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.     

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.     

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     

Strains of coxsackie virus B4 differed in their ability to induce acute pancreatitis and the responses were negatively correlated to glucose tolerance

Summary. The outcome of infections with three CVB4 strains known to replicate in human pancreatic islet cells (E2, V89 4557 and VD2921) and one CVB3 strain (Nancy) in CBA/J mice with regard to viral replication, inflammation and glucose tolerance was studied. Isolation of virus from hearts, livers and pancreata was performed 7, 14 and 21 days post infection (pi). All strains could be equally well isolated from the pancreata and all but one strain, V89 4557, could be isolated from the hearts. The titers of neutralizing antibodies in the mice infected with the CVB4 strain V89 4557 were significantly lower than in mice infected with the other strains (p<0.01). Even though virus was isolated from both heart and pancreata, immunohistochemical staining only revealed inflammatory cells in the latter. Seven days pi there was a significant difference between the strains in this respect i.e. mice infected with CVB3 and the E2 strain revealed more CD4⁺ lymphocytes, macrophages and granulocytes compared to mice infected with the other CBV strains (p<0.05). Glucose tolerance tests performed at day 14 and at day 115 pi revealed normal kinetics of glucose absorption from the blood in the control mice and in mice infected with the strains that induced severe inflammation of the pancreas (E2 and CVB3). In contrast, the glucose clearance in mice infected with the CVB4 strains V89 4557 and VD2921 were significantly impaired compared to uninfected controls and compared to mice infected with the other CVB strains (p<0.01). Mice infected with CVB4 strains V89 4557 also had a significantly impaired clearance of glucose 120min after injection (p<0.05) even though no virus could be isolated and no inflammation was detected in the pancreata at that time point. These results show that there is a clear strain difference with regard to the ability to affect clearance of glucose from the blood as late as 115 days pi as well as the degree of inflammation in the pancreas. This indicates that the in vitro diabetogenic strains (VD2921 and V89 4557) also in vivo can induce a pre-diabetic state in CBA/J mice.     

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.     

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 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.     

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.     

RGD-mutants of B-lymphotropic polyomavirus capsids specifically bind to αvβ3 integrin

Summary. Integrins are a family of cell surface proteins that function as receptors for extracellular matrix ligands and for some viruses. A subset of integrins recognises peptide sequences containing arginine-glycine-aspartic acid (RGD) motifs as ligands. The B-lymphotropic polyomavirus (LPV) has a non-enveloped capsid that recognises a sialylated cell surface receptor. To change the receptor binding specificity we have replaced sets of three amino acids in three predicted surface loops of the major capsid protein VP1 of the B-lymphotropic polyomavirus LPV by RGD.Ten mutants gave rise to the expected 40kDa VP1 protein upon expression from a baculovirus vector in insect cells. Five of the VP1 mutants representing all three surface loops have retained the ability to spontaneously assemble to capsids in the nuclei of the insect cells. Structural changes of the mutant capsid surface were shown by differential reactivity with a set of 7 neutralising monoclonal antibodies that recognise conformational surface epitopes of wildtype LPV virions. In addition all mutant capsids had lost specific binding to the LPV receptor.Three mutant capsids of one loop (BC) showed specific binding to v3 integrin but not to integrins v5, v6, or to IIb3 known also to recognise RGD containing peptide sequences. This selective binding of the mutant capsids could be inhibited by synthetic peptides that specifically bind to v3 integrin with IC₅₀ values between 10 and 40nM.     

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.     

Identification and Characterization of the UL24 Gene Product of Herpes Simplex Virus Type 2

The UL24 gene of herpes simplex virus type 2 (HSV-2) is predicted to encode a 281 amino acid protein with a molecular mass of 30.5kDa. In this study, the HSV-2 UL24 gene product has been identified by using a rabbit polyclonal antiserum produced against a recombinant protein containing the full-length UL24 gene product of HSV-2 fused to glutathione-S-transferase. The antiserum reacted specifically with a 32kDa protein in HSV-2 186-infected Vero cells and with 31 and 32kDa proteins in UL24-expressing Cos-7 cells. Accumulation of UL24 protein to detectable levels required viral DNA synthesis, indicating that the protein was regulated as a late gene. UL24 protein was found to be associated with purified HSV-2 virions and C capsids. Indirect immunofluorescence analysis demonstrated that the UL24-specific fluorescence was detected in perinuclear regions of the cytoplasm and/or in the nucleus as small discrete granules from 9h post infection (hpi). Furthermore, the UL24 protein expressed singly was detected predominantly in the nucleus and slightly in the cytoplasm at 24h after transfection, with branch-like cytoplasmic protruding structures. Strong nucleolus staining was visible in partial cells.     

The complete nucleotide sequence of <Emphasis Type="Italic">Plum pox virus</Emphasis> isolates from sweet (PPV-SwC) and sour (PPV-SoC) cherry and their taxonomic relationships within the species

Summary. Plum pox virus (PPV) sweet (SwC) and sour (SoC) cherry isolates were the first PPV isolates to be recovered from natural infection in sweet and sour cherry plants, respectively. Their complete nucleotide sequences have been determined finding a deduced genome organisation typical for PPV species. Both genomes are 9795 nucleotides long, excluding the 3 terminal poly(A) tail, and contain an open reading frame of 9432nt, encoding a polyprotein of 3143 amino acids. The nucleotide and predicted amino acid sequences of PPV-SwC and SoC have been pairwise compared with available sequences of different PPV strains. Although a very high similarity exists between the whole genomes and polyproteins of the two cherry isolates, high levels of divergence have been calculated with sequences of PPV-M, D and EA isolates. In particular, the most considerable divergence has been found in part of 5 non coding region, in regions encoding P1, P3+6K1, 6K2 and NIa-VPg proteins as well as in the N-terminal domain of the coat protein. Phylogenetic analysis have been undertaken in order to establish the taxonomic localisation of SwC and SoC isolates within PPV species, showing that they are always clustered together and separated from the rest of PPV strains, being clearly the most distant.Received February 26, 2003; accepted June 18, 2003 Published online August 18, 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.     

