Herpes simplex Virus 2 (HSV 2) IgM ELISA

Cerastoderma glaucum, a marine bivalve inhabiting Lake Timsah, is surrounded by different pathogenic organisms. The present study evaluates the antimicrobial activities, cytotoxicity and characterization of C. glaucum extracts. Chloroform, methanol and acidic tissue extracts were prepared from C. glaucum collected during winter and summer seasons. Winter acidic extract exhibits potent antimicrobial activities against 21 bacterial, 2 yeast and 2 viral strains. The inhibition zone of this extract ranges from 10 mm against Klebsiella oxytoca, Pseudomonas stutzeri, Globicatella sulfidifaciens and Bacillus (B. badius, B. amyloliquefaciens and B. pumilus) to 24 mm against Shigella flexneri. Also, the inhibition viral activities of this extract at a concentration of 62.5 mu g ml(-1) against Hepatitis A virus and Herpes simplex virus type 1 (HSV-1) are 62.383% and 57.035%, respectively, with low cytotoxicity of 24.030%. Furthermore, winter acidic extract of C. glaucum has the highest total protein contents (9.8 mg ml(-1)) compared with the other extracts. Moreover, the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) indicates the presence of four clear low molecular weight peptides bands; 8.588, 7.237, 4.423 and 2.692 kDa. Fourier transform-infrared (FT-IR) analysis indicates the presence of 12 functional groups of proteins in winter acidic extract of C. glaucum at appropriate wavelengths.

ND10 nuclear bodies, as part of the intrinsic defenses, impose repression on incoming DNA. Infected cell protein 0 (ICP0), an E3 ubiquitin ligase of herpes simplex virus 1 (HSV-1), can derepress viral genes by degrading ND10 organizers to disrupt ND10. These events are part of the initial tug of war between HSV-1 and host, which determines the ultimate outcome of infection. Previously, we reported that ICPO differentially recognizes promyelocytic leukemia (PML) isoforms. ICP0 depends on a SUMO-interaction motif located at residues 362 to 364 (SIM362-364) to trigger the degradation of PML isoforms II, IV, and VI, while using a bipartite sequence flanking the RING domain to degrade PML I. In this study, we investigated how the SUMO-SIM interaction regulates the degradation of PML II and PML II-associated proteins in ND10. We found that (i) the same regulatory mechanism for PML II degradation was detected in cells permissive or nonpermissive to the ICP0-null virus; (ii) the loss of a single SIM362-364 motif was restored by the presence of four consecutive SIMs from RNF4, but was not rescued by only two of the RNF4 SIMs; (iii) the loss of three C-terminal SIMs of ICP0 was fully restored by four RNF4 SIMs and also partially rescued by two RNF4 SIMs; and (iv) a PML II mutant lacking both lysine SUMOylation and SIM was not recognized by ICP0 for degradation, but was localized to ND10 and mitigated the degradation of other ND10 components, leading to delayed viral production. Taken together, SUMO regulates ICP0 substrate recognition via multiple fine-tuned mechanisms in HSV-1 infection. IMPORTANCE HSV-1 ICP0 is a multifunctional immediate early protein key to effective replication in the HSV-1 lytic cycle and reactivation in the latent cycle. ICP0 transactivates gene expression by orchestrating an overall mitigation in host intrinsic/innate restrictions. How ICP0 coordinates its multiple active domains and its diverse protein-protein interactions is a key question in understanding the HSV-1 life cycle and pathogenesis. The present study focuses on delineating the regulatory effects of the SUMO-SIM interaction on ICP0 E3 ubiquitin ligase activity regarding PML II degradation. For the first time, we discovered the importance of multivalency in the PML II-ICP0 interaction network and report the involvement of different regulatory mechanisms in PML II recognition by ICP0 in HSV-1 infection.