Molluscum contagiosum virus (MCV) lesions from Spanish human immunodeficiency virus (HIV)-negative patients were clinically examined and analyzed for virus detection and typing. In a study of 147 patients, 97 (66%) were children under 10 years, of whom 49% had atopic dermatitis. MCV lesions were morphologically indistinguishable among the different age groups, but atopic patients presented larger lesions compared with patients without the disorder. In adults, lesions were observed mainly on the genitals. MCVI was the predominant subtype. The deduced MCVI/MCVII ratio (146:1) was much higher than that found in other geographical areas. Protein preparations of the virus-induced lesions were immunoblotted with sera from 25 MCVI patients. The host-serum antibody response was weak and variable, although no significant differences were found between atopic and nonatopic patients. Three immunoreactive proteins of 74/80, 60, and 35 kDa were detected in almost all the analyzed sera. The 35 and 74/80-kDa proteins were virus specific, whereas the 60-kDa protein band was composed of a mix of human keratins. Immunoblotting of MCV lesions and vaccinia virus-infected cell extracts with either MCV patient serum or a rabbit antiserum against vaccinia virus showed no cross-reactivity of these two human poxviruses at the antigenic level.

Neuroblastoma cell lines (SK-N-SH and SK-N-MC) were induced to differentiate, as detected by the expression of neurofilament proteins of 68 and 200 kDa, and to express adhesion molecules (intercellular adhesion molecule-1 and vascular cell adhesion molecule) after stimulation with tumour necrosis factor-alpha (TNF-alpha). This induction was accompanied by the arrest of cell growth. The induction of neuroblastoma adhesion by TNF-alpha could be inhibited by the nitric oxide synthase inhibitors, L-N-monomethyl arginine (L-NMMA) and L-N6-(1-imidoethyl)-lysine (highly specific for the inducible enzyme), but not by the inactive enantiomer D-NMMA. These results indicate that TNF-alpha induces the adhesion of neuroblastoma cells via nitric oxide. This was confirmed by the finding that the adhesion/differentiation of SK-N-SH and SK-N-MC cells can be directly induced by the addition of nitric oxide donors, sodium nitroprusside and S-nitroso-N-acetyl-penicillamine, into the culture medium. The isoform of the nitric oxide synthase induced in human neuroblastoma cells by TNF-alpha treatment was identified enzymatically as isoform II by Western blotting and by the polymerase chain reaction. Thus TNF-alpha induces the in vitro adhesion/differentiation of human neuroblastoma cells through nitric oxide synthesized by a calcium-independent inducible form of nitric oxide synthase, clearly indicating that isoform II of nitric oxide synthase can be expressed in human neuronal cell types.

Addition of nitric oxide (NO) donors to mitogen-activated human immunodeficiency virus type 1 (HIV-1

CD81 is expressed on lymphocytes and confers HCV viral infectivity support. The aim of our study was to quantify CD81 expression in peripheral blood B-and T-cells of HCV/HIV-coinfected patients and healthy subjects to examine its association with several HCV virological characteristics and the therapeutic responsiveness to HCV antiviral treatment. MethodsWe carried out a cross-sectional study on 122 naïve patients. For a duration of 48 weeks, 24 out of 122 patients underwent HCV antiviral therapy with interferon (IFN)-a and ribavirin. T-and B-cell subsets were analysed by flow cytometry. ResultsWe found that HIV/HCV coinfected patients with HCV-RNA ! 850 000 IU/mL had lower values of %CD19 1 CD81-CD62L 1 and %CD19 1 CD62L 1 ; and higher values of CD19 1 CD81 1 CD62L À and CD19 1 CD81 1 percentages and absolute counts than patients with HCV-RNA o850 000 IU/mL. Similarly, HIV/HCV coinfected patients with the genotype 1 had lower values of %CD19 1 CD81 À CD62L 1 and higher values of CD3 1 CD81 1 CD62L À and CD3 1 CD81 1 percentages and absolute counts than patients without genotype 1. Moreover, we found that HIV/HCV coinfected patients had higher values of %CD19 1 HLA-DR 1 CD25 1 , %CD19 1 CD40 1 CD25 1 and %CD19 1 CD25 1 than healthy control patients. When we studied the B-and T-cell subset kinetics of 24 HIV/HCV coinfected patients on HCV antiviral therapy, we found a significant decrease in CD3 1 CD81 1 and CD3 1 CD81 1 CD62L À subsets and a significant increase in CD3 1 CD62L 1 and CD3 1 CD81 1 CD62L 1 percentages and absolute counts, but the variation in these markers disappeared several months after stopping the treatment. ConclusionsWe observed a different pattern of CD81 T-cell and B-cell levels in naïve HIV/HCV coinfected patients according to HCV virological status and their subsequent variations during HCV antiviral treatment. CD81 expression might influence HCV pathogenesis and response to HCV antiviral treatment.

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Two forms of the regulatory subunit of the type II cAMP-dependent protein kinase (RII55 and RII52) were identified from bovine heart by gel electrophoretic behaviour. After autophosphorylation the RII55 isoform migrated more slowly (RII55/57) while the migration of RII52 isoform did not shift. Both isoforms showed different affinity for cAMP. The RII55/57 isoform was eluted from a cAMP-agarose column at 10 mM cAMP at low ionic strength whereas the RII52 isoform required cAMP, plus 2M NaCl. Partial proteolysis, using trypsin or formic acid, of autophosphorylated regulatory subunit isoforms resulted in different cleavage pattern as determined by peptide mapping. However, the V8 125I-peptides patterns of both isoforms are quite similar. Incubation of partially purified holoenzyme with 10 nM [gamma-32P]ATP (low ATP concentration) yielded a single band of Mr = 57,000 which corresponds to the RII55/57 isoform. The incubation, however, at 20 microM [gamma-32P]ATP yielded two phosphobands corresponding to both RII55/57 and RII52 isoforms. The phosphorylation of RII52 took place with a lower efficiency and was more sensitive to the cAMP than the corresponding phosphorylation of the RII55/57.