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.     

Carboxy-terminally truncated Dengue 4 virus envelope glycoprotein expressed in <Emphasis Type="Italic">Pichia pastoris</Emphasis> induced neutralizing antibodies and resistance to Dengue 4 virus challenge in mice

Summary. We have expressed a recombinant Dengue 4 virus envelope glycoprotein (E4rec), truncated at its C-terminus by 53 amino acids, in Pichia pastoris. The presence of E4rec was confirmed by Western-blot using anti-DEN 4 hyper immune mouse ascitic fluid. E4rec migrated during SDS-PAGE as a 64kDa protein. Treatment with endoglycosidases showed that the E protein was modified by the addition of short mannose chains and the absence of hyperglycosylation. When administered to BALB-C mice, E4rec elicited a DEN 4 neutralizing antibody response haemagglutination inhibition antibodies and specific memory T cell response. Mice immunized were also significantly protected against lethal DEN 4 virus challenge (86.6%, p<0.001).Received November 8, 2003; accepted June 7, 2003 Published online August 7, 2003     

Nonocclusive mesenteric ischemia after cardiopulmonary bypass

Nonocclusive mesenteric ischemia (NOMI) is a rare abdominal pathology caused by mucosal hypoperfusion without actual obstruction to the mesenteric arteries. We present a case of NOMI after a cardiopulmonary bypass operation. The patient was a 79-year-old woman with a history of hypertension and diabetes mellitus. A coronary bypass operation was performed with stable hemodynamic conditions, and continuous venovenous hemodialysis was performed on the second postoperative day because of renal insufficiency. After 24h of hemodialysis, the hematocrit level increased from 29.1% to 36.1%. The patient had some vague abdominal pain on the third postoperative day with abnormal laboratory values: leukocytes 17.10 × 10³/µl, creatine kinase 1085U/l, glutamic-oxyloacetic transaminase 6188U/l, and lactate dehydrogenase 8695U/l. Selective angiography showed diffuse stenosis of the superior mesenteric artery (SMA) without any occlusive findings on the major branches; the patient was therefore diagnosed with NOMI. An infusion of urokinase and prostaglandin E1 was started; however, disseminated intravascular coagulopathy had developed and the patient died on the 21st postoperative day as a result of multiple organ failure. The autopsy demonstrated extensive necrosis and hemorrhage in the small intestine without any occlusive findings on the major branches of the SMA.     

Evidence for recombination among isolates of <Emphasis Type="Italic">Tobacco leaf curl Japan virus</Emphasis> and <Emphasis Type="Italic">Honeysuckle yellow vein mosaic virus</Emphasis>

Summary Complete nucleotide sequences of Tobacco leaf curl Japan virus (TbLCJV) isolates from infected tomato (Lycopersicon esculentum) plants in Nara (-[Jp2], 2764nt; -[Jp3], 2761nt), Kochi (-[Koc], 2760nt) and Yamaguchi (-[Yam], 2758nt) Prefectures, of Japan were determined. These sequences were compared with each other and the sequences of further begomoviruses from Japan. TbLCJV, TbLCJV-[Jp2], TbLCJV-[Jp3], TbLCJV-[Koc], TbLCJV-[Yam], Honeysuckle yellow vein mosaic virus (HYVMV), Eupatorium yellow vein virus (EpYVV), EpYVV-[MNS2], EpYVV-[SOJ3], EpYVV-[Yam] and EpYVV-[Tob] are monophyletic. The intergenic region (IR) of TbLCJV has highest nucleotide sequence identity with that of HYVMV (93%) whereas the rest of the genomic DNA had higher identity with that of TbLCJV-[Jp2] or -[Jp3] (91100%) than with that of HYVMV. In conclusion, TbLCJV has a chimeric genome which may have arisen by recombination between TbLCV-[Jp2] or -[Jp3]-like and HYVMV-like ancestors. Similarly, TbLCJV-[Yam] DNA has a hybrid genome, with a major parent HYVMV and minor parent TbLCJV-[Koc].     

A temperature-dependent inhibitory activity of serum on the capacity of Saccharomyces cerevisiae-derived hepatitis B surface antigen to bind to monocytes

Summary. Hepatitis B surface antigen, when produced in yeast (rHBsAg), is capable of binding to cells that express the lipopolysaccharide coreceptor CD14. This interaction is enhanced by a serum protein, the lipopolysaccharide binding protein (LBP). Here we report that most of the rHBsAg particles that attached to monocytes at 0°C, were not endocytosed but were released back into the serum-containing binding buffer at 37°C. Additionally, serum-dependent binding at 37°C was weak when compared to the serum-dependent attachment at 0°C. Pre-incubation at 37°C of cells together with serum did not abolish binding of freshly added rHBsAg at 0°C. However, pre-incubation of rHBsAg with serum at 37°C reduced attachment to cells following incubation at 0°C. Soluble CD14 and LBP, two serum proteins which can act as phospholipid transfer molecules, were shown not to be responsible for the inhibitory effect. Pre-incubation at 37°C of rHBsAg in serum-free hepatoma cell line-conditioned media resulted in a pronounced reduction in subsequent binding to cells at 0°C. These observations suggest that the temperature-dependent inhibitory effect is caused by serum factors that are probably secreted by hepatocytes.     

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.     

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.