Molecular Analysis of Fiji Disease Fijivirus Genome Segments 1 and 3

Fiji disease fijivirus (FDV) genomic segments 1 (S1) and 3 (S3) were completely sequenced. FDV S1 comprised 4532nt and was predicted to encode a 170.6kDa protein. FDV S3 comprised 3623nt and was predicted to encode a 135.5kDa protein. The terminal sequences of S1 and S3 were 5 AAGUUUUU......CAGCUAGCGUC 3 and 5 AAGUUUUU......CAGCAGAUGUC 3, respectively, and located immediately adjacent to these sequences were 12bp imperfect inverted repeats. The predicted translation product of FDV S1 showed highest similarity to Rice black-streaked dwarf virus (RBSDV) S1 and is thought to encode the viral RNA-dependent RNA polymerase (RdRp). The predicted translation product of FDV S3 was found to be most similar to RBSDV S4 which is thought to encode the 'B-spike' protein. The FDV sequence contained an ATP/GTP binding motif and a leucine zipper motif, but these motifs were not found in the RBSDV sequence. Phylogenetic analysis based on the amino acid sequences of the RdRp of FDV S1 and other reoviruses revealed that the fijiviruses form a cluster close to the oryzaviruses. The RdRp sequences were grouped into genera that were consistent with the current reovirus classification scheme that is based on physico-chemical and biological properties.     

The complete nucleotide sequence of isolate BYMV-GDD of <Emphasis Type="Italic">Bean yellow mosaic virus</Emphasis>, and comparison to other potyviruses

Summary. The complete nucleotide sequence of Bean yellow mosaic virus (BYMV) gladiolus isolate GDD was determined and compared to broad bean isolates BYMV-MB4 and BYMV-S. The BYMV-GDD genome (9528nt) was more similar to BYMV-MB4 (9532nt) than to BYMV-S (9547nt), which has atypical symptom expression and host range. The greatest variability occurred in the 5 untranslated region, P1 protein, and NIa-VPg protein, the N-terminal two thirds of HC-Pro, and the C-terminal one third of P3. Each of these regions has been correlated with symptom or host differences between isolates of other potyviruses, and may contribute to the atypical nature of BYMV-S.     

Mother-Baby psychiatric units (MBUs): National data collection in France and in Belgium (1999–2000)

Summary The French version of the Marcé checklist was used to collect data for 176 joint admissions to 11 psychiatric mother-baby units in 1999 and 2000. Mean age of the babies at admission ranged from 4 to 16 weeks. Two units also admitted older children. Mothers admitted were diagnosed with schizophrenia or chronic delusional disorders (n=44), acute transitory psychosis Bouffée délirante (n=20), bipolar disorders (n=20), depressive illness (n=38), personality disorders or intellectual disability (n=39), and other disorders (n=15). The mean duration of hospitalisation was 11 weeks. Units that also offered day-care admission in the same or a near-by unit had shorter mean admissions. More than half the womens partners (or babies fathers) had mental health problems. Women with schizophrenia or chronic delusional disorders and personality disorders or intellectual disability remained hospitalised longer, improved less, and were more often separated from their babies, or discharged with supervision, than women admitted with other diagnoses.     

Hemodilution with liposome-encapsulated low-oxygen-affinity hemoglobin facilitates rapid recovery from ischemic acidosis after cerebral ischemia in rats

