Role of peptidergic neurons in ocular herpes simplex infection in mice

Peptide-containing nerve fibers (peptidergic fibers) abundantly innervate the mammalian cornea. We investigated their role in ocular herpes simplex infection in mice by using capsaicin, which causes degeneration and permanent loss of peptidergic neurons in neonates and temporary peptide depletion in adult animals. The corneas of neonatally denervated BALB/c mice were observed for capsaicin-induced keratitis at 11-14 wk of age and were then infected bilaterally with herpes simplex virus 1 (HSV-1); trigeminal (TG) ganglia were cocultivated 6 wk later to establish the rate of latent infection. We also applied capsaicin eye drops to adult BALB/c mice that had been infected with HSV-1 6 wk earlier, and measured viral shedding before, and 3 days and 2 months after, administration of capsaicin drops; TG ganglia of these animals were cocultivated at 3 days and 2 months after capsaicin application. Neurotrophic keratitis was found in 81% of neonatally denervated animals; mortality rate due to HSV-1 infection was reduced from 80% in the controls to 24% in the capsaicin-treated group, and recovery of latent virus by cocultivation was reduced by 50%. Viral shedding could not be produced by capsaicin eye drops in adult animals infected with HSV-1. However, recovery of latent virus was significantly reduced in TG ganglia sampled 3 days and 2 months after capsaicin drops were instilled. Our findings suggest 1) that peptidergic fibers play a crucial role in the establishment of trigeminal HSV-1 latency and 2) that reactivation of latently infected ganglia can be inhibited by topical capsaicin.     

Scheduling parallel applications in distributed networks

Prophet is a run-time scheduling system designed to support the efficient execution of parallel applications written in the Mentat programming language (Grimshaw, 1993). Prior results demonstrated that SPMD applications could be scheduled automatically in an Ethernet-based local-area workstation network with good performance (Weissman and Grimshaw, 1994 and 1995). This paper describes our recent efforts to extend Prophet along several dimensions: improved overhead control, greater resource sharing, greater resource heterogeneity, wide-area scheduling, and new application types. We show that both SPMD and task parallel applications can be scheduled effectively in a shared heterogeneous LAN environment containing ethernet and ATM networks by exploiting the application structure and dynamic run-time information. This revised version was published online in July 2006 with corrections to the Cover Date.     

Ubiquitin-proteasome-dependent degradation of a mitofusin, a critical regulator of mitochondrial fusion

The mitochondrion is a dynamic membranous network whose morphology is conditioned by the equilibrium between ongoing fusion and fission of mitochondrial membranes. In the budding yeast, Saccharomyces cerevisiae, the transmembrane GTPase Fzo1p controls fusion of mitochondrial outer membranes. Deletion or overexpression of Fzo1p have both been shown to alter the mitochondrial fusion process indicating that maintenance of steady-state levels of Fzo1p are required for efficient mitochondrial fusion. Cellular levels of Fzo1p are regulated through degradation of Fzo1p by the F-box protein Mdm30p. How Mdm30p promotes degradation of Fzo1p is currently unknown. We have now determined that during vegetative growth Mdm30p mediates ubiquitylation of Fzo1p and that degradation of Fzo1p is an ubiquitin-proteasome-dependent process. In vivo, Mdm30p associates through its F-box motif with other core components of Skp1-Cullin-F-box (SCF) ubiquitin ligases. We show that the resulting SCF(Mdm30p) ligase promotes ubiquitylation of Fzo1p at mitochondria and its subsequent degradation by the 26S proteasome. These results provide the first demonstration that a cytosolic ubiquitin ligase targets a critical regulatory molecule at the mitochondrial outer membrane. This study provides a framework for developing an understanding of the function of Mdm30p-mediated Fzo1p degradation in the multistep process of mitochondrial fusion.     

Azathioprine's inhibitory effect on prostaglandin E2 production is not via cyclooxygenase inhibition

Azathioprine, a widely used antimetabolite, is also known for its anti-inflammatory action in rheumatic disorders and in uveitis, an inflammation of the eye, both of which are associated with increased production of prostaglandin E2. Recently we demonstrated that prostaglandin E2 production by rabbit retina/choroid was inhibited by azathioprine and suggested that this inhibitory effect may underlie the drug's antiinflammatory action. In the present study we showed that azathioprine's inhibition of prostaglandin E2 synthesis by the rat retina/choroid was reversed by addition of arachidonic acid, indicating that inhibition occurred through lack of availability of arachidonic acid, similar to the mechanism underlying the inhibitory effect of the steroidal anti-inflammatory drugs on prostaglandin E2. This study rules out the possibility that azathioprine's suppressive activity is via inhibition of the cyclooxygenase pathway.     

Novel DNA mismatch-repair activity involving YB-1 in human mitochondria

Maintenance of the mitochondrial genome (mtDNA) is essential for proper cellular function. The accumulation of damage and mutations in the mtDNA leads to diseases, cancer, and aging. Mammalian mitochondria have proficient base excision repair, but the existence of other DNA repair pathways is still unclear. Deficiencies in DNA mismatch repair (MMR), which corrects base mismatches and small loops, are associated with DNA microsatellite instability, accumulation of mutations, and cancer. MMR proteins have been identified in yeast and coral mitochondria; however, MMR proteins and function have not yet been detected in human mitochondria. Here we show that human mitochondria have a robust mismatch-repair activity, which is distinct from nuclear MMR. Key nuclear MMR factors were not detected in mitochondria, and similar mismatch-binding activity was observed in mitochondrial extracts from cells lacking MSH2, suggesting distinctive pathways for nuclear and mitochondrial MMR. We identified the repair factor YB-1 as a key candidate for a mitochondrial mismatch-binding protein. This protein localizes to mitochondria in human cells, and contributes significantly to the mismatch-binding and mismatch-repair activity detected in HeLa mitochondrial extracts, which are significantly decreased when the intracellular levels of YB-1 are diminished. Moreover, YB-1 depletion in cells increases mitochondrial DNA mutagenesis. Our results show that human mitochondria contain a functional MMR repair pathway in which YB-1 participates, likely in the mismatch-binding and recognition steps.