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Biomedical subjects

A Adachi

Publications and source records attributed to A Adachi.

At least 37 records · Page 2Linked to original sources
PubMed2001

Analysis of the cell-dependent replication potentials of human immunodeficiency virus type 1 vif mutants.

Eleven in-frame vif gene mutants of HIV type 1 produced in non-permissive cells were examined for their replication potentials in various CD4-positive and -negative cell lines. Virus replication for each mutant was monitored by using several single- and multiple-cycle infectivity assays. Except for a mutant with wild-type phenotype, most mutants were severely defective for replication in all the cell lines as expected from the producer cell-dependent functioning of Vif so far reported. In contrast, two mutants, which have mutations in the hydrophilic or effector regions of Vif were found to have target cell-dependent replication potentials. These results demonstrate the presence of a novel category of the vif mutants important for elucidation of the Vif function.

Amino Acid Sequence↗
PubMed2001

Lack of apoptosis in Sendai virus-infected HEp-2 cells without participation of viral antiapoptosis gene.

Sendai virus (SeV) has been reported to induce apoptosis in many types of cells. In HEp-2 cells, however, it did not induce apoptosis in most of the infected cells under the conditions in which vesicular stomatitis virus induced massive apoptosis. The use of a novel technique, which allows the detection of viral antiapoptotic activity in the infected cells, showed that SeV does not have any antiapoptotic activity to interfere with the induction of apoptosis. Consistently, vesicular stomatitis virus-induced apoptosis was not interfered with by preinfection with SeV. These results indicate that the observed lack of apoptosis in these SeV-infected cells does not result from the suppression of apoptosis by viral antiapoptotic activity in the infected cells and suggest that, without activating a signaling pathway for the induction of apoptotic response in the infected cells, SeV can escape apoptosis of the cells, allowing long-term survival of the infected cells.

Animals↗
PubMed2001

Efficiency of rice bran for removal of organochlorine compounds and benzene from industrial wastewater.

Rice bran was found to effectively adsorb several organic compounds, such as dichloromethane, chloroform, carbon tetrachloride, trichloroethylene, tetrachloroethylene, and benzene. Equilibrium adsorption isotherms conformed to the Freundlich type (log-log linear). The adsorption of dichloromethane and chloroform by rice bran was observed over the range of pH 1-11. Therefore, rice bran is applicable for treatment of wastewater over a wide pH range. Dichloromethane was successfully removed from water samples with an average removal efficiency of 70% after 60 min when rice bran was added to water samples containing from 0.006 to 100 mg/L dichloromethane. The removal of these organochlorine compounds and benzene by rice bran was attributed to the uptake by intracellular particles called spherosomes. Here, we report the results of a fundamental study of the efficiency of rice bran for removal of organochlorine compounds and benzene using a batch system on the laboratory scale, and describe elucidation of the mechanism of removal of these compounds by rice bran.

Adsorption↗
PubMed2001

Cyclophilin A-independent replication of a human immunodeficiency virus type 1 isolate carrying a small portion of the simian immunodeficiency virus SIV(MAC) gag capsid region.

Hybrid viruses between human immunodeficiency virus type 1 (HIV-1) and simian immunodeficiency virus strain mac (SIV(MAC)) are invaluable to various fields of HIV-1 research. To date, however, no replication-competent HIV-1 strain containing the gag capsid (CA) region of SIV(MAC) has been reported. To obtain the viable gag gene chimeric virus in an HIV-1 background, seven HIV-1 strains carrying a part of SIV(MAC) CA or a small deletion in the CA region were constructed and examined for their biological and biochemical characteristics. While all the recombinants and mutants were found to express Gag and to produce progeny virions on transfection, only one chimeric virus, which has 18 bp of SIV gag CA sequence in place of the region encoding the HIV-1 CA cyclophilin A (CyPA)-binding loop, was infectious for human cell lines. Although this chimeric virus was unable to grow in monkey lymphocytic cells like wild-type (wt) HIV-1 did, it grew much better than wt virus in the presence of cyclosporin A in a human cell line which supports HIV-1 replication in a CyPA-dependent manner. These results indicate that the transfer of a small portion of the SIV(MAC) CA region to HIV-1 could confer the CyPA-independent replication potential of SIV(MAC) on the virus.