Liposome-encapsulated hemoglobin (LipoHb) with low oxygen affinity (P₅₀ = 40–50mmHg) has been developed. The purpose of this study was to evaluate the effects of the LipoHb on incomplete cerebral ischemia. Wistar rats were randomly assigned to one of the following three groups: (A) exchange transfusion with LipoHb solution (Hb = 6g/dl) (LipoHb, n = 7), (B) exchange transfusion with rat red blood cell (RBC) solution (Hb = 6g/dl) (RBC, n = 7), (C) no exchange transfusion (control, n = 7). Forebrain ischemia was induced for 9min by bilateral carotid artery occlusion combined with a decrease in the mean arterial pressure (MAP) to 40mmHg. ³¹P-magnetic resonance spectroscopy was performed during ischemia and 60min of reperfusion. After exchange transfusion, the MAP increased in the LipoHb group and decreased in the RBC group (LipoHb versus RBC; P = 0.0028). During ischemia, intracellular pH (pHi) rapidly decreased in all groups; after reperfusion, the pHi recovery to preischemic levels was more rapid in the LipoHb group than in the RBC group (P < 0.05). Phosphocreatine and -adenosine triphosphate decreased during ischemia and returned to the preischemic level in all groups following reperfusion. Inorganic phosphate (Pi) increased during ischemia and decreased to the normal value after reperfusion. The LipoHb group had a smaller production of Pi than the other two groups and demonstrated a rapid normalization, although the differences were not significant. Hemodilution with liposome-encapsulated low-oxygen-affinity hemoglobin facilitates rapid pHi recovery from incomplete forebrain ischemia in the rat.     

The relationship between HPV16 and expression of CD44v6, nm23H1 in esophageal squamous cell carcinoma

Summary. The esophageal squamous cell carcinoma (ESCC) has high incidence in Shaanxi Province of China. More and more researches indicated that human papillomavirus type 16 (HPV16) might play an important role in carcinogenesis of ESCC but the relationship between HPV16 and CD44v6, nm23H1 has not been elucidated. HPV16 was detected by amplifying HPV16 E6 gene through polymerase chain reaction (PCR) method and the expression of CD44v6, nm23H1 in 40 ESCCs and fifteen normal esophageal mucosa (NEM) from Shaanxi Province was examined by Streptavidin-Peroxidase (SP) method using monoclonal antibody specific to CD44v6 and nm23H1. The positive rates of HPV16 E6 gene, CD44v6 and nm23H1 were 60% (24/40), 65% (26/40) and 45% (18/40) respectively in ESCCs and 26.67% (4/15), 33.33% (5/15) and 86.67% (13/15) respectively in NEMs. There exited statistical difference for HPV16, CD44v6 and nm23H1 between NEMs and ESCCs respectively (p<0.05). The relationship between HPV16 and the expression of CD44v6 in ESCCs was statistical significance (P=0.021), but no significant correlation was found between HPV16 and the expression of nm23H1 (P=0.436) in ESCCs. The infection rate of HPV16 had no statistical difference in all pathological features we observed, but the expression rates of CD44v6 and nm23H1 had statistical correlation with invasion (p=0.001, 0.013) and lymph nodes metastasis (p=0.014, 0.002) respectively. In different histology grade of ESSCs, the relationship between HPV16 and CD44v6 was statistical significance in grade I (p=0.044) but was not in grade II (p=0.165) and grade III (p=0.658), however as to the expression of nm23H1 there exited no statistical significance in all histology grades of ESCC (p>0.05). The expression rates of CD44v6 and nm23H1 were statistically different between grade I and II (p=0.004, 0.016) respectively and between grade I and grade III (p=0.014, 0.020), but not statistically different between grade II and III (p=0.792, 0.943) respectively. Our data firstly suggested that there existed the statistical relationship between the infection of HPV16 and the expression of CD44v6 in ESCCs and that HPV16 may be involved in invasion and metastasis of ESCC.     

