Status of NAT screening for HCV, HIV and HBV: experience in Japan.

The first nationwide nucleic acid amplification testing (NAT) for hepatitis B virus (HBV), hepatitis C virus (HCV), and human immunodeficiency virus type 1 (HIV-1) of voluntarily donated blood after serological pre-screening and before release of cellular components and plasma for fractionation was implemented by the Japanese Red Cross Blood Transfusion Services. The NAT screening assay using multiplex reagent is time-saving, cost effective, and labour-saving procedure for all blood and blood products including short-shelf life platelets. During the 50-mini-pool NAT screening of serologically negative donations (February 1, 2001-April 30, 2001), we were able to screen out 112 HBV-positive, 25 HCV-positive, and 4 HIV-1 positive units from blood and blood components.

Intra-parenchymal tumor biopsy using neuroendoscopy with navigation.

BACKGROUND AND OBJECTIVES: Neuroendoscopy has allowed us to biopsy tumors located in the ventricle and the para-ventricle. With this technique we can obtain specimens under direct visual monitoring and examine tumor dissemination. For intra-parenchymal tumors, however, we normally use stereotactic procedures to collect tissues or perform open biopsies. We now report that we have successfully combined neuroendoscopy with navigational guidance to biopsy intra-parenchymal tumors. We explain our methods and discuss the advantages and disadvantages of this technique. CASES AND METHODS: We carried out intra-parenchymal tumor biopsies or resection using neuroendoscopy and guided navigation in three patients. We advanced a transparent sheath, containing a removable inner tube on which a navigation tool was mounted, to target tissues. We directly monitored procedures with a neuroendoscope placed in the transparent sheath. We collected specimens just in front of and in the tumor by forceps. In one patient with bleeding, we used the aspirator tip as a monopolar coagulator. We were also able to check for bleeding along the sheath tract using endoscopy. Without causing any new neurological deficits, we collected tissues for histological diagnosis of astrocytoma in two patients and radiation necrosis in one. CONCLUSIONS: We believe that combining neuroendoscopy with navigation guidance is a safe and precise method for obtaining biopsies of intra-parenchymal tumors. Tumors with rich vasculature will not benefit from this procedure until better hemocoagulation instruments have been developed.

Expression of lymphotoxin gene inserted into Theiler's murine encephalomyelitis virus.

Theiler's murine encephalomyelitis virus (TMEV) belongs to the Picornaviridae genus. DA subgroup strains of this virus induce early, non-fatal polioencephalomyelitis followed by demyelination in the spinal cord, with virus persistence. We investigated the use of DA strain as a vector for the introduction of a foreign gene into the central nervous system. Human lymphotoxin (LT) gene was inserted in the L region, the most upstream part of the polyprotein coding region of DA genome. Expression of LT was demonstrated by an immunoblot and an enzyme-linked immunosorbent assay on BHK-21 cells that were infected with the recombinant virus. In addition, the expressed LT showed cytotoxicity against L-929 cells.

Secondary-phase-modulation method for open-loop fiber-optic gyroscopes.

A proposed method of secondary phase modulation for open-loop fiber-optic gyroscopes is examined in general terms. To detect the rotation rate of a system through a beat-frequency channel, we employ linearly combined signals with different frequencies for the optical phase modulation. We find that the proper combinations of the modulation frequencies can optimize the sensitivity of gyroscopes. With this method we can employ a high-frequency band for optical phase modulations while keeping relative a lower-frequency band of the detection channel. The theoretically derived result is experimentally confirmed by using a lithium-niobate (LiNbO(3)) optical phase modulator. We also discuss the combination setup with an optical integrated-circuit device and digital signal processing.

Full-length sequence of a hepatitis C virus genome having poor homology to reported isolates: comparative study of four distinct genotypes.

Variable genomic sequences have been reported for RNA cloned from hepatitis C virus (HCV)-infected humans and chimpanzees. We found that four distinct genotypes of HCV could be differentially identified by PCR using type-specific primers. Full-length sequences have so far been reported for three of the four HCV genotypes, and we report herewith the sequence of the fourth type obtained from a Japanese blood donor. The entire nucleotide sequence of the HCV isolate (HC-J8) comprised 9481 bases plus a 3'-terminal poly(U) stretch of variable length. Like all previous isolates, the RNA contained a single, long open reading frame for a polyprotein of 3033 amino acids. HC-J8 differed from previously reported HCV isolates by 23.1-33.1% in nucleotide sequence and 15.9-28.8% in amino acid sequence. Based on genomic sequence homologies, a proposed phylogenetic tree of HCV, with a fourth branch represented by HC-J8, allowed a classification of all HCV isolates whose complete or partial sequences are now known. This classification suggests that all or most HCV genome sequences will fall into one of the proposed four types. The classification may be helpful in designing vaccine studies and for serological investigations of possible group- and type-specific antibodies.

The first large-scale nucleic acid amplification testing (NAT) of donated blood using multiplex reagent for simultaneous detection of HBV, HCV, and HIV-1 and significance of NAT for HBV.

The first nationwide nucleic acid amplification testing (NAT) for hepatitis B virus (HBV), hepatitis C virus (HCV), and human immunodeficiency virus type 1 (HIV-1) of voluntarily donated blood after serological pre-screening and before release of cellular components and plasma for fractionation was implemented by the Japanese Red Cross Blood Transfusion Services. From February 1, 2000 to April 30, 2001, specimens from 6,805,010 units of serologically negative donation were screened in minipools of 50 samples within 24 hr after blood donation by NAT using multiplex HBV/HCV/HIV-1 reagent for blood transfusion including short shelf-life platelets. Among them, 112 HBV DNA-positives, 25 HCV RNA positives and 4 HIV-1 RNA positives were screened out and we could prevent transfusion of these NAT positive units. Subtypes/genotypes of HBV DNA, adr/C, adw/A, adw/B, adw/C, ayr/C and ayw/D were found and adr/C was predominant. A total of 61.6 % of them (69/112) were negative by overnight EIA. Sixth three of HBV NAT-positive samples carried virus loads less than 10(4) copies/mL and 92.1 % of them (58/63) were negative by overnight EIA. The virus growth curves of HBV in 6 cases obtained by retrospective and prospective follow-up study showed exponential straight lines in the early stage of serological window periods and the log times of HBV growth (10 fold increase) in serological window period were between 4.6 and 7.6 days. NAT screening with highly sensitive reagents in pool of specimens is useful to exclude blood units with low level of HBV and HBV mutants from blood transfusion.