Biochemical anti-opioid action of NPFF2 receptors in rat spinal cord

Neuropeptide FF (NPFF) agonists counteract the cellular opioid actions. We demonstrate for the first time a biochemical anti-opioid effect of NPFF receptors in the rat spinal cord by using the [(35)S]GTPgammaS binding assay in autoradiography. The mu agonist DAMGO as well as the potent and selective NPFF(2) agonist dNPA, stimulated [(35)S]GTPgammaS binding at different optimal GDP concentrations. dNPA decreased the effects induced by DAMGO alone; the maximal of G-protein coupling was decreased but not the potency of opioid agonist. We conclude that NPFF(2) receptors are coupled to G-protein in the rat spinal cord and could exert a molecular anti-opioid effect.     

Dolosigranulum pigrum causing nosocomial pneumonia and septicemia

We report a case of non-ventilator-associated nosocomial pneumonia and septicemia due to Dolosigranulum pigrum, a rare gram-positive opportunistic pathogen. The organism was isolated from bronchoalveolar lavage fluid and blood of a debilitated patient. D. pigrum was identified after 16S rRNA gene sequencing.     

Significant genetic and antigenic variability within the env gene of systemic human immunodeficiency virus type 1 group O populations during the natural course of a heterosexual infection: a pilot study

We assessed the genetic and the antigenic variability within the env gene of peripheral blood human immunodeficiency virus (HIV) type 1 (HIV-1) group O populations during the natural course of a female heterosexual infection. Our data revealed the existence of a significant increase in amino acid sequence variability within the C2-V3 and gp41 regions (P = 0.023 and P < 0.001, respectively) in association with substitutions within neutralizing epitope sequences usually selected for HIV serological assays. These antigenic variations might significantly decrease the sensitivity of classical HIV enzyme-linked immunosorbent assays with blood samples of subjects heterosexually infected by HIV-1 group O strains. These findings may be of significant use both to devise diagnostic tools and to pursue suitable therapeutic modalities in cases of heterosexual infection by outlier HIV-1 strains.     

Design of an endoscopic optical reference to be used for autofluorescence bronchoscopy with a commercially available diagnostic autofluorescence endoscopy (DAFE) system

We present the design of a sterilizable optical reference to characterize and quantify the inter-patient variations in tissue autofluorescence during autofluorescence bronchoscopy with Richard Wolf's diagnostic autofluorescence endoscopy (DAFE) system. The reference was designed to have optical and spectral properties similar to those of the human bronchial wall in spectral conditions corresponding to autofluorescence bronchoscopy conducted with the DAFE system (fluorescence excitation at 390-470 nm and red backscattering light at 590-680 nm). The reference's effective attenuation coefficient and reflectance were measured at 675 nm. In addition, its fluorescence emission spectrum was determined under 430 nm wavelength excitation. The reference is photostable, reproducible, biocompatible and small enough to be easily inserted through the working channel of a conventional bronchofibrescope. This cylindrical (length: 2 mm; diameter: 2 mm) optical reference was validated in a clinical environment.     

Protective DNA vaccination against myelin oligodendrocyte glycoprotein is overcome by C3d in experimental autoimmune encephalomyelitis

Complement receptor 2 (CR2) and its physiological ligand, C3d, known for its molecular adjuvant property on the immune response, exhibit opposite effects with regard to autoimmunity. Although CR2 has been implicated in maintaining self-tolerance, recent studies reported a role for C3d signaling to CR2 in tolerance breakdown to self-antigens and the initiation of inflammatory autoimmune pathologies. In the present study, we have investigated the effect of C3d in a model of tolerogenic DNA vaccination encoding the myelin oligodendrocyte glycoprotein (MOG-DNA) which protected mice from the induction of an experimental autoimmune encephalomyelitis (EAE). We show that fusing two or three copies of C3d to MOG overcomes the protective effect of DNA vaccination. Multimeric C3d was able to revert the unresponsiveness state of specific T cells induced by MOG-DNA, independently of a modification in the Th1/Th2 cytokine pattern. Interestingly, the adjuvant effect of C3d was not sufficient to boost the anti-MOG antibody response after DNA vaccination. These findings suggest that C3d might be involved in self-tolerance breakdown and could contribute to the pathogenesis of central nervous system autoimmune disorders.     