Amino Acid Sequence↗
PubMed2001

Characteristic intracytoplasmic material in a perianal wart that contains human papillomavirus type 56 and 16 DNA.

We report the case of a perianal wart in which human papillomavirus type 56 and 16 DNAs were detected, arising in a 29-year-old healthy Japanese female. Clinically, moist erythema and small flat pinkish papules appeared around the anus and worsened over a period of 6 months. Histopathologically, no signs of dysplasia or atypia were apparent. An area of intracytoplasmic material, which was homogeneous and eosinophilic, was contained in each epidermal cell. The wart showed HPV-56 and HPV-16 DNAs by PCR and reverse-phase dot blot hybridization. Additional Southern blot analysis showed the amount of HPV-56 DNA to be more than 10 times that of HPV-16 DNA. This is the first case, to our knowledge, of a perianal wart with intracytoplasmic material, which contains HPV DNAs causing mucous lesions.

Adult↗
PubMed2001

Large-scale cDNA analysis of the maternal genetic information in the egg of Halocynthia roretzi for a gene expression catalog of ascidian development.

The ascidian egg is a well-known mosaic egg. In order to investigate the molecular nature of the maternal genetic information stored in the egg, we have prepared cDNAs from the mRNAs in the fertilized eggs of the ascidian, Halocynthia roretzi. The cDNAs of the ascidian embryo were sequenced, and the localization of individual mRNA was examined in staged embryos by whole-mount in situ hybridization. The data obtained were stored in the database MAGEST (http://www.genome.ad.jp/magest) and further analyzed. A total of 4240 cDNA clones were found to represent 2221 gene transcripts, including at least 934 different protein-coding sequences. The mRNA population of the egg consisted of a low prevalence, high complexity sequence set. The majority of the clones were of the rare sequence class, and of these, 42% of the clones showed significant matches with known peptides, mainly consisting of proteins with housekeeping functions such as metabolism and cell division. In addition, we found cDNAs encoding components involved in different signal transduction pathways and cDNAs encoding nucleotide-binding proteins. Large-scale analyses of the distribution of the RNA corresponding to each cDNA in the eight-cell, 110-cell and early tailbud embryos were simultaneously carried out. These analyses revealed that a small fraction of the maternal RNAs were localized in the eight-cell embryo, and that 7.9% of the clones were exclusively maternal, while 40.6% of the maternal clones showed expression in the later stages. This study provides global insights about the genes expressed during early development.

Animals↗
PubMed2001

Growth characteristics of SHIV without the vpu gene.

NM-3 is a prototype of chimeric virus between simian and human immunodeficiency viruses (SHIV). It grows in monkey lymphocytic cells in vitro and in vivo as well as in human cells. A mutant designated NM3-E65, which lacks expression of the entire vpu gene, was constructed from NM-3, and monitored for its replication property. Examination of growth properties in simian and human cells of SHIV, HIV, and their vpu mutants revealed that the vpu gene in the genome of the chimeric virus is functional, but non-essential for virus replication.

Animals↗
PubMed2001

[Audiological and equilibrium study of perilymphatic fistulas].

We clinically analyzed 15 cases of perilymphatic fistulas--11 caused by barotraumas and 4 idiopathic--identified by surgery between March 1995 and March 1999 at the Hyogo College of Medicine and affiliated hospitals. Subjects were 11 men and 4 women (aged 14 to 79 years (mean: 46.7 years)). All showed hearing loss in audiography and 12 cases reported tinnitus--stream-like in 5 and poping in 4. Dysequilibrium was seen in 9 cases. Perilymph leakage was detected intraoperatively from the oval window in 9, from the round window in 4, and from both windows in 1, while another had leakage from the fissura ante fenestram. After surgery, hearing level improved by over 10 dB in 9 of the 11 cases operated on within 14 days after onset. Hearing did not improve in 3 of 4 operated on later. Vertigo disappeared after surgery. Dizziness tended to persist in those having canal paresis or paralytic nystagmus before surgery. We suggest that patients with progressive hearing loss should be operated on as soon as possible and that patients with dysequilibrium or without response to conservative treatment undergo surgery within 14 days of onset.