Pattern of <Emphasis Type="Italic">N</Emphasis> gene-mediated systemic hypersensitive response and turnover of viral replicase protein in tobacco

Summary. We examined the pattern of the N gene-mediated systemic hypersensitive response (HR), which was induced by tobacco mosaic virus upon temperature shift, and analyzed the distribution of the coat protein and the virus-encoded 126kDa replicase protein (126K protein) by immunoblot analysis. In the middle- and lower-positioned leaves, HR occurred in the advancing edge of the infected area, where we detected both the coat protein and the 126K protein. In the areas between the main vein and the advancing edge of these leaves, we observed no HR and did not detect 126K protein, though virus was present in these areas. In the upper-positioned mosaic leaves, patterns of the HR were different depending on the leaf age. In these mosaic leaves, the mechanism preventing the virus from invading dark green tissue seemed to be broken down in 8–14 days old leaves, and HR was observed only in the tissue just invaded by the virus, where we detected the 126K protein. Those results suggested that the viral 126K protein was present when the viral replication was taking place, and easily degraded when the amount of the virus was saturated in the cells.     

The complete nucleotide sequence of the genome RNA of Lily symptomless virus and its comparison with that of other carlaviruses

Summary. The complete genomic nucleotide sequence and genome structure of Lily symptomless virus (LSV), a lily-infecting carlavirus, have been obtained. The genome of the Korean strain of LSV, LSV-Kr, was 8,394 nucleotides long and contained six open reading frames (ORFs) coding for proteins of Mr 220kDa (1,948aa), 25kDa (228aa), 12kDa (106aa), 7kDa (64aa), 32kDa (291aa) and 16kDa (140aa) from the 5 to 3 end, respectively, which is typical of carlaviruses. Genetic heterogeneity was observed in the ORF1 gene. A total of 221 of 5,847 nucleotides (nt) were heterologous in the ORF1 of replicase; 162nt portions were silent and 59nt resulted in amino acid changes. This heterogeneity indicates that the LSV-infecting lily plants contained a genetically heterogeneous population of LSV (quasispecies). Overall similarities to those of other carlaviruses for the six ORFs of LSV were from 76.1% to 31.6% and from 87.3% to 13.7%, at nucleotide and amino acid levels, respectively. The ORF1 replicase gene of LSV shares 40.9% to 56.8% and 48.9% and 58.6% identities with that of 5 other carlaviruses at the amino acid and nucleotide levels, respectively. LSV was closest to Blueberry scorch virus (BlScV) in this ORF, among the carlaviruses for which sequence information is available. The three triple gene blocks (ORF2-4), ORF5 (coat protein) and 3-proximal 16kDa ORF6 genes were further analyzed, and phylogenetic trees for the coding regions indicate that the LSV was the most closely related to Kalanchoe latent virus and BlScV. This is the first report of the complete nucleotide sequence and genome structure of LSV.Received December 13, 2002; accepted May 14, 2003 Published online July 17, 2003     

Sequence analysis of the fragment of the phosphoprotein gene of Polish distemper virus isolates

Summary. The nucleotide sequence analysis of the 429bp fragment of the P gene of 11 Polish field isolates of Canine distemper virus (CDV), reference strains and other virus isolates available in the GenBank was the aim of the studies. High homology between all dog strains from east-southern region of Poland and reference strains of CDV was demonstrated. It was estimated as 97–100% for CDV-OND; 96.7–99.8% for CDV-Rock; 96.7–99.8% for CDV-LED and 96.3–97.9% for A75-17. The 100% homology of the nucleotide sequence was observed between CDV Pulawy 92, CDV Pulawy 97 and the reference CDV-OND. The homology between CDV-OND and viruses isolated from the mink and ferret was estimated as 97.7% and 98.4%, respectively. Virus strains isolated from blue foxes demonstrated the highest homology to CDV-OND – equal to 97.7% for DV 79 and 99.5% for DV 92. The fox isolate from 1992 had higher level of homology to dog isolates (96.5–99.5%) than the strain isolated from the fox in 1979 (97.2–98.8%). The phylogenetic tree has two main lineages representing two separated genetic groups: I containing PDV and II containing all distemper virus strains isolated from terrestrial carnivores. CDV strains isolated from dogs from Pulawy region between 1992–1998 and from the fox (DV 92) formed the separate lineage containing also reference strains. They differed from the native isolates from the mink and ferret as well as from Japanese strains of CDV.Received October 29, 2002; accepted April 2, 2003 Published online June 11, 2003     

