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Biomedical subjects

M Kojima

Publications and source records attributed to M Kojima.

At least 19 recordsLinked to original sources

Effective blocking of natural cytotoxicity of young rabbit serum on murine thymocytes by high concentration of glucose in complement-dependent cytotoxicity method.

By an accidental observation, the natural cytotoxicity of 21-day-old rabbit sera to murine thymocytes was found to be almost completely inhibitable by 4.5 mg/ml glucose. This enabled us to use unabsorbed young rabbit serum as a source of complement for antibody-mediated cytotoxic treatment of murine thymocytes. This glucose-sensitive natural cytotoxic activity was recovered in the immunoglobulin fraction purified on an affinity column, and was dependent on complement activity. The unabsorbed 21-day-old rabbit sera had much higher complement activity than those from 14-day-old rabbits and commercially available low background complement. Indeed, complete depletion of CD4+ cells from murine thymocytes and splenocytes was achieved using unabsorbed 21-day rabbit serum as a source of complement and a monoclonal antibody GK1.5 which has been claimed to be inefficient in complement-dependent killing. Owing to the increase in complement titer and circulating blood volume 14-21 days after birth, the use of 21-day-old rabbit sera assures a much more abundant supply of complement for the treatment of murine lymphocytes.

Animals

Fulminant hepatitis related to transmission of hepatitis B variants with precore mutations between spouses.

A precore defective variant of hepatitis B virus has been indicated to cause fulminant hepatitis in various instances such as intrahospital outbreaks or mother-to-child transmission of hepatitis B virus. To learn whether similar variants are involved in interspouse transmission, we analyzed three cases of fulminant hepatitis B that developed in formerly healthy subjects whose only exposure to hepatitis B virus was contact with their longtime spouses, who were carriers of HBV and positive for antibody to HBe. The DNA clones for precore and S genes were propagated from patients and spouses and sequenced. Because of the conservation of S-gene sequences and the identity of subtypes between patient and spouse, it was suggested that patients were infected with hepatitis B virus from their spouses, not from other sources. A TGG-to-TAG mutation at the 28th codon of the precore gene of hepatitis B virus was commonly observed in all DNA clones from patients with fulminant hepatitis and from their spouses. A 29th-codon GGC-to-GAC mutation was additionally evident in DNAs from one patient-and-spouse couple. A significant rise in the circulating hepatitis B virus concentration was transiently observed in the index spouse of this case just before development of fulminant hepatitis in her husband. The increase in circulating HBV DNA was associated with a rise in abundancy of variants with mutations at both the 28th and 29th codons, compared with variants with only a 28th-codon mutation. The double mutation in hepatitis B virus DNA may either help the virus escape immune surveillance or replicate at a higher rate than before.

Base Sequence

Hepatitis B surface antigen particles of subtypes adw and adr, and compound subtype (adwr) in symptom-free carriers in Japan.

Of sera from 1,878 Japanese blood donors who carried hepatitis B surface antigen (HBsAg), 420 were subtyped as adw (22.4%) and 1,443 as adr (76.8%); only 15 (0.8%) contained HBsAg of subtype ayw or ayr. Sera with HBsAg/adr had higher HBsAg titres than those with HBsAg/adw (geometric mean of haemagglutination titre: 10.1 +/- 2.4 vs. 9.7 +/- 2.4, p less than 0.01), and a higher prevalence of hepatitis B e antigen (24% vs. 13%, p less than 0.001). Carriers of HBsAg/adr progressively predominated over those of HBsAg/adw with increasing age. Of sera from 1,863 carriers of HBsAg/adw or HBsAg/adr, 182 (9.8%) contained HBsAg particles with both subtypic determinants in the w/