Aging is a significant risk factor for several diseases. Studies have uncovered multiple signaling pathways that modulate aging, including insulin/insulin-like growth factor-1 signaling (IIS). In Caenorhabditis elegans, the key regulator of IIS is DAF-16/FOXO. One of the kinases that affects DAF-16 function is the AMPK catalytic subunit homolog AAK-2. In this study, we report that PRY-1/Axin plays an essential role in AAK-2 and DAF-16-mediated regulation of life span. The pry-1 mutant transcriptome contains many genes associated with aging and muscle function. Consistent with this, pry-1 is strongly expressed in muscles, and muscle-specific overexpression of pry-1 extends life span, delays muscle aging, and improves mitochondrial morphology in AAK-2-DAF-16-dependent manner. Furthermore, PRY-1 is necessary for AAK-2 phosphorylation. Taken together, our data demonstrate that PRY-1 functions in muscles to promote the life span of animals. This study establishes Axin as a major regulator of muscle health and aging.

Purpose: Exosomes containing proteins, microRNAs and mRNAs are found in extracellular spaces such as blood and other body fluids and function as messengers in cell-cell communication through transfer of their molecular content. We hypothesize that exosomes containing miRNAs function in a novel communication mechanism among joint cells in osteoarthritis (OA) pathogenesis. We previously observed that the balance of anabolic and catabolic gene expression in chondrocytes was impaired by exosomes derived from OAsynovial fibroblasts (SFB). Angiogenesis in various joint tissues, including synovium, ligaments, menisci and the osteochondral junction, is a recently recognized important factor in OA pathogenesis. The purpose of this study was to investigate potential angiogenic function of exosomes from OA-SFB. Methods: Human SFB were obtained from a normal, OA and rheumatoid arthritis (RA) knee joints, and cultured with or without interleukin-1b (IL-1b). Exosomes were isolated by ExoQuick from IL-1b stimulated medium and control medium. Isolated exosomes fraction was measured for protein content using BCA protein assay kit. To examine the angiogenic functions of exosomes derived from SFB, we examined whether migration and tube formation in human umbilical vein endothelial cells (HUVECs) were affected by normal-, OA-, RA-SFB derived exosomes. Migration assay was evaluated using HUVECs in a modified Boyden chamber. In the tube formation assay, HUVECs were seeded into plates coated with growth-factor-reduced Matrigel and tube length was measured. miRNA profiles in the exosome preparations were established using 3D-Gene miRNA microarray. Results: Migration and tube formation activity were significantly higher in endothelial cells treated with exosomes from OA and RA SFB as compared to exosomes from normal SFB. IL-1b stimulation of SFB enhanced the angiogenic activity in their exosomes. However, migration and tube formation were not induced in endothelial cells by cytokine IL-1b alone. miRNA array data showed that 349 miRNAs were changed in OA SFB exosomes as compared to normal SFB expsomes. OA-and RA-SFB exosomes shared 29 miRNAs that were up-regulated and 35 miRNAs that were down-regulated compared normal SFB exosomes. This includes several differentially expressed angiogenesis-related miRNAs. Conclusions: Exosomes from OA SFB accelerate angiogenic activity in HUVECs, and might be involved in OA development by promoting angiogenesis throughout the joint. miRNA array data suggest an OA-related miRNA profile in exosomes that may serve as a novel biomarker. These results support our hypothesis that exosomes containing miRNAs function in a novel regulatory network that contributes to various aspects of OA pathogenesis.

Two classic strategic orientations have been found to pervade the behavior of modern salespersons: a sales orientation (SO) where salespersons use deception or guile to get customers to buy even if they do not need a product, and a customer orientation (CO) where salespersons first attempt to discover the customer's needs and adjust their product and selling approach to meet those needs. Study 1 replicates recent research and finds that the Taq A1 variant of the DRD2 gene is not related to either sales or CO, whereas the 7-repeat variant of the DRD4 gene is related to CO but not SO. Study 2 investigates gene × phenotype explanations of orientation of salespersons, drawing upon recent research in molecular genetics and biological/psychological attachment theory. The findings show that attachment style regulates the effects of DRD2 on CO, such that greater avoidant attachment styles lead to higher CO for persons with the A2/A2 variant but neither the A1/A2 nor A1/A1 variants. Likewise, attachment style regulates the effects of DRD4 on CO, such that greater avoidant attachment styles lead to higher CO for persons with the 7-repeat variant but not other variants. No effects were found on a SO, and secure and anxious attachment styles did not function as moderators.

No abstract

The CRISPR/Cas system has recently emerged as a powerful tool to engineer the genome of an organism. The system is adopted from bacteria where it confers immunity against invading foreign DNA. This work reports the first successful use of the CRISPR/Cas system in Caenorhabditis briggsae (a cousin of the well-known nematode C. elegans), to generate mutations via non-homologous end joining. We recovered deletion alleles of several conserved genes by microinjecting plasmids that express Cas9 endonuclease and an engineered CRISPR RNA corresponding to the DNA sequence to be cleaved. Evidence for somatic mutations and off-target mutations are also reported. Our approach allows for the generation of loss-of-function mutations in C. briggsae genes thereby facilitating a comparative study of gene function.

The synthesis and characterization of iron-sulfur clusters stabilized by dimethylsilyl-bridged cyclopentadienyl groups are reported. The thermal reaction of Me(2)Si(eta(5)-C(5)H(4))(2)Fe(2)(CO)(4) (1) with S(8) yields the tetranuclear cubane-type cluster compound [Me(2)Si(eta(5)-C(5)H(4))(2)](2)Fe(4)S(6) (4) and the pentanuclear cluster compound [Me(2)Si(eta(5)-C(5)H(4))(2)](2)Fe(5)S(12) (3) in high yields. The photochemical reaction of 1 with S(8) yields the tetranuclear cluster compound [Me(2)Si(eta(5)-C(5)H(4))(2)](2)Fe(4)S(6)(CO) (5), which contains one residual terminal carbonyl. The crystal structures of 3 and 4 have been determined. Crystal data: 3.CH(2)Cl(2), monoclinic, C2/c, a = 23.480(13) Å, b = 11.192 (4) Å, c = 17.84 (3) Å, beta = 118.58(9) degrees, V = 4118(7) Å(3), Z = 4, R = 0.078; 4, triclinic, P&onemacr;, a = 8.4787(7) Å, b = 12.9648(9) Å, c = 13.4990(9) Å, alpha = 79.857(8) degrees, beta = 75.293(8) degrees, gamma = 74.041(11) degrees, V = 1370.9(2) Å(3), Z = 2, R = 0.0447. The Fe(5)S(12) core of 3 has a bowtie structure in which a central iron atom is octahedrally coordinated by six sulfur atoms from one tetrasulfido and four disulfido groups. The structure of 4 resembles the structure of the known iron-sulfur cluster Cp(4)Fe(4)S(6). However, 4 shows a markedly enhanced thermal stability compared to Cp(4)Fe(4)S(6). In their cyclic voltammograms, 4 and 5 exhibit electrochemical behavior typical of cubane-type Cp-iron-sulfur clusters, whereas the cyclic voltammogram of 3 is quite different. The nu(CO) mode of 5 has been measured for four different oxidation states of the cluster by means of IR spectroelectrochemical methods. The Mössbauer spectra of 3 and 3(+) are in accordance with their pentanuclear structure.

No abstract

No abstract