Analysis of cytosolic and lysosomal pH in apoptotic cells by flow cytometry

Several reports indicate that the cytosol is acidified during apoptosis although the mechanism is not yet fully elucidated. The most acidic organelle found in the cell is the lysosome, raising the possibility that lysosomal proton release may contribute to the cytosolic acidification. We here describe methods for measurement of the cytosolic and lysosomal pH in U937 cells by a dual-emission ratiometric technique suitable for flow cytometry. Cytosolic pH was analysed in cells loaded with the fluorescent probe BCECF, while lysosomal pH was determined after endocytosis of FITC-dextran. Standard curves were obtained by incubating cells in buffers with different pH in the presence of the proton ionophore nigericin. Apoptosis was induced by exposure of cells to 10ng/ml TNF- for 4h, and apoptotic cells were identified using a fluorescent marker for active caspases. By gating of control and apoptotic cells, the cytosolic and lysosomal pH were calculated in each population. The cytosolic pH was found to decrease from 7.2 ± 0.1 to 5.8s±0.1 and the lysosomal increased from 4.3±0.4 to 5.2±0.3. These methods will be useful in future attempts to evaluate the involvement of lysosomes in the acidification of the cytosol during apoptosis.     

Changes in level of virus accumulation and incidence of infection are critical in the characterization of <Emphasis Type="Italic">Rice tungro bacilliform virus</Emphasis> (RTBV) resistance in rice

Summary. Analysis by enzyme-linked immunosorbent assay showed that Rice tungro bacilliform virus (RTBV) accumulated in a cyclic pattern from early to late stages of infection in tungro-susceptible variety, Taichung Native 1 (TN1), and resistant variety, Balimau Putih, singly infected with RTBV or co-infected with RTBV+Rice tungro spherical virus (RTSV). These changes in virus accumulation resulted in differences in RTBV levels and incidence of infection. The virus levels were expressed relative to those of the susceptible variety and the incidence of infection was assessed at different weeks after inoculation. At a particular time point, RTBV levels in TN1 or Balimau Putih singly infected with RTBV were not significantly different from the virus level in plants co-infected with RTBV+RTSV. The relative RTBV levels in Balimau Putih either singly infected with RTBV or co-infected with RTBV+RTSV were significantly lower than those in TN1. The incidence of RTBV infection varied at different times in Balimau Putih but not in TN1, and to determine the actual infection, the number of plants that became infected at least once anytime during the 4wk observation period was considered. Considering the changes in RTBV accumulation, new parameters for analyzing RTBV resistance were established. Based on these parameters, Balimau Putih was characterized having resistance to virus accumulation although the actual incidence of infection was >75%.Received December 18, 2002; accepted March 19, 2003 Published online June 5, 2003     

Development of an artificial synapse using an electrochemical micropump

Improving the resolution of artificial sensory organs requires an interface that receives external information from electronic circuits and stimulates appropriate neurons individually in response to that information. The method of electric stimulation in available artificial sensory organs is fairly nonselective; therefore, we developed a method of chemical stimulation of neurons using a neurotransmitter containing an electrochemical micropump powered by the bubbling that occurs during water electrolysis. The micropump contains a glass nozzle with a tip 10µm in diameter. Two blackened platinum electrodes for the electrolysis were inserted into the body of the pump, which was filled with neurotransmitter solution. The distance between a neuron of the gastropod Aplysia and the tip of the nozzle was adjusted to about 100µm. A potential difference of 3.0V was applied to the electrodes to propel the solution toward the neuron while its membrane potential was monitored. Administration of 1-mM acetylcholine to a resting neuron caused neural firing only when the voltage was applied for 0.5s and without a time lag. During administration of 50-mM -aminobutyric acid to spontaneously firing neurons, the firing disappeared with a time lag of 1s after application of 3.0V. We concluded that an electrochemical micropump can be applied for rapid neurotransmitter administration to control the excitation and inhibition of neurons. This simple pump can be miniaturized to create synapses in artificial sensory organs.     