Generation of llama single-domain antibodies against methotrexate, a prototypical hapten

Single-domain antibodies specific to methotrexate (MTX) were obtained after immunization of one llama (Llama glama). Specific VHH domains (V-D-J-REGION) were selected by panning from an immune-llama library using phage display technology. The antibody fragments specific to MTX were purified from Escherichia coli (C41 strain) periplasm by immobilized metal affinity chromatography with an expression level of around 10mg/L. A single band around 16,000Da corresponding to VHH fragments was found after analysis by SDS-PAGE and Western blotting, while competition ELISA demonstrated selective binding to soluble MTX. Surface plasmon resonance (SPR) analysis showed that anti-MTX VHH domains had affinities in the nanomolar range (29-515nM) to MTX-serum albumin conjugates. The genes encoding anti-MTX VHH were found by IMGT/V-QUEST to be similar to the previously reported llama and human IGHV germline genes. The V-D and D-J junction rearrangements in the seven anti-MTX CDR3 sequences indicate that they were originated from three distinct progenitor B cells. Our results demonstrate that camelid single-domain antibodies are capable of high affinity binding to low molecular weight hydrosoluble haptens. Furthermore, these anti-MTX VHH give new insights on how the antigen binding repertoire of llama single-domain antibody can provide combining sites to haptens in the absence of a VL. This type of single-domain antibodies offers advantages compared to murine recombinant antibodies in terms of production rate and sequence similarity to the human IGHV3 subgroup genes.     

Free and intracellular bacteria on peripheral blood smears: an uncommon situation related to an adverse prognosis

Bacterial infections are responsible for several changes in the cell blood count, which are usually non specific, although some morphological changes of polymorphonuclear neutrophils may be indicative of sepsis. The presence of bacteria on peripheral blood smears is a rare but extreme situation, related in most instances to a fatal prognosis. The presence of both free and intracellular bacteria was observed in the peripheral blood smear of a critically ill patient with a pneumococcal septicaemia which led to a fatal outcome within the next following hours. If the finding of bacteria on the blood smear is a sign of severe sepsis, the literature review shows that less than 10% of septic patients demonstrate bacteria on the blood smear, and routine search for the diagnosis of sepsis is not recommended. Samples taken from infected central venous catheters are another situation of bacteraemia which must be known, but prognosis is usually not fatal if prompt medical care is performed. Some preanalytical conditions are also associated with the presence of bacteria on the peripheral blood smear, but unrelated to infection of the relevant patient.     

Drug-induced agranulocytosis in rheumatology. A retrospective study of 12 cases

OBJECTIVE: the aim of this study is to determine the clinical characteristics of drug-induced agranulocytosis in rheumatology. PATIENTS AND METHOD: it is a retrospective monocentric study, including all cases of rheumatologic drug-induced agranulocytosis followed between January 1985 and December 2005. RESULTS: eight female and 4 men, mean age 62.5 years (range: 17-79), were included in the present study. The causative drugs were: anti-inflammatory agents (n=5, 41.6%), methotrexate (n=3, 25%), sulfasalazine (n=2, 16.6%), noramidopyrine (n=1) and dapsone (n=1). Main clinical features included infectious disorders in 9 cases (75%) with 4 cases of septicemia (33.3%) and 2 cases of septic shock (16.6%). The mean neutrophils count was 0.132 x 10(9)/L (range, 0-0.4). Outcome was favorable in 11 patients (91.6%). One patient died of septic complications. CONCLUSION: in rheumatology, drug-induced agranulocytosis is a rare but life-threatening disorder.