IMPORTANCE Posttraumatic stress disorder (PTSD) has been associated in cross-sectional studies with peripheral inflammation. It is not known whether this observed association is the result of PTSD predisposing to inflammation (as sometimes postulated) or to inflammation predisposing to PTSD.OBJECTIVE To determine whether plasma concentration of the inflammatory marker C-reactive protein (CRP) helps predict PTSD symptoms. DESIGN, SETTING, AND PARTICIPANTSThe Marine Resiliency Study, a prospective study of approximately 2600 war zone-deployed Marines, evaluated PTSD symptoms and various physiological and psychological parameters before deployment and at approximately 3 and 6 months following a 7-month deployment. Participants were recruited from 4 all-male infantry battalions imminently deploying to a war zone. Participation was requested of 2978 individuals; 2610 people (87.6%) consented and 2555 (85.8%) were included in the present analysis. Postdeployment data on combat-related trauma were included for 2208 participants (86.4% of the 2555 included) and on PTSD symptoms at 3 and 6 months after deployment for 1861 (72.8%) and 1617 (63.3%) participants, respectively. MAIN OUTCOMES AND MEASURESSeverity of PTSD symptoms 3 months after deployment assessed by the Clinician-Administered PTSD Scale (CAPS). RESULTSWe determined the effects of baseline plasma CRP concentration on postdeployment CAPS using zero-inflated negative binomial regression (ZINBR), a procedure designed for distributions, such as CAPS in this study, that have an excess of zeroes in addition to being positively skewed. Adjusting for the baseline CAPS score, trauma exposure, and other relevant covariates, we found baseline plasma CRP concentration to be a highly significant overall predictor of postdeployment CAPS scores (P = .002): each 10-fold increment in CRP concentration was associated with an odds ratio of nonzero outcome (presence vs absence of any PTSD symptoms) of 1.51 (95% CI, 1.15-1.97; P = .003) and a fold increase in outcome with a nonzero value (extent of symptoms when present) of 1.06 (95% CI, 0.99-1.14; P = .09). CONCLUSIONS AND RELEVANCEA marker of peripheral inflammation, plasma CRP may be prospectively associated with PTSD symptom emergence, suggesting that inflammation may predispose to PTSD.

Herpes simplex virus type 1 encodes a heterotrimeric helicase-primase complex that is composed of the products of the UL5, UL52, and UL8 genes. A subcomplex consisting of the UL5 and UL52 proteins retains all the enzymatic activities exhibited by the holoenzyme in vitro. The UL52 protein contains a putative zinc finger at its C terminus which is highly conserved among both prokaryotic and eukaryotic primases. We constructed a mutation in which two highly conserved cysteine residues in the zinc finger motif were replaced with alanine residues. A UL52 expression plasmid containing the mutation in the zinc finger region is unable to support the growth of a UL52 mutant virus in a transient complementation assay. Wild type and mutant UL5⅐UL52 subcomplexes were purified from insect cells infected with recombinant baculoviruses. Surprisingly, the mutant protein was severely affected in all biochemical activities tested; no helicase or primase activities could be detected, and the mutant protein retains only about 9% of wild type levels of single-stranded DNA-dependent ATPase activity. Gel mobility shift assays showed that DNA binding is severely affected as well; the mutant subcomplex only retains approximately 8% of wild type levels of binding to a forked substrate. On the other hand, the mutant protein retains its ability to interact with UL5 as indicated by copurification and with UL8 as indicated by a supershifted band in the gel mobility shift assay. In addition, the ability of individual subunits to bind single-stranded DNA was examined by photo crosslinking. In the wild type UL5⅐UL52 subcomplex, both subunits are able to bind an 18-mer of oligo(dT). The mutant subcomplex was severely compromised in the ability of both UL5 and UL52 to bind the oligonucleotide; total cross-linking was only 2% of wild type levels. These results are consistent with the proposal that the putative zinc binding motif of UL52 is required not only for binding of the UL52 subunit to DNA and for primase activity but also for optimal binding of UL5 to DNA and for the subsequent ATPase and helicase activities.Herpes simplex virus type 1 (HSV-1) 1 encodes a heterotrimeric DNA helicase-primase complex composed of the products of the UL5, UL52, and UL8 genes (1, 2). All three genes are essential for viral DNA replication (3-7). This protein complex has been shown to possess ssDNA-dependent NTPase, 5Ј to 3Ј DNA helicase, and DNA primase activities (1, 2, 8 -10). The HSV-1 helicase-primase complex can be isolated from insect cells that have been simultaneously infected with recombinant baculoviruses that express each of the three subunits (8). A subassembly consisting of the UL5 and UL52 gene products also exhibits all the enzymatic activities of the holoenzyme (11). The UL5 protein contains seven conserved motifs found in all members of helicase Superfamily I which comprises DNA and RNA helicases from bacteria, viruses, and eukaryotes (12). Mutations in conserved residues in the helicase motifs have been shown to abolish the ability of mutant UL...

Background: Mice lacking the neurosecretory protein Chromogranin A are obese, presumably because of resistance to catecholamines and leptin. Results: Catestatin (CST) reduces adiposity, an effect likely mediated by restoring leptin sensitivity and modulating adrenergic signaling. Conclusion: CST promotes lipolysis by blocking ␣-AR signaling and stimulating fatty acid oxidation. Significance: We propose CST as a candidate antiobesity agent.