A minor outer capsid protein,P9, of <Emphasis Type="Italic">Rice dwarf virus</Emphasis>

Summary Partial amino acid sequence of a minor 30kDa polypeptide in purified Rice dwarf virus (RDV) was identical to the deduced amino acid sequence encoded by the dsRNA segment S9 of the virus. This polypeptide was specifically detected by Western blotting analysis with antibodies raised against the product of S9 expressed in a baculovirus system. Treatment of purified RDV particles with a relatively higher concentration of MgCl₂ removed the polypeptide from core particles together with other outer capsid proteins. These results demonstrate that the 30kDa polypeptide is a minor outer capsid protein that is encoded by genome segment S9 of RDV. This protein was named P9 protein.Received April 23, 2003; accepted May 23, 2003 Published online August 7, 2003     

Relapse of depression during pregnancy following antidepressant discontinuation: a preliminary prospective study

Summary. Objective: Pregnancy has frequently been referred to as a time of emotional well-being for patients. However, systematic data about the risk for relapse of depression during pregnancy are sparse.Method: We completed a longitudinal cohort study of thirty-two (N=32) women with histories of depression who were euthymic at conception and who either discontinued or attempted to discontinue antidepressant therapy proximate to conception. Subjects were prospectively followed across pregnancy once per trimester using structured clinical interviews. Rates of relapse and time to relapse were examined. Factors distinguishing the population with respect to risk for relapse including demographic characteristics and illness history were also examined.Results: Seventy-five percent (N=24) of patients relapsed during pregnancy. The majority of relapses (79%, N=19) occurred in the first trimester, and relapse was more prevalent in women with histories of more chronic depression.Conclusions: Pregnancy is not protective with respect to risk for relapse of depression. Careful treatment planning is necessary for those women on antidepressants who plan to conceive or who become pregnant.     

Nano-scaled hydroxyapatite/polymer composite IV. Fabrication and cell adhesion properties of a three-dimensional scaffold made of composite material with a silk fibroin substrate to develop a percutaneous device

Nano-scaled sintered hydroxyapatite (HAp) particles with an a-axis length of 87 ± 23nm, a c-axis length of 236 ± 81nm, and a mean aspect ratio (c/a) of 2.72 were covalently linked onto a silk fibroin (SF) substrate chemically modified by graft polymerization with -methacryloxypropyl trimethoxysilane (MPTS). Graft polymerization with poly(MPTS) on SF was conducted by free-radical initiation in a water solvent with pentaethylene glycol dodecyl ether as a nonionic surfactant. The alkoxysilyl groups of the graft polymers avoided hydrolysis and maintained their activity in coupling with the hydroxyl groups on the HAp surface despite the use of water as the reaction solvent. The weight gain of poly(MPTS) on SF increased with increasing the reaction time, eventually reaching a plateau value of about 15wt% after 50min of reaction time. After HAp covalent coating, the particles separated or aggregated into several crystals, as shown by scanning electron microscopic observation. L929 fibroblast cells adhered more plentifully on HAp-coated SF compared to untreated SF and hydrolyzed poly(MPTS)-grafted SF during 24h or 48h of incubation. The cells adhered only on the HAp surface but not at all on the dehydrated grafted surface of SF without HAp. A button-shaped prototype for a percutaneous device was manufactured by transplantation of HAp-coated SF fibers of about 100µm in length onto silicone moldings using an adhesive, and the device showed good cell adhesiveness.     

