Cardiomyocyte-specific Loss of Diacylglycerol Acyltransferase 1 (DGAT1) Reproduces the Abnormalities in Lipids Found in Severe Heart Failure

Diacylglycerol acyltransferase 1 (DGAT1) catalyzes the final step in triglyceride synthesis, the conversion of diacylglycerol (DAG) to triglyceride. Dgat1^−/− mice exhibit a number of beneficial metabolic effects including reduced obesity and improved insulin sensitivity and no known cardiac dysfunction. In contrast, failing human hearts have severely reduced DGAT1 expression associated with accumulation of DAGs and ceramides. To test whether DGAT1 loss alone affects heart function, we created cardiomyocyte-specific DGAT1 knock-out (hDgat1^−/−) mice. hDgat1^−/− mouse hearts had 95% increased DAG and 85% increased ceramides compared with floxed controls. 50% of these mice died by 9 months of age. The heart failure marker brain natriuretic peptide increased 5-fold in hDgat1^−/− hearts, and fractional shortening (FS) was reduced. This was associated with increased expression of peroxisome proliferator-activated receptor α and cluster of differentiation 36. We crossed hDgat1^−/− mice with previously described enterocyte-specific Dgat1 knock-out mice (hiDgat1^−/−). This corrected the early mortality, improved FS, and reduced cardiac ceramide and DAG content. Treatment of hDgat1^−/− mice with the glucagon-like peptide 1 receptor agonist exenatide also improved FS and reduced heart DAG and ceramide content. Increased fatty acid uptake into hDgat1^−/− hearts was normalized by exenatide. Reduced activation of protein kinase Cα (PKCα), which is increased by DAG and ceramides, paralleled the reductions in these lipids. Our mouse studies show that loss of DGAT1 reproduces the lipid abnormalities seen in severe human heart failure.     

Roseovarius azorensis sp. nov., isolated from seawater at Espalamaca,Azores

A Gram-negative, motile, non-spore forming, rod shaped aerobic bacterium, designated strain SSW084^T, was isolated from a surface seawater sample collected at Espalamaca (38°33′N; 28°39′W), Azores. Growth was found to occur from 15 to 40 °C (optimum 30 °C), at pH 7.0–9.0 (optimum pH 7.0) and with 25–100 % seawater or 0.5–7.0 % NaCl in the presence of Mg²⁺ and Ca²⁺; no growth was found with NaCl alone. Colonies on seawater nutrient agar were observed to be punctiform, white, convex, circular, smooth, and translucent. Strain SSW084^T did not grow on Zobell marine agar and tryptic soy agar even when seawater supplemented. The major respiratory quinone was found to be Q-10 and the G + C content was determined to be 61.9 mol%. Phylogenetic analysis based on the 16S rRNA gene sequence indicated that strain SSW084^T belongs to the genus Roseovarius and that its closest neighbours are Roseovarius tolerans EL-172^T, Roseovarius mucosus DFL-24^T and Roseovarius lutimaris 112^T with 95.7, 95.4 and 95.3 % sequence similarity respectively. The remaining species of Roseovarius showed <95 % similarity. The polar lipids of strain SSW084^T were determined to be phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine, an unidentified lipid and one unidentified aminolipid. The major fatty acids identified were identified as C_18:1 ω7c (52.5 %) and C_16:0 (13.8 %). On the basis of phenotypic, molecular and chemotaxonomic characteristics, strain SSW084^T is considered to represent a novel species of the genus Roseovarius, for which Roseovarius azorensis sp. nov is proposed. The type strain is SSW084^T (=KCTC 32421^T = MTCC 11812^T).     

Microglial AGE-Albumin Is Critical in Promoting Alcohol-Induced Neurodegeneration in Rats and Humans

Alcohol is a neurotoxic agent, since long-term heavy ingestion of alcohol can cause various neural diseases including fetal alcohol syndrome, cerebellar degeneracy and alcoholic dementia. However, the molecular mechanisms of alcohol-induced neurotoxicity are still poorly understood despite numerous studies. Thus, we hypothesized that activated microglial cells with elevated AGE-albumin levels play an important role in promoting alcohol-induced neurodegeneration. Our results revealed that microglial activation and neuronal damage were found in the hippocampus and entorhinal cortex following alcohol treatment in a rat model. Increased AGE-albumin synthesis and secretion were also observed in activated microglial cells after alcohol exposure. The expressed levels of receptor for AGE (RAGE)-positive neurons and RAGE-dependent neuronal death were markedly elevated by AGE-albumin through the mitogen activated protein kinase pathway. Treatment with soluble RAGE or AGE inhibitors significantly diminished neuronal damage in the animal model. Furthermore, the levels of activated microglial cells, AGE-albumin and neuronal loss were significantly elevated in human brains from alcoholic indivisuals compared to normal controls. Taken together, our data suggest that increased AGE-albumin from activated microglial cells induces neuronal death, and that efficient regulation of its synthesis and secretion is a therapeutic target for preventing alcohol-induced neurodegeneration.     