Adolescent↗
PubMed2000

Activity of neurons in the nucleus of the solitary tract of rats: effect of osmotic and mechanical stimuli.

Patch-clamp recordings were used to examine the osmosensitivity and mechanosensitivity of neurons in the caudal part of the nucleus tractus solitarius in coronal slices from rat brain. Firing rates and membrane potentials were measured as slices were exposed to perfusate which varied in its osmolality and/or sodium concentration. In all cells tested, the responses to change in the sodium concentration of perfusate were duplicated by osmolality changes of sucrose or mannitol. When nucleus tractus solitarius cells were tested with changes in pressure applied via the pipette, responses to positive or negative pressure paralleled their responses to osmotic stimulation. We suggest that a mechanosensitive receptor exists on osmosensitive neurons within the nucleus tractus solitarius, and this receptor may be responsible for changes in the firing rate and membrane potential which occur in the nucleus tractus solitarius neurons.

Action Potentials↗
PubMed2000

Isolation and characterization of simian immunodeficiency virus variants that are resistant to nonnucleoside reverse transcriptase inhibitors.

Non-nucleoside reverse transcriptase inhibitors (NNRTIs) act quite specifically on human immunodeficiency virus type 1 (HIV-1). In general, they are not effective on human immunodeficiency virus type 2 (HIV-2) or simian immunodeficiency virus (SIV). Only SIV strains from African green monkeys are sensitive to several NNRTIs. Here we isolated NNRTI- and 3TC-resistant SIVagm variants. Viruses resistant to delavirdine contained V112I and M231I substitutions, while those resistant to 3TC contained a M 185I substitution. These amino acids are highly conserved in HIV-1, HIV-2, SIVmac and SIVagm, and the M184I (M185I in SIVagm) substitution was observed in 3TC-resistant HIV-1 and SIVmac. The roles of the observed mutations in NNRTI-resistance of SIVagm and HIV-1 were further confirmed by site-directed mutagenesis. The present results have provided a new insight into the common mechanism of sensitivity of HIV- 1 and SIVagm to NNRTIs.

Amino Acid Sequence↗
PubMed2000

Abnormal elastic system fibers in fibrotic human liver.

The network of elastic system fibers in human fibrotic liver was investigated by histological methods, immunohistochemical staining, and electron microscopy. Type III collagen was seen not only in regions of portal fibrosis but also in the sinusoidal wall. However, elastic system fibers were not found in the Disse space of the sinusoidal wall. Elastic system fibers including oxytalan, elaunin, and elastic fibers were found successively in the course of elastogenesis. A few normal oxytalan fibers and abnormal oxytalan fibers were observed in the periportal tracts. Few normal elaunin and abnormal elaunin fibers were observed in regions of portal fibrosis but not in the surrounding margin. Elastic fibers, only in scarce amounts, were observed around the portal veins in the case of chronic active hepatitis but not in acute hepatitis. Abnormal oxytalan fibers were seen as a bundle of wavelike microfibrils and had an irregular arrangement. Abnormal elaunin fibers were not associated with bundles of microfibrils. Abnormal elaunin fibers in large amounts were found interspersed with spiraled collagen, which most likely indicates that the oxytalan fibers degenerated in the course of elastogenesis. Thus, in a fibrotic liver it is possible that synthesis of normal elaunin and elastic fibers does not occur or that the quantity of such fibers synthesized may be small because of the effect of the degenerated oxytalan fibers. As a characteristic of liver fibrosis, the composition of abnormal elastic system fibers and spiraled collagen differs from that in other fibrotic organs.

Adult↗
PubMed2000

Elimination of HIV-1 plasmid DNA from virus samples obtained from transfection by calcium-phosphate co-precipitation.

Molecular genetics is a powerful tool to analyze the replication cycle of human immunodeficiency virus type 1 (HIV-1). Culture fluids obtained from HIV-1 plasmid-transfected cells by calcium-phosphate co-precipitation were treated with ethyleneglycol bis (beta-aminoethylether)-N,N,N',N'-tetraacetic acid (EGTA) and DNase I to obtain HIV-1 stocks virtually free of input plasmid DNAs. Even after amplification by polymerase chain reaction (PCR), no plasmid DNA was detected in cells following infection with EGTA/DNase I-treated virus samples. This method is particularly useful for the examination of the early replication phase of HIV-1 by PCR.

