Direct sequencing and characterization of a clinical isolate of Epstein-Barr virus from nasopharyngeal carcinoma tissue by using next-generation sequencing technology.

Epstein-Barr virus (EBV)-encoded molecules have been detected in the tumor tissues of several cancers, including nasopharyngeal carcinoma (NPC), suggesting that EBV plays an important role in tumorigenesis. However, the nature of EBV with respect to genome width in vivo and whether EBV undergoes clonal expansion in the tumor tissues are still poorly understood. In this study, next-generation sequencing (NGS) was used to sequence DNA extracted directly from the tumor tissue of a patient with NPC. Apart from the human sequences, a clinically isolated EBV genome 164.7 kb in size was successfully assembled and named GD2 (GenBank accession number HQ020558). Sequence and phylogenetic analyses showed that GD2 was closely related to GD1, a previously assembled variant derived from a patient with NPC. GD2 contains the most prevalent EBV variants reported in Cantonese patients with NPC, suggesting that it might be the prevalent strain in this population. Furthermore, GD2 could be grouped into a single subtype according to common classification criteria and contains only 6 heterozygous point mutations, suggesting the monoclonal expansion of GD2 in NPC. This study represents the first genome-wide analysis of a clinical isolate of EBV directly extracted from NPC tissue. Our study reveals that NGS allows the characterization of genome-wide variations of EBV in clinical tumors and provides evidence of monoclonal expansion of EBV in vivo. The pipeline could also be applied to the study of other pathogen-related malignancies. With additional NGS studies of NPC, it might be possible to uncover the potential causative EBV variant involved in NPC.

Isolation and Characterization of a Neurotrophic Factor-like Substance from Venom of Agkistrodon acutus.

A neurotrophic factor-like substance from Agkistrodon acutus was isolated by ion exchange and gel filtration chromatography and was found to be homogenous by electrophoresis. Itsmolecular weight was estimated to be approximately 26 kD by gel filtration. A characteristic of the substance was that the protein consisted of two subunits which were bound one to another noncovalently. The analytical isoelectric focusing revealed its isoelectric point of 7.8. The biological activity of the substance was comparable to that of mouse 2.5 S NGF. It rescued PC12 cells from apoptosis at the concentration interval of 1 100 &mgr;g/L and could induce PC12 cells differentiate to sympathetic-like neurons.