Multilocus microsatellite typing as a new tool for discrimination of Leishmania infantum MON-1 strains

The Leishmania donovani complex, which consists of L. donovani, L. infantum-L. chagasi, and L. archibaldi, is responsible for visceral manifestations of leishmaniasis. Multilocus enzyme electrophoresis is the standard method for the characterization and identification of strains of Leishmania. For L. infantum, the predominance of zymodeme MON-1 significantly reduces the discriminative power of this approach. In the present study, we developed 17 independent polymorphic microsatellite markers for the typing of strains of L. infantum, with the main emphasis on zymodeme MON-1. The discriminative powers of 11 markers selected from among these markers were tested by using a panel of 63 isolates of the L. donovani complex. Unique multilocus genotypes were observed for the strains analyzed, with only three exceptions. Model-based and distance-based analyses of the data set showed comparable results. It was possible to discriminate between L. donovani sensu stricto, a non-MON-1 group of L. infantum isolates, and a MON-1 group of L. infantum isolates. Within MON-1, three clusters with geographical correlations became apparent. The frequency of heterozygosity in the alleles analyzed varied extremely between the different groups of isolates. The main clusters described are not consistent with species definitions based on isoenzyme analysis but confirm the results of former PCR-based investigations.     

Prediction of necessity for coronary artery revascularization by adenosine contrast-enhanced magnetic resonance imaging

BACKGROUND: An in-depth assessment of coronary heart disease (CHD) risk factors in Koreans was conducted to develop effective risk reduction strategies for this population. METHOD: Based on 2001 Korea National Health and Examination Survey, a cross-sectional survey, this paper presents the prevalence of major CHD risk factors and estimates future risk by applying the Framingham model and CHD risk factor guidelines of the National Cholesterol Education Program Adult Treatment Panel III (NCEP) to 4639 Koreans aged 20 to 79 years. RESULT: Our findings indicate that CHD is a major health threat to Koreans. Among these factors, low high-density lipoprotein cholesterol (HDL-C) was seen in 35.1% of men and 17.8% of women; both had a low rate of controlled high blood pressure (men 13.7%, women 23.6%), and age-adjusted current smoking was especially high (61.6%) in men. The 10-year CHD risk averaged 11.1% in men and 5.5% in women based on Framingham model, and 7.4% and 1.4% based on NCEP guidelines. About 9.1% of men and 2.6% of women were "high-risk," (>20%) based on the Framingham model, and 4.7% and 0.7%, respectively, on the NCEP guideline. Diabetes was the most prevalent risk factor among high-risk individuals and total cholesterol in men and low HDL-C in women was fairly high regardless of prediction method. CONCLUSION: These findings suggest that current coronary heart health of Koreans is nearly comparable to that of western developed countries. Developing and implementing effective population-based intervention strategies focusing on diabetes is warranted to lower the CHD risk for Koreans.     

Neuropsychology of bipolar disorders

In this article the contribution of neuropsychological research for a better understanding of the psychopathology of mood disorders is reviewed. First, the broad spectrum of bipolar disorders is described. Second, a selective review of important results of neuropsychological studies with patients with mood disorders is presented. Although several methodological problems limit the interpretation of the findings, there is evidence that patients with a bipolar disorder show a consistent impairment in attention, memory/learning and executive functions. The cognitive deficits are still visible during clinical recovery (euthymia) and closely associated with psychosocial limitation in daily life. Finally, the impact of neuropsychological findings is considered in relation to assessment, treatment and prognosis.     

Children in family mediation: A practice model

The authors briefly describe the history of family mediation under the perspective of the role of the children in the process of mediation. They state that originally children were not directly included. But through empirical studies and different higher escalated families asking for help by mediation, the inclusion of children got an important issue in theory and practice. The discussion began with the question in which phases of the mediation process the children should be included - it went to the issue of the age of the children - and the authors propose to take the amount of escalation in the family as the most important point of reference to decide if and how the children should be included. They suggest to diagnose the loss of responsibility and autonomy of the parents on a nine level scale (from F. Glasl) with the parents and to decide and negotiate with them how the children will be included. They describe five different settings of inclusion of the children.     

