Panbacterial real-time PCR to evaluate bacterial burden in chronic wounds treated with Cutimed™ Sorbact™

The impact of polymicrobial bacterial infection on chronic wounds has been studied extensively, but standard bacteriological analysis is not always sensitive enough. Molecular approaches represent a promising alternative to the standard bacteriological analysis. This work aimed to assess the usefulness of a panbacterial quantitative real-time PCR reaction to quantitate the total bacterial load in chronic wounds treated with Cutimed™ Sorbact™, a novel therapeutic approach based on hydrophobic binding of bacteria to a membrane. The results obtained by panbacterial real-time PCR on conserved sequences of the bacterial 16S gene show that the bacterial burden significantly decreased in 10 out of 15 healing chronic wounds, and did not change in 5 out of 5 non-healing chronic wounds. On the contrary, classical culture for S. aureus and P. aeruginosa, and real-time PCR for Bacteroides and Fusobacterium did not show any correlation with the clinical outcome. Our study also shows that quantification of chronic wounds by panbacterial real-time PCR is to be performed on biopsies and not on swabs. These results show that panbacterial real-time PCR is a promising and quick method of determining the total bacterial load in chronic wounds, and suggest that it might be an important biomarker for the prognosis of chronic wounds under treatment

Serum IgG against Simian Virus 40 antigens are hampered by high levels of sHLA-G in patients affected by inflammatory neurological diseases, as multiple sclerosis

Background: Many investigators detected the simian polyomavirus SV40 footprints in human brain tumors and neurologic diseases and recently it has been indicated that SV40 seems to be associated with multiple sclerosis (MS) disease. Interestingly, SV40 interacts with human leukocyte antigen (HLA) class I molecules for cell entry. HLA class I antigens, in particular non-classical HLA-G molecules, characterized by an immune-regulatory function, are involved in MS disease, and the levels of these molecules are modified according with the disease status. Objective: We investigated in serum samples, from Italian patients affected by MS, other inflammatory diseases (OIND), non-inflammatory neurological diseases (NIND) and healthy subjects (HS), SV40-antibody and soluble sHLA-G and the association between SV40-prevalence and sHLA-G levels. Methods: ELISA tests were used for SV40-antibodies detection and sHLA-G quantitation in serum samples. Results: The presence of SV40 antibodies was observed in 6 % of patients affected by MS (N = 4/63), 10 % of OIND (N = 8/77) and 15 % of NIND (N = 9/59), which is suggestive of a lower prevalence in respect to HS (22 %, N = 18/83). MS patients are characterized by higher sHLA-G serum levels (13.9 \ub1 0.9 ng/ml; mean \ub1 St. Error) in comparison with OIND (6.7 \ub1 0.8 ng/ml), NIND (2.9 \ub1 0.4 ng/ml) and HS (2.6 \ub1 0.7 ng/ml) subjects. Interestingly, we observed an inverse correlation between SV40 antibody prevalence and sHLA-G serum levels in MS patients. Conclusion: The data obtained showed a low prevalence of SV40 antibodies in MS patients. These results seems to be due to a generalized status of inability to counteract SV40 infection via antibody production. In particular, we hypothesize that SV40 immune-inhibitory direct effect and the presence of high levels of the immune-inhibitory HLA-G molecules could co-operate in impairing B lymphocyte activation towards SV40 specific peptides

Herpes simplex virus and human cancer. I. Relationship between human cervical tumours and herpes simplex virus type 2

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Detecting recombination in TT virus: a phylogenetic approach

TT virus (TTV) has a remarkable genetic heterogeneity. To study TTV evolution, phylogenetic analyses were performed on 739 DNA sequences mapping in the N22 region of ORF1. Analysis of neighbor-joining consensus trees shows significant differences between DNA and protein phylogeny. Median joining networks phylogenetic clustering indicates that DNA sequence analysis is biased by homoplasy (i.e., genetic variability not originated by descent), indicative of either hypermutation or recombination. Statistical analysis shows that the significant excess of homoplasy is due to frequent recombination among closely related strains. Recombination events imply that the transmission of TTV is not clonal and provide the necessary basis to explain (i) the high degree of genetic divergence between TTV isolates, (ii) the lack of population structure on a world scale, and (iii) the number of highly divergent strains that seems typical of this virus. We show that recombination phenomena can be detected by phylogenetic analyses in very short sequences when a sufficiently large data set is available