Actin filaments around endothelial fenestrae in rat hepatic sinusoidal endothelial cells

The presence of microfilaments in the vicinity of sinusoidal endothelial fenestrae (SEF) suggests that the cytoskeleton of liver sinusoidal endothelial cells (LSEC) plays an important role in the modulation of SEF. In this study, we investigated actin filaments around SEF in LSECs. Monolayers of LSEC culture were established by infusing a rat liver with collagenase for 30min and then culturing in RMPI medium for 24h. Cells were reacted with 0.1% Triton X for 5s and 15% glycerinated PHEM buffer (60mM PIPES, 25mM HEPES, 10mM EGTA, 2mM MgCl, pH 6.9) containing heavy meromyosin for 10min and observed under a transmission electron microscope. By electron microscopy with the modified heavy meromyosin decorated reaction, actin filaments were clearly demonstrated around SEF in LSEC.     

Usefulness of low-priming-volume cardiopulmonary bypass circuits and dilutional ultrafiltration in neonatal open-heart surgery

In neonate open-heart surgery, cardiopulmonary bypass (CPB) with extreme hemodilution induces an increased capillary permeability and accumulation of extravascular fluid, resulting in organ dysfunction. We evaluated the effects of a reduced priming volume for CPB and dilutional ultrafiltration (DUF) during neonatal open-heart surgery. Nineteen consecutive neonates with complete transposition of the great arteries who underwent an arterial switch operation were retrospectively assigned into two groups: the high-priming-volume circuit group (group A, n = 9) and the low-priming-volume circuit group (group B, n = 10). Patients in group B underwent surgery with a miniaturized CPB circuit and using the DUF technique. The priming volume of group B was nearly two-thirds that of group A. The water balance value after CPB and surgery was significantly lower in group B (–126 ± 118ml, –116 ± 116ml) than in group A (88 ± 218ml, 83 ± 165ml). Systolic blood pressure just after CPB was higher in group B (67.9 ± 9.1mmHg) than in group A (55.4 ± 10.3mmHg). Postoperative ventilatory support was shorter in group B (45 ± 19h) than in group A (68 ± 27h). In neonatal cardiac surgery, low-priming-volume CPB circuits and DUF improve the water balance during surgery and may attenuate any inflammatory reaction, which would help preserve postoperative organ function.     

Detection of enteric viruses and bacterial indicators in German environmental waters

Summary. A German mining lake and the supplying surface waters, which are located downstream of a sewage plant, were examined regarding their microbiological and virological quality. Between October 2002 and September 2003, specific PCR methods were used to determine the occurrence of enteric viruses in 123 water specimens drawn at different sites downstream of the waste water treatment plant and in 9 samples from the sewage plant influent. Detection rates in sewage plant effluents and surface water samples depended on sampling sites and were: 29–76% for enterovirus (EntV), 24–42% (astrovirus, AstV), 15–53% (norovirus, NV), 3–24% (rotavirus, RoV), 5–20% (hepatitis A virus, HAV) and 20% (adenovirus, AdV). AstV genome load of selected samples was between 3.7×10³ to 1.2×10⁸ genome equivalents per liter (gen.equ./l), depending on sampling location; NV average genome load ranged from 1.8×10⁴ to 9.7×10⁵gen.equ./l. Cell culture methods showed that three out of 18 PCR positive samples contained infectious EntV. Even though microbiolical parameters such as Escherichia coli, enterococci and coliphages indicated acceptable microbiological water quality, the virological data of this study suggest the possibility that surface waters may be a source for enteric viral infections.     