Calcium↗
PubMed2000

Requirements for presenilin-dependent cleavage of notch and other transmembrane proteins.

Ligand binding to receptors of the LIN-12/Notch family causes at least two proteolytic cleavages: one between the extracellular and transmembrane domains, and the other within the transmembrane domain. The transmembrane cleavage depends on Presenilin, a protein also required for transmembrane cleavage of beta-APP. Here, we have assayed the substrate requirements for Presenilin-dependent processing of Notch and other type I transmembrane proteins in vivo. We find that the Presenilin-dependent cleavage does not depend critically on the recognition of particular sequences in these proteins but rather on the size of the extracellular domain: the smaller the size, the greater the efficiency of cleavage. Hence, Notch, beta-APP, and perhaps other proteins may be targeted for Presenilin-mediated transmembrane cleavage by upstream processing events that sever the extracellular domain from the rest of the protein.

Amino Acid Sequence↗
PubMed2000

Characterization of apoptosis induced by sorbitol: a unique system for the detection of antiapoptotic activities of viruses.

The treatment of HEp-2 cells with sorbitol induced massive apoptosis rapidly. This method for inducing apoptosis is very useful to detect antiapoptotic activity of viruses as well as viral genes. Commitment to death occurred immediately upon incubation with sorbitol, even in the presence of pancaspase inhibitor, Z-VAD-FMK. Apoptosis is also induced by other polyhydric alcohols with more than four hydroxyl groups, but not induced by glycerol or ethylene glycol. Sorbitol treatment on ice did not induce apoptosis either. These results suggest that this induction of apoptosis does not result simply from high osmotic pressure but probably by the interaction of solutes through their physical nature (such as hydrophobicity) with the plasma membrane of the cells.

Amino Acid Chloromethyl Ketones↗
PubMed2000

Regulation of cell cycle and apoptosis by human immunodeficiency virus type 1 Vpr.

Biological effects of HIV-1 Vpr on CD4(+) cells were studied by an infection system. High-titered HIV-1 stocks pseudotyped with vesicular stomatitis virus G protein were prepared and used to inoculate into CD4(+ )T cells at high multiplicity of infection. Both cell- and virion-associated Vpr were demonstrated to arrest the cell cycle at the G2/M phase, and to induce cell apoptosis. Of note, morphologically apoptotic cells were shown to be arrested at the G2/M stage. No appreciable effect of Vpr on the anti-Fas antibody-mediated apoptosis was observed in this system.

Apoptosis↗
PubMed2000

Physiological significance of apoptosis in animal virus infection.

In contrast to insect viruses, animal viruses can produce considerable amounts of progeny virus in cells undergoing apoptosis. Nevertheless, viruses in general have acquired the ability to escape apoptosis of infected cells. These facts indicate that the role of apoptosis in virus infection is different in insect virus and animal virus, although both viruses need to avoid apoptosis of the infected cells for a viral life cycle in nature. In animal virus infection, the primary role of apoptosis is considered not to be a premature lysis of the infected cells (and the following abortion of virus multiplication) but to allow the dying cells to be phagocytosed by macrophages. This phagocytosis is able to prevent dysregulated inflammatory reactions at the site of virus infection and to initiate a specific immune response against the infected virus.

Animal Diseases↗
PubMed2000

Cell-dependent gag mutants of HIV-1 are crucially defective at the stage of uncoating/reverse transcription in non-permissive cells.

We have previously shown that some of the human immunodeficiency virus type 1 (HIV-1) gag matrix (MA), capsid (CA), and nucleocapsid (NC) mutants display host-cell-dependent replication potential, and that they are defective at the early phase of the virus replication cycle in non-permissive cells. To determine the defective replication stage of the cell-dependent mutants precisely, the processes of virus entry into cells and virus DNA synthesis were monitored by the highly sensitive enzyme-linked immunosorbent assay and polymerase chain reaction amplification analysis. The results obtained indicated that all the cell-dependent MA, CA and NC mutants are defective at the stage of uncoating/reverse transcription, and that a cellular factor(s) is involved in this process.

Capsid↗