizing virus particles. Immunoprecipitation and electron microscopy studies with highly purified virus preparations verified binding of the monoclonal antibody to the virus particles. The antibody also recognized coxsackievirus A21 and all three serotypes of poliovirus, but without affecting their infectivity. The amino acid sequence of CAV-9 recognized by the monoclonal antibody was identified by peptide mapping and by producing escape mutants in the presence of the antibody.

A heterogeneity-based genome search meta-analysis for autism-spectrum disorders

Trikalinos, T. A., Karvouni, A., Zintzaras, E., Ylisaukko-oja, T., Peltonen, L., Järvelä, I. and Ioannidis, J. P. A.

Molecular Psychiatry. 2006; 11(1): 29-36.

Autism and autism-spectrum disorders exhibit high heritability, although specific susceptibility genes still remain largely elusive. We performed a heterogeneity-based genome search meta-analysis (HEGESMA) of nine genome scans on autism or autism-spectrum disorders. Each genome scan was separated in 30 cM bins and the maximum linkage statistic from each bin was ranked. Significance for each bin's average rank and for between-scan heterogeneity (dis-similarity in the average ranks) was obtained through Monte Carlo tests. For autism, data from 771 affected sibpairs were synthesized across six separate genome scans. Region 7q22 - q32 reached genome-wide significance both in weighted and unweighted analyses, with evidence for significantly low between-scan heterogeneity. The flanking chromosomal region 7q32-qter reached the less stringent threshold of suggestive significance, with no evidence for low between-scan heterogeneity. For autism-spectrum disorders ( 634 affected sibpairs from five separate scans), no chromosomal region reached genome-wide significance. However, suggestive significance was reached for the chromosomal regions 17p11.2 - q12 and 10p12 - q11.1 in weighted analyses. There was evidence for significantly high between-scan heterogeneity for the former region. The meta-analysis suggests that the 7q22 - q32 region should be further scrutinized for autism susceptibility genes, while autism-spectrum disorders seem to have quite diverse linkage signals across scans, possibly suggesting genetic heterogeneity across subsyndromes and subpopulations.

Undiagnosed silent coeliac disease: A risk for underachievement?

Verkasalo, M. A., Raitakari, O. T., Viikari, J., Marniemi, J. and Savilahti, E.

Scandinavian Journal of Gastroenterology. 2005; 40(12): 1407-1412.

Objective. Silent coeliac disease is reported in 1% of Caucasian populations, but there is a lack of knowledge of its natural course and the risk of complications. The need for population screening is debated. We sought for complications of untreated coeliac disease in a well- defined cohort of Finnish adults.

Material and methods. Subjects ( n = 2427, ages 24 - 39 years) attending the 21- year follow- up visit of the study " Cardiovascular Risk in Young Finns'' completed an extensive questionnaire on their health, diet, social situation and family life, and were given a medical examination. Measurement of serum IgA- transglutaminase and IgA- endomysium antibodies identified 21 subjects with silent coeliac disease.

Results. The subjects with silent coeliac disease did not differ from the rest of the cohort in age, gender, stature, weight, medical diagnoses ( autoimmune, malignant), health concerns, use of alternative medications, physical activity, or in the cause of death their parents. They had lower serum HDL- cholesterol ( 1.12 versus 1.29 mmol/ L; p = 0.015), as described for active coeliac disease. Fewer ( 5.3% versus 22.8%; p = 0.047) had a university or college degree or worked in managerial or professional positions ( 28% versus 45%; p = 0.112).

Conclusions. The underachievement in education and working life observed in subjects with silent coeliac disease is a new and intriguing finding and may be related to the increased prevalence of depressive and disruptive behavioural disorders described in teenagers with untreated coeliac disease. Our findings add a new ingredient to the ongoing discussion regarding the need for population screening for silent coeliac disease.

Functional activity of antibodies after immunization of Finnish and Israeli infants with an 11-valent pneumococcal conjugate vaccine

Wuorimaa, T. K., Dagan, R., Bailleux, F., Haikala, R., Ekström, N., Eskola, J., Yaich, M. and Käyhty, H.

Vaccine. 2005; 23(46-47): 5328-32.

Finnish and Israeli infants received an 11-valent mixed carrier pneumococcal conjugate vaccine (11PCV) with or without aluminum adjuvant at the age of 2, 4, 6, and 12 months. We measured opsonophagocytic activity (OPA) of antibodies to pneumococcal strains of serotypes 4, 6B, 14, 19F, and 23F. At 7 months, OPA was clearly detected for all the serotypes. At 13 months, OPAs increased further and the proportion of individuals with a positive OPA ranged between 81 and 100%. The adjuvant improved functional activity of antibodies to serotype 6B pneumococci. In conclusion, immunization of infants with the 11PCV induced functionally active antibodies.