Treatment with apolipoprotein A-1 mimetic peptide reduces lupus-like manifestations in a murine lupus model of accelerated atherosclerosis

Introduction

The purpose of this study was to evaluate the effects of L-4F, an apolipoprotein A-1 mimetic peptide, alone or with pravastatin, in apoE^-/-Fas^-/-C57BL/6 mice that spontaneously develop immunoglobulin G (IgG) autoantibodies, glomerulonephritis, osteopenia, and atherosclerotic lesions on a normal chow diet.

Methods

Female mice, starting at eight to nine weeks of age, were treated for 27 weeks with 1) pravastatin, 2) L-4F, 3) L-4F plus pravastatin, or 4) vehicle control, followed by disease phenotype assessment.

Results

In preliminary studies, dysfunctional, proinflammatory high-density lipoproteins (piHDL) were decreased six hours after a single L-4F, but not scrambled L-4F, injection in eight- to nine-week old mice. After 35 weeks, L-4F-treated mice, in the absence/presence of pravastatin, had significantly smaller lymph nodes and glomerular tufts (P_{L, LP} < 0.05), lower serum levels of IgG antibodies to double stranded DNA (dsDNA) (P_L < 0.05) and oxidized phospholipids (oxPLs) (P_{L, LP} < 0.005), and elevated total and vertebral bone mineral density (P_{L, LP} < 0.01) compared to vehicle controls. Although all treatment groups presented larger aortic root lesions compared to vehicle controls, enlarged atheromas in combination treatment mice had significantly less infiltrated CD68⁺ macrophages (P_LP < 0.01), significantly increased mean α-actin stained area (P_LP < 0.05), and significantly lower levels of circulating markers for atherosclerosis progression, CCL19 (P_{L, LP} < 0.0005) and VCAM-1 (P_L < 0.0002).

Conclusions

L-4F treatment, alone or with pravastatin, significantly reduced IgG anti-dsDNA and IgG anti-oxPLs, proteinuria, glomerulonephritis, and osteopenia in a murine lupus model of accelerated atherosclerosis. Despite enlarged aortic lesions, increased smooth muscle content, decreased macrophage infiltration, and decreased pro-atherogenic chemokines in L-4F plus pravastatin treated mice suggest protective mechanisms not only on lupus-like disease, but also on potential plaque remodeling in a murine model of systemic lupus erythematosus (SLE) and accelerated atherosclerosis.     

Molecular architecture of the ribosome‐bound Hepatitis C Virus internal ribosomal entry site RNA

Internal ribosomal entry sites (IRESs) are structured cis‐acting RNAs that drive an alternative, cap‐independent translation initiation pathway. They are used by many viruses to hijack the translational machinery of the host cell. IRESs facilitate translation initiation by recruiting and actively manipulating the eukaryotic ribosome using only a subset of canonical initiation factor and IRES transacting factors. Here we present cryo‐EM reconstructions of the ribosome 80S‐ and 40S‐bound Hepatitis C Virus (HCV) IRES. The presence of four subpopulations for the 80S•HCV IRES complex reveals dynamic conformational modes of the complex. At a global resolution of 3.9 Å for the most stable complex, a derived atomic model reveals a complex fold of the IRES RNA and molecular details of its interaction with the ribosome. The comparison of obtained structures explains how a modular architecture facilitates mRNA loading and tRNA binding to the P‐site. This information provides the structural foundation for understanding the mechanism of HCV IRES RNA‐driven translation initiation.     

Weaning and the histology of the mandibular condyle in the rat.

Eighty-eight Long Evans/Turku rats were used in the study. The effect of the articulatory function on the mandibular condyle was observed histologically during normal growth, when the rat is changing its diet from milk to whole pellets as a part of weaning. Six animals each were killed at the age of 10, 15, 20, 25, 30, 35, 40 and 50 days for histological tissue processing. For further information, 30 animals were fed a soft diet (6 animals each were killed at the age of 25, 30, 35, 40 and 50 days), and 10 animals were fed hardened pellets (2 animals each were killed at the ages of 25, 30, 35, 40 and 50 days). An even and regular transition from mesenchymal cells via immature chondroblasts into mature chondroblasts and hypertrophied chondrocytes was found at 10, 15 and 20 days during normal growth and also at 25, 30, 35, 40 and 50 days when animals were fed a soft diet. This maturing process appeared to be disturbed at the age of 25, 30, 35 and 40 days in the superior aspect of the condyle in animals fed ordinary pellets. The density of the mesenchymal cell layer was decreased, and the amount of intercellular matrix seemed to be evaluated in mesenchymal and intermediate cell layers. These features were later manifest deeper in the cartilage as acellular regions and as cell clusters. The changes were similar but more severe when the animals were fed hardened pellets.(ABSTRACT TRUNCATED AT 250 WORDS)     

Hormone-regulated expression and distribution of versican in mouse uterine tissues

Background  

Remodeling of the extracellular matrix is one of the most striking features observed in the uterus during the estrous cycle and after hormone replacement. Versican (VER) is a hyaluronan-binding proteoglycan that undergoes RNA alternative splicing, generating four distinct isoforms. This study analyzed the synthesis and distribution of VER in mouse uterine tissues during the estrous cycle, in ovariectomized (OVX) animals and after 17beta-estradiol (E2) and medroxyprogesterone (MPA) treatments, either alone or in combination.     