Spontaneous asj-2J Mutant Mouse as a Model for Generalized Arterial Calcification of Infancy: A Large Deletion/Insertion Mutation in the Enpp1 Gene

Generalized arterial calcification of infancy (GACI), an autosomal recessive disorder caused by mutations in the ENPP1 gene, manifests with extensive mineralization of the cardiovascular system. The affected individuals in most cases die within the first year of life, and there is currently no effective treatment for this disorder. In this study, we characterized a spontaneous mutant mouse, asj-2J, as a model for GACI. These mice were identified as part of a phenotypic deviant search in a large-scale production colony of BALB/cJ mice at The Jackson Laboratory. They demonstrated a characteristic gait due to stiffening of the joints, with phenotypic similarity to a previously characterized asj (“ages with stiffened joints”) mouse, caused by a missense mutation in the Enpp1 gene. Complementation testing indicated that asj-2J and asj were allelic. PCR-based mutation detection strategy revealed in asj-2J mice a large, 40,035 bp, deletion spanning from intron 1 to the 3′-untranslated region of the Enpp1 gene, coupled with a 74 bp insertion. This was accompanied with a significant reduction in the plasma PP_i concentration and reduced PP_i/P_i ratio. As a consequence, extensive aberrant mineralization affecting the arterial vasculature, a number of internal organs, and the dermal sheath of vibrissae, a progressive biomarker of the ectopic mineralization process, was demonstrated by a combination of micro computed tomography, histopathology with calcium-specific stains, and direct chemical assay of calcium. Comparison of the asj and asj-2J mice demonstrated that the latter ones, particularly when placed on an acceleration diet high in phosphate and low in magnesium, had more extensive mineralization. Thus, the asj-2J mouse serves as a novel model for GACI, a currently intractable disorder.     

Full-Length Genomic Analysis of Korean Porcine Sapelovirus Strains

Porcine sapelovirus (PSV), a species of the genus Sapelovirus within the family Picornaviridae, is associated with diarrhea, pneumonia, severe neurological disorders, and reproductive failure in pigs. However, the structural features of the complete PSV genome remain largely unknown. To analyze the structural features of PSV genomes, the full-length nucleotide sequences of three Korean PSV strains were determined and analyzed using bioinformatic techniques in comparison with other known PSV strains. The Korean PSV genomes ranged from 7,542 to 7,566 nucleotides excluding the 3′ poly(A) tail, and showed the typical picornavirus genome organization; 5′untranslated region (UTR)-L-VP4-VP2-VP3-VP1-2A-2B-2C-3A-3B-3C-3D-3′UTR. Three distinct cis-active RNA elements, the internal ribosome entry site (IRES) in the 5′UTR, a cis-replication element (CRE) in the 2C coding region and 3′UTR were identified and their structures were predicted. Interestingly, the structural features of the CRE and 3′UTR were different between PSV strains. The availability of these first complete genome sequences for PSV strains will facilitate future investigations of the molecular pathogenesis and evolutionary characteristics of PSV.     

Circular RNAs in and out of Cells: Therapeutic Usages of Circular RNAs

RNAs are versatile molecules that are primarily involved in gene regulation and can thus be widely used to advance the fields of therapeutics and diagnostics. In particular, circular RNAs which are highly stable, have emerged as strong candidates for use on next-generation therapeutic platforms. Endogenous circular RNAs control gene regulatory networks by interacting with other biomolecules or through translation into polypeptides. Circular RNAs exhibit cell-type specific expression patterns, which can be altered in tissues and body fluids depending on pathophysiological conditions. Circular RNAs that are aberrantly expressed in diseases can function as biomarkers or therapeutic targets. Moreover, exogenous circular RNAs synthesized in vitro can be introduced into cells as therapeutic molecules to modulate gene expression networks in vivo. Depending on the purpose, synthetic circular RNA sequences can either be identical to endogenous circular RNA sequences or artificially designed. In this review, we introduce the life cycle and known functions of intracellular circular RNAs. The current stage of endogenous circular RNAs as biomarkers and therapeutic targets is also described. Finally, approaches and considerations that are important for applying the available knowledge on endogenous circular RNAs to design exogenous circular RNAs for therapeutic purposes are presented.     

Estimation of developmental parameters for adult emergence of Gonatocerus morgani,a novel egg parasitoid of the glassy-winged sharpshooter,and development of a degree-day model

The effects of temperature on the development (egg–adult emergence) of Gonatocerus morgani Triapitsyn, a newly-described parasitoid of Homalodisca vitripennis (Germar), were determined at 14.8, 18.7, 23.5, 26.9, 28.7, 30.4, 32.8, and 33.8 °C in the laboratory. Survival rate (percent adult emergence from parasitized host eggs) varied significantly among the experimental temperatures, with the highest (59%) and lowest (0%) occurring at 30.4 and 33.8 °C, respectively. The survival rates (%) were fitted with a polynomial model to describe a temperature-dependent pattern. Developmental rates (1/d) across seven temperatures were fitted with the nonlinear Briere model, which estimated the lower threshold to be 8.06 °C, the optimal temperature to be 29.22 °C, and the upper threshold to be 33.49 °C. A linear model fitted to developmental rates at 14.8–28.7 °C indicated that 189.75 degree-days above the lower threshold of 9.71 °C were required to complete development. A simulation model of G. morgani adult emergence was constructed to predict daily counts over the entire range of constant temperatures by incorporating the survival rate model, the Briere model, and the Weibull model. In outdoor validation, a degree-day model for predicting adult emergence showed ?2 d differences between prediction and observation. Based on the observed temperature requirement, the insect could complete thirteen to sixteen generations per year in southern California, depending on weather and location.