The human prion protein residue 129 polymorphism lies within a cluster of epitopes for T cell recognition

T cell immune responses to central nervous system-derived and other self-antigens are commonly described in both healthy and autoimmune individuals. However, in the case of the human prion protein (PrP), it has been argued that immunologic tolerance is uncommonly robust. Although development of an effective vaccine for prion disease requires breaking of tolerance to PrP, the extent of immune tolerance to PrP and the identity of immunodominant regions of the protein have not previously been determined in humans. We analyzed PrP T cell epitopes both by using a predictive algorithm and by measuring functional immune responses from healthy donors. Interestingly, clusters of epitopes were focused around the area of the polymorphic residue 129, previously identified as an indicator of susceptibility to prion disease, and in the C-terminal region. Moreover, responses were seen to PrP peptide 121-134 containing methionine at position 129, whereas PrP 121-134 [129V] was not immunogenic. The residue 129 polymorphism was also associated with distinct patterns of cytokine response: PrP 128-141 [129M] inducing IL-4 and IL-6 production, which was not seen in response to PrP 128-141 [129V]. Our data suggest that the immunogenic regions of human PrP lie between residue 107 and the C-terminus and that, like with many other central nervous system antigens, healthy individuals carry responses to PrP within the T cell repertoire and yet do not experience deleterious autoimmune reactions.     

Chronic caloric restriction partially protects against age-related alteration in serum metabolome

Calorie restriction (CR) remains the most robust metabolic intervention to extend lifespan and improve healthspan in several species. Using global and targeted mass spectrometry-based metabolomics approaches, here we show that chronic CR prevents age-related changes in specific metabolic signatures. Global metabolomic analysis using ultra-performance liquid chromatography–tandem mass spectrometry detected more than 7,000 metabolites in sera from ad-libitum-fed young, aged, and aged C57BL/6 mice maintained on 40 % CR. Multivariate statistical analysis of mass spectrometry data revealed a clear separation among the young, aged, and aged–CR mice demonstrating the potential of this approach for producing reliable metabolic profiles that discriminate based on age and diet. We have identified 168 discriminating features with high statistical significance (p ≤ 0.001) and validated and quantified three of these metabolites using targeted metabolite analysis. Calorie restriction prevented the age-related alteration in specific metabolites, namely lysophosphatidylcholines (16:1 and 18:4), sphingomyelin (d18:1/12:0), tetracosahexaenoic acid, and 7α-dihydroxy-4-cholesten-3-one, in the serum. Pathway analysis revealed that CR impacted the age-related changes in metabolic byproducts of lipid metabolism, fatty acid metabolism, and bile acid biosynthesis. Our data suggest that metabolomics approach has the potential to elucidate the metabolic mechanism of CR’s potential anti-aging effects in larger-scale investigations.

Electronic supplementary material

The online version of this article (doi:10.1007/s11357-012-9430-x) contains supplementary material, which is available to authorized users.     

Erythrocyte membrane cholesterol and lipid core growth in a rabbit model of atherosclerosis: Modulatory effects of rosuvastatin

Background

Lipid core expansion is partly responsible for the conversion of a stable atherosclerotic lesion to a rupture-prone plaque. Intraplaque hemorrhage contributes to the accumulation of cholesterol within unstable plaques. In the present study, we investigated, using a rabbit model of atherosclerosis, the extent to which diet-induced increases in cholesterol content of erythrocyte membranes (CEM) contribute to lipid core expansion and the modulatory effect of rosuvastatin use.