The Associations of Objectively Measured Physical Activity and Sedentary Time with Cognitive Functions in School-Aged Children

Low levels of physical activity among children have raised concerns over the effects of a physically inactive lifestyle, not only on physical health but also on cognitive prerequisites of learning. This study examined how objectively measured and self-reported physical activity and sedentary behavior are associated with cognitive functions in school-aged children. The study population consisted of 224 children from five schools in the Jyväskylä school district in Finland (mean age 12.2 years; 56% girls), who participated in the study in the spring of 2011. Physical activity and sedentary time were measured objectively for seven consecutive days using the ActiGraph GT1M/GT3X accelerometer. Self-reported moderate to vigorous physical activity (MVPA) and screen time were evaluated with the questions used in the “WHO Health Behavior in School-aged Children” study. Cognitive functions including visual memory, executive functions and attention were evaluated with a computerized Cambridge Neuropsychological Test Automated Battery by using five different tests. Structural equation modeling was applied to examine how objectively measured and self-reported MVPA and sedentary behavior were associated with cognitive functions. High levels of objectively measured MVPA were associated with good performance in the reaction time test. High levels of objectively measured sedentary time were associated with good performance in the sustained attention test. Objectively measured MVPA and sedentary time were not associated with other measures of cognitive functions. High amount of self-reported computer/video game play was associated with weaker performance in working memory test, whereas high amount of computer use was associated with weaker performance in test measuring shifting and flexibility of attention. Self-reported physical activity and total screen time were not associated with any measures of cognitive functions. The results of the present study propose that physical activity may benefit attentional processes. However, excessive video game play and computer use may have unfavorable influence on cognitive functions.     

Mechanism of adaptation in the mandibular condyle of the mouse. An organ culture study

The cranial base and whole mandible of 7-day-old mice were cultured for 1 or 2 weeks in a novel organ culture system that provided an articulating movement for the temporomandibular joints. The mandibles were articulating either in a closed or in an open position to simulate the in vivo mouth-breathing pattern. The reactions of the condylar processes were followed macroscopically and microscopically after osteoid or von Kossa staining. After 1 week of culture, small differences were found in the shape of the condylar processes between the two groups. After 2 weeks, significantly increased growth was observed superiorly in those condyles which had been working in a closed position and posteriorly in the condyles which had been working in an open position. Histologic analysis revealed that after 1 week calcification had proceeded faster in the posterior aspect of the condyles in the mandibles which had been working in a closed position as compared to the mandibles which had been working in an open position, and the difference was even more marked after 2 weeks. In the latter condyles calcification had proceeded markedly in the anterosuperior aspect. Osteoid staining showed that perichondrial mesenchymal cells had differentiated into osteoblasts in the posterior aspect of the condyles in those mandibles which had been working in a closed position. This led to a ceasing of expansive growth in the posterior aspect and thus to a more upwardly directed condylar growth. In mandibles which had been working in the open position to simulate the mouth-breathing pattern in vivo, chondrogenesis and thus expansive growth continued in the posterior aspect, leading to a more posteriorly directed condylar growth.(ABSTRACT TRUNCATED AT 250 WORDS)     

<Emphasis Type="Italic">Chlamydomonas reinhardtii</Emphasis> chloroplasts express an orally immunogenic protein targeting the p210 epitope implicated in atherosclerosis immunotherapies

Key message

An algae-based vaccine model against atherosclerosis was developed with positive findings in terms of antigen yield and immunogenicity in mouse.

Abstract

Several immunotherapies against atherosclerosis have been evaluated at the preclinical level thus far, with some of them currently under evaluation in clinical trials. In particular, the p210 epitope from ApoB100 is known to elicit atheroprotective responses. Considering that Chlamydomonas reinhardtii is an attractive host for the production and delivery of subunit vaccines, in this study a chimeric protein consisting of the B subunit of the cholera toxin and the p210 epitope from ApoB100 (CTB:p210) has been expressed in C. reinhardtii chloroplast as an attempt to establish an oral vaccine candidate against atherosclerosis. The Chlamydomonas-made CTB:p210 protein was successfully expressed at levels of up to 60 µg per g of fresh weight biomass. The antigenic activity of the CTB and the p210 moiety was preserved in the CTB:p210 chimera. Moreover the algae-made CTB:p210 showed an immunogenic activity, when orally administered to BALB/c mice, as evidenced the presence of anti-p210 serum antibodies in mice treated with the algae-derived CTB:p210. The antibody response lasts for at least 80 days after the last boost. This experimental model is proposed as a convenient tool in the development of low cost atherosclerosis vaccines of easy compliance and friendly delivery. Further studies will determine the therapeutic potential of this algae-made vaccine in atherosclerosis animal models.