Drug-dependent neural plasticity related to drug addiction and schizophrenia can be modeled in animals as behavioral sensitization, which is induced by repeated noncontingent or self-administration of many drugs of abuse. Molecular mechanisms that are critical for behavioral sensitization have yet to be specified. Long-term depression (LTD) of alpha-amino-3-hydroxy-5-methyl-isoxazole-4-propionic acid receptor (AMPAR)-mediated synaptic transmission in the brain has been proposed as a cellular substrate for learning and memory. The expression of LTD in the nucleus accumbens (NAc) required clathrin-dependent endocytosis of postsynaptic AMPARs. NAc LTD was blocked by a dynamin-derived peptide that inhibited clathrin-mediated endocytosis or by a GluR2-derived peptide that blocked regulated AMPAR endocytosis. Systemic or intra-NAc infusion of the membrane-permeable GluR2 peptide prevented the expression of amphetamine-induced behavioral sensitization in the rat.

BackgroundFinancial relationships between organizations that produce clinical practice guidelines and biomedical companies are vulnerable to conflicts of interest. We sought to determine whether organizations that produce clinical practice guidelines have financial relationships with biomedical companies and whether there are associations between organizations’ conflict of interest policies and recommendations and disclosures provided in guidelines.Methods and FindingsWe conducted a cross-sectional survey and review of websites of 95 national/international medical organizations that produced 290 clinical practice guidelines published on the National Guideline Clearinghouse website from January 1 to December 31, 2012. Survey responses were available for 68% (65/95) of organizations (167/290 guidelines, 58%), and websites were reviewed for 100% (95/95) of organizations (290/290 guidelines, 100%). In all, 63% (60/95) of organizations producing clinical practice guidelines reported receiving funds from a biomedical company; 80% (76/95) of organizations reported having a policy for managing conflicts of interest. Disclosure statements (disclosing presence or absence of financial relationships with biomedical companies) were available in 65% (188/290) of clinical practice guidelines for direct funding sources to produce the guideline, 51% (147/290) for financial relationships of the guideline committee members, and 1% (4/290) for financial relationships of the organizations producing the guidelines. Among all guidelines, 6% (18/290) disclosed direct funding by biomedical companies, 40% (117/290) disclosed financial relationships between committee members and biomedical companies (38% of guideline committee members, 773/2,043), and 1% (4/290) disclosed financial relationships between the organizations producing the guidelines and biomedical companies. In the survey responses, 60 organizations reported the procedures that they included in their conflict of interest policies (158 guidelines): guidelines produced by organizations reporting more comprehensive conflict of interest policies (per additional procedure, range 5–17) included fewer positive (rate ratio [RR] 0.91, 95% CI 0.86–0.95) and more negative (RR 1.32, 95% CI 1.09–1.60) recommendations regarding patented biomedical products. The clinical practice guidelines produced by organizations reporting more comprehensive conflict of interest policies were also more likely to include disclosure statements for direct funding sources (odds ratio [OR] 1.31, 95% CI 1.10–1.56) and financial relationships of guideline committee members (OR 1.36, 95% CI 1.09–1.79), but not financial relationships of the organizations (0 disclosures). Limitations of the study include the use of the National Guideline Clearinghouse as the single source of clinical practice guidelines and the self-report of survey responses and organizations’ website postings.ConclusionsFinancial relationships between organizations that produce clinical practice guidelines and biomedical companies are common and...

Burkholderia cepacia complex is a life-threatening group of pathogens for patients with chronic granulomatous disease (CGD), whose phagocytes are unable to produce reactive oxygen species (ROS). Unlike other CGD pathogens, B. cepacia complex is particularly virulent, characteristically causing septicemia, and is the bacterial species responsible for most fatalities in these patients. We found that a nonmucoid Burkholderia cenocepacia (a predominant species in the B. cepacia complex) isolate was readily ingested by normal human neutrophils under nonopsonic conditions and promoted apoptosis in these cells. The proapoptotic effect was not due to secreted bacterial products, but was dependent on bacterial viability. Phagocytosis was associated with a robust production of ROS, and the apoptotic neutrophils could be effectively cleared by monocyte-derived macrophages. The proapoptotic effect of B. cenocepacia was independent of ROS production because neutrophils from CGD patients were rendered apoptotic to a similar degree as control cells after challenge. More importantly, neutrophils from CGD patients, but not from normal individuals, were rendered necrotic after phagocytosis of B. cenocepacia. The extreme virulence of B. cepacia complex bacteria in CGD, but not in immunocompetent hosts, could be due to its necrotic potential in the absence of ROS.