Methods and results

Rabbits fed with atherogenic diet (0.75% cholesterol) for 5 months exhibited advanced atherosclerotic lesions (mean plaque area, 0.39 ± 0.03 mm²), and lipid core size was associated with the concentration–time integral (CTI) of CEM levels (r = 0.567, P = 0.004) independent of other established predictors of lipid core size. Further experiments were performed by feeding rabbits atherogenic diet (1% cholesterol) for 3 months, followed by either normal diet or normal diet plus rosuvastatin for the next 3 months. Although no differences were observed in total plaque area between both groups, administration of rosuvastatin was associated with significantly smaller lipid cores, fewer macrophages within the lipid core, less microvessels as well as with lower CTI of CEM levels compared to normal diet alone. Moreover, intraplaque erythrocyte membranes covered a smaller lipid core area in rabbits under rosuvastatin plus normal diet as opposed to rabbits under diet alone.

Conclusions

Increased CEM levels, induced by high-cholesterol diet, are associated with lipid core growth. Ingestion of a potent HMG-CoA reductase inhibitor (rosuvastatin) may decrease CEM levels, and this effect may contribute to regression of the lipid core.     

Tensile bond strength of different adhesive systems between bone and composite compared: an in vitro study.

INTRODUCTION: The aim was to compare tensile bond strength of three dentine adhesive systems (Excite, Clearfil New Bond, Etch & Prime 3.0) and two cyanoacrylate adhesives (Cyano Veneer, Histoacryl) to animal bone in vitro. MATERIAL AND METHODS: Specimens from five porcine mandibles (diameter 9 mm) with a total thickness of 4mm (+/-0.5mm) and a cortical layer of 1.5mm (+/-0.2mm) were prepared using trephine burs under constant water cooling and under standardized conditions. They were assigned to five experimental groups. Tensile bond strength of five different adhesive agents (Clearfil New Bond, Etch & Prime 3.0, Excite, Histoacryl and Cyano Veneer was measured 15 min after application and after light curing of a composite material (Tetric Ceram, colour A2) added thereupon using a universal testing machine. Qualitative control was performed by scanning electron microscopy, while examining loaded and unloaded specimens. RESULTS: The measured tensile bond strength was as follows: Clearfil New Bond 8.00 MPa (+/-1.36 MPa), Etch & Prime 3.0 4.05 MPa (+/-1.52 MPa), Excite 2.96 MPa (+/-1.34 MPa), Histoacryl 5.22 MPa (+/-2.00 MPa), Cyano Veneer 4.56 MPa (+/-0.76 MPa). Clearfil New Bond showed significantly higher bond strength than the other four adhesives. Scanning electron microscopy analysis of unloaded specimens showed mixed modes of fracture. As regards the loaded specimens, no tag formation as known for dentine was found. CONCLUSION: Within the limitations of an in vitro investigation it can be concluded that dentine adhesive systems might be useful for bone bonding. Tensile bond strength of the dentine adhesive systems tested on bone is comparable to that evaluated for dentine in earlier investigations.     

Subclinical CNS Inflammation as Response to a Myelin Antigen in Humanized Mice

Multiple sclerosis is a demyelinating autoimmune disease of the CNS. Its animal model experimental autoimmune encephalomyelitis is commonly induced by active immunization with myelin antigens. To investigate human immune responses against myelin antigens in vivo we established a new subclinical experimental autoimmune encephalomyelitis model in humanized mice. NOD/Scidγc^?/? animals were transferred with peripheral blood mononuclear cells from healthy human donors and immunized with myelin antigens in complete Freund’s adjuvant and antigen-pulsed autologous dendritic cells. Human T cells recovered from these animals reacted specifically to the soluble domain of myelin oligodendrocyte glycoprotein and secreted proinflammatory cytokines. Furthermore, immunized animals developed subclinical CNS inflammation with infiltrating CD4⁺ and CD8⁺ T cells and production of encephalitogenic cytokines. Thus, this model of myelin-induced CNS inflammation by human T cells may allow testing of new human-specific therapeuticals for multiple sclerosis.