Herpes simplex virus latency in immunosuppressed mice

Export | Download | Add to List | More... Microbiologica Volume 7, Issue 3, July 1984, Pages 219-227 Herpes simplex virus latency in immunosuppressed mice. (Article) Rotula, A., Di Luca, D., Gerna, G., Manservigi, R., Tognon, M., Cassai, E. Abstract To determine the role of antibodies in establishing Herpes simplex virus (HSV) latency, immunosuppressed Swiss mice were experimentally infected in the right hind footpad with a HSV-1 x HSV-2 recombinant (C6D) with low virulence. Immunosuppression was induced by repeated intraperitoneal inoculations of Cyclophosphamide (CY) and the production of antibodies to C6D in immunosuppressed animals was monitored by Enzyme Linked Immunosorbent Assay (ELISA). Within 21 days after inoculation, C6D was able to establish a latent infection of lumbosacral spinal ganglia in both normal and CY-treated immunosuppressed animals. The data presented indicate that detectable production of antibodies is not necessary to induce HSV latency in spinal ganglia

Herpes simplex virus and human cancer. II. Search for relationship between labial tumours and herpes simplex type 1

Cellular proliferation was obtained from 30 out of 58 labial tumours examined. None of the cellular cultures obtained presented Herpes Simplex Virus type 1 (HSV-1) specific antigens. Tumour cell cultures demonstrated the same susceptibility to HSV-1 as cells from normal lip tissue, HEp-2 and BHK cells. The presence of HSV-1 DNA was investigated in 12 labial tumours by the blotting technique. The sensitivity of the technique made it possible to reveal 0.5 viral genome equivalent per cell. Viral DNA was not detected in any of the tumours tested

Effect of ketanserin, an inhibitor of 5-HT2 receptors, on the aldosterone-stimulating action of metoclopramide

To estimate the possible involvement of a peripheral serotonergic pathway in the mechanism of the aldosterone-stimulating effect of metoclopramide (M) the plasma aldosterone (PA), renin activity (PRA) and prolactin (PRL) response to M was studied in 6 normal subjects before and after administration of ketanserin (K), a pure, specific, and selective blocking agent of 5-hydroxytryptamine type 2 (5-HT2) receptors. With K preadministration the M-induced increase of PRL was similar to that observed in control conditions, in accordance with the specific and peripheral antiserotonergic action of the drug. K potentiated the PA and PRA elevation in response to M. These data suggest that the PA response to M is not related to M's agonist activity at the peripheral 5-HT2 receptors level. The results further indicate that K can induce an enhancement of the activity of renin-angiotensin-aldosterone system with an higher PRA and PA response to stimulatory action of M

Herpes simplex virus and human cancer. III: Search for relationship of herpes simplex antibodies and cervical dysplasia and labial neoplasia

We employed the enzyme-linked immunosorbent assay (ELISA), indirect hemagglutination (IHA), and complement fixation (CF) methods to measure antibody titer to herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) in patients affected by labial tumors or cervical dysplasias. No relationship of antibody titer to HSV-1 and labial tumors was detected by any of the three methods. Association between antibody titer to HSV-2 and cervical dysplasias was revealed by IHA (p less than 0.05) and ELISA (p less than 0.001); CF tests were negative. Moreover, we assayed for HSV-specific antigens in cell cultures derived from labial tumors and cervical dysplasias. In cultures from labial tumors, it was not possible to detect HSV-specific antigens. Of the 25 cultures derived from cervical dysplasias, HSV antigens were found in only 3 cultures

Mapping of linear epitopes of human papillomavirus type 16: the L1 and L2 open reading frames

Certain types of human papillomavirus (HPV), notably HPV type 16, are associated with flat or inverted proliferative lesions of the cervix uteri that can progress to malignancy. As a first step towards the serological study of the epidemiology of HPV, we have synthesized the entire amino acid sequences of the 2 major viral capsid proteins of HPV type 16, L1 and L2, as a set of 66 synthetic 20-residue peptides with an overlap of 5 amino acids. The peptides were tested for reactivity with IgA, IgG and IgM antibodies in the sera of 30 patients with HPV-16-carrying cervical neoplasms. Both IgG and IgM antibody responses were detected, but most of the reactivity found was of the IgA class. The most immunoreactive peptides were further analyzed for reactivity with sera from 22 patients with parotid gland tumors and with sera from 38 healthy individuals. The L2-encoded protein contained only one major linear epitope, which was not specific for HPV-16-carrying neoplasms. In contrast, the L1-encoded protein contained several epitopes that were regularly immunoreactive with antibodies present in the sera of patients with HPV-16-carrying cervical neoplasms, but only rarely so in the sera of patients with other tumors or of healthy individuals