Innate immunodeficiency has recently been reported resulting from the Q293X IRAK-4 mutation, with consequent defective TLR/IL-1R signalling. Here we report a method for the rapid allele-specific detection of this mutation and demonstrate both cell-type specificity and ligand specificity in defective IRAK-4-deficient cellular responses, indicating differential roles for this protein in human peripheral blood mononuclear cells and primary dermal fibroblasts, and in LPS, IL-1β and TNF-α signalling. We demonstrate transcriptional and post-transcriptional defects, despite NF-κB signalling and intact MyD88-independent signalling, and propose that dysfunctional Complex 1 (IRAK1/TRAF6/TAK1) signalling, as a consequence of IRAK-4-deficiency, generates specific defects in mitogen-activated protein kinase activation that could underpin this patient’s innate immunodeficiency. These studies demonstrate the importance of studying primary human cells bearing a clinically relevant mutation; they underscore the complexity of innate immune signalling and illuminate novel roles for IRAK-4 and the fundamental importance of accessory pro-inflammatory signalling to normal human innate immune responses and immunodeficiencies.

A proportion of individuals vaccinated with live attenuated Oka varicella-zoster virus (VZV) vaccine subsequently develop attenuated chicken pox and/or herpes zoster. To determine whether postvaccination varicella infections are caused by vaccine or wild-type virus, a simple method for distinguishing the vaccine strain from wild-type virus is required. We have developed a TaqMan real-time PCR assay to detect and differentiate wild-type virus from Oka vaccine strains of VZV. The assay utilized two fluorogenic, minor groove binding probes targeted to a single nucleotide polymorphism in open reading frame 62 that distinguishes the Oka vaccine from wild-type strains. VZV DNA could be genotyped and quantified within minutes of thermocycling completion due to real-time monitoring of PCR product formation and allelic discrimination analysis. The allelic discrimination assay was performed in parallel with two standard PCR-restriction fragment length polymorphism (RFLP) methods on 136 clinical and laboratory VZV strains from Canada, Australia, and Japan. The TaqMan assay exhibited a genotyping accuracy of 100% and, when compared to both PCR-RFLP methods, was 100 times more sensitive. In addition, the method was technically simpler and more rapid. The TaqMan assay also allows for high-throughput genotyping, making it ideal for epidemiologic study of the live attenuated varicella vaccine. Varicella-zoster virus (VZV) is the etiological agent of varicella chicken pox and herpes zoster (HZ). Both varicella and HZ are self-limiting infections in healthy individuals but may cause more severe disease in immunocompromised hosts (2,8,23,38).A live attenuated Oka vaccine (V-Oka) strain was created in 1974 by the repeated passage of a Japanese clinical isolate in tissue culture (34). The vaccine was licensed in the United States in 1995 (2, 4) and in Canada in 1998 (27) and is currently used in many other countries (3, 40). Extensive clinical trials have shown the vaccine to be highly effective in both healthy and immunocompromised individuals (11,19,30,33). However, a low incidence of superinfection with wild-type (WT) strains after immunization, termed breakthrough varicella, has been well documented (3,10,21,39). Vaccine-associated rash occurs in 4 to 7% of healthy individuals (4, 40) and in up to 40% of leukemic vaccinees (10, 11). The vaccine strain does establish neuronal latency, and hence, reactivation of vaccine virus in and transmission from healthy and immunocompromised individuals is possible (11,20,21). As the number of vaccinees is increasing worldwide (31), the clinical management of HZ is becoming more complex. Given the fact that the vaccine virus is less pathogenic (39) and also less transmissible than WT virus (37), a timely differentiation of vaccine or WT disease would be clinically useful in situations with pregnant or immunocompromised household or hospital contacts, where prompt postexposure treatment may be indicated.The original method used to differentiate WT and vaccine VZV was restriction fragment leng...