Development of a dialyzer with enhanced internal filtration to increase the clearance of low molecular weight proteins

Accumulated low molecular weight proteins in hemodialysis patients require a high-flux dialyzer. There have been several methods proposed for enhancing internal filtration, including narrowing the inside diameter of the hollow fibers, lengthening the fibers, and increasing the fiber density ratio. We tried to enhance the internal filtration by increasing the pressure drop in the dialysate compartment through increasing the fiber density ratio. If the fiber density ratio is too high, however, an irregular dialysate path may result, thus decreasing dialysis performance. Therefore, we took note of the shape of the inner housing and added a short taper structure, which improved the dialysate path dramatically. Consequently, we developed an internal filtration-enhanced dialyzer (APS-Prototype) to improve dialysis performance. The internal filtration rate in water (measured by Doppler ultrasound) was 13.2l/session for the APS-Prototype and 5.3l/session for the APS-15E. The amount of 1-microglobulin (1-MG) in bovine plasma was 0.34g for the APS-Prototype and 0.11g for the APS-15E. In addition, the amount of 1-MG in vivo was 29.0% ± 5.8% for the APS-Prototype, significantly higher than that for the APS-15E (13.6% ± 1.9%). The desirable loss of albumin is 2–4g in hemodiafiltration, and it was 3.92 ± 1.03g for the APS-Prototype. The prototype showed excellent solute removal performance with no clinical or engineering problems.     

Determination of complete nucleotide sequence of <Emphasis Type="Italic">Hibiscus latent Singapore virus</Emphasis>: Evidence for the presence of an internal poly(A) tract

Summary. We have sequenced the complete genome of a hibiscus-infecting tobamovirus, Hibiscus latent Singapore virus (HLSV). The experimental host range of HLSV is similar to that of another distinct species of hibiscus infecting tobamovirus, Hibiscus latent Fort Pierce virus (HLFPV). The genomic structure of HLSV is similar to other tobamoviruses in general. It consists of a 5 untranslated region (UTR), followed by ORFs encoding for a 128kDa protein and a 186kDa readthrough protein, a 30kDa movement protein (MP), 18kDa coat protein (CP) and a 3 UTR. The unique feature of HLSV is the presence of a poly(A) tract within its 3 UTR. In our previous work, we have reported MP and CP sequences of HLSV and its phylogenetic analysis. Here we report the complete nucleotide sequence of HLSV, phylogenetic analysis of the nucleotide and amino acid sequences of 128/186kDa ORFs and the presence of a uniquely located poly(A) tract within the 3 UTR.The GenBank accession numbers of the sequences reported in this paper are AF400156, AF400157 and AY497578, respectively.     

Sequence Analysis of the Complete Genome of Rice Black-Streaked Dwarf Virus Isolated from Maize with Rough Dwarf Disease

The complete nucleotide sequences of 10 genomic segments (S1–S10) from an isolate of rice black-streaked dwarf virus causing rough dwarf disease on maize (RBSDV-Hbm) in China were determined, a total of 29,142 base pairs (bp). Each segment possessed the genus-specific termini with conserved nucleotide sequences of (+) 5-AAGUUUUU......CAGCUNNNGUC-3 and a perfect or imperfect inverted repeat of seven to eleven nucleotides immediately adjacent to the terminal conserved sequence. While the coding strand of most RBSDV-Hbm segments contained one open reading frame (ORF), there were two non-overlapping ORFs in S7 and S9, and one small overlapping ORF downstream of the major ORF in S5. Homology comparisons suggest that S1 encodes a RNA-dependent RNA polymerase (RdRp), with 63.5% and 32.6% identity to the putative RdRp encoded by Fiji disease virus (FDV) and Nilaparvata lugens reovirus (NLRV), respectively. The proteins encoded by S2, S3, and S4 showed various degrees of similarity to those encoded by the corresponding segments of FDV or NLRV. In S5 and S6, low identities were found to those of FDV only, but not to NLRV. Sequence analyses showed that RBSDV-Hbm had the most similarities in the genome organizations and the coding assignments with a RBSDV isolated from rice in China, in which each pair of the corresponding segments shared sequence identities of 93.8–98.9% and 93.5–100% at nucleotide or amino acid levels, respectively. In addition, phylogenetic analyses suggested that RBSDV-Hbm had the closest evolutionary relationship to RBSDV in Fijivirus.