Previous studies have shown that infection of G 0 -synchronized human fibroblasts by human cytomegalovirus (HCMV) results in a block to cellular DNA synthesis. In this study, we have examined the effect of viral infection on the formation of the host cell DNA prereplication complex (pre-RC). We found that the Cdc6 protein level was significantly upregulated in the virus-infected cells and that there was a delay in the expression of the Mcm family of proteins. The loading of the Mcm proteins onto the DNA pre-RC complex also appeared to be defective in the virus-infected cells. This inhibition of DNA replication licensing was associated with the accumulation of geminin, a replication inhibitor. Cdt1, which participates in the loading of the Mcm proteins, was also downregulated and modified differentially in the infected cells. Early viral gene expression was sufficient for the virus-induced alteration of the pre-RC, and the immediate-early protein IE1 was not required. These studies show that the inhibition of replication licensing in HCMV-infected cells is one of the multiple pathways by which the virus dysregulates the host cell cycle.Human cytomegalovirus (HCMV), an ubiquitous betaherpesvirus, is the leading viral cause of birth defects and poses a serious health threat to immunocompromised individuals (40). The development of strategies to prevent HCMV infection requires an understanding of the initial interactions between the virus and the host cell that promote the progression of the viral replication cycle and subsequent pathogenesis. As is the case with mitogens, HCMV infection of quiescent cells results in the rapid activation of the cellular proto-oncogenes c-fos, c-jun, and c-myc as well as an increased expression of ornithine decarboxylase, thymidine kinase, DNA polymerase alpha, and dihydrofolate reductase (1,8,19,22,52). In addition, increased levels of p53 and hyperphosphorylated Rb are observed in the virus-infected cells (23). HCMV also induces elevated levels of cyclin E and cyclin B and their associated kinase activities (23). In contrast, the expression of cyclin A and its associated kinase activity is inhibited (23). These combined effects suggest that HCMV adopts a strategy of early cellular activation that facilitates viral replication but simultaneously inhibits host cell DNA synthesis by an undefined mechanism.Work from our laboratory and others has shown that the HCMV infection blocks cell cycle progression in primary human fibroblasts. In these studies, cells that were synchronized by serum starvation, contact inhibition, or both conditions, as well as asynchronous, proliferating cells, were used (2,7,23,29,46). The arrest occurs primarily at G 1 /S, but blockage at other points in the cell cycle also has been observed. It has been proposed that the immediate-early proteins (IE1 and IE2) and the virion constituent UL69 of HCMV contribute to the virus-mediated alteration in cell growth control (3,30,36,55,56).Previously, it was demonstrated that the cell cycle phase at the time of the inf...

Chromogranin A (CHGA/Chga), a proprotein, widely distributed in endocrine and neuroendocrine tissues (not expressed in muscle, liver, and adipose tissues), generates at least four bioactive peptides. One of those peptides, pancreastatin (PST), has been reported to interfere with insulin action. We generated a Chga knock-out (KO) mouse by the targeted deletion of the Chga gene in neuroendocrine tissues. KO mice displayed hypertension, higher plasma catecholamine, and adipokine levels and lower IL-6 and lipid levels compared with wild type mice. Liver glycogen content was elevated, but the nitric oxide (NO) level was diminished. Glucose, insulin, and pyruvate tolerance tests and hyperinsulinemic-euglycemic clamp studies established increased insulin sensitivity in liver but decreased glucose disposal in muscle. Despite higher catecholamine and ketone body levels and muscle insulin resistance, KO mice maintained euglycemia due to increased liver insulin sensitivity. Suppressed mRNA abundance of phosphoenolpyruvate carboxykinase and glucose-6-phosphatase (G6Pase) in KO mice further support this conclusion. PST administration in KO mice stimulated phosphoenolpyruvate carboxykinase and G6Pase mRNA abundance and raised the blood glucose level. In liver cells transfected with G6Pase promoter, PST caused transcriptional activation in a protein kinase C (PKC)-and NO synthase-dependent manner. Thus, PST action may be mediated by suppressing IRS1/ 2-phosphatidylinositol 3-kinase-Akt-FOXO-1 signaling and insulin-induced maturation of SREBP1c by PKC and a high level of NO. The combined effects of conventional PKC and endothelial NO synthase activation by PST can suppress insulin signaling. The rise in blood PST level with age and in diabetes suggests that PST is a negative regulator of insulin sensitivity and glucose homeostasis.Chromogranin A (CHGA/Chga), 2 the index member of the chromogranin/secretogranin protein family (1, 2), is a proprotein that gives rise to biologically active peptides such as the dysglycemic hormone pancreastatin (PST; human CHGA-(250 -301)) (3), the vascular smooth muscle vasodilator vasostatin (human CHGA-(1-76)) (4), and a catecholamine release inhibitory peptide, catestatin (human CHGA-(352-372); bovine Chga-(344 -364)) (5). Several studies, including in vivo analyses of experimental animals (6, 7), showed that PST inhibits insulin release in response to glucose (3), reduces glucose uptake in adipocytes and hepatocytes (8), and triggers glycogenolysis (6). Genetic analysis in humans supports a role for PST in decreasing glucose uptake by ϳ50% (9) while increasing the spillover of free fatty acids but not amino acids. Moreover the PST level is elevated in patients with Type 2 diabetes mellitus (9 -11). Taken together, the data suggest that PST is an important player in metabolism.To further delineate the role of PST in metabolism, we tested whether removal of PST, a negative regulator of insulin action, stabilizes glucose levels in knock-out (KO) mice and protects against metabolic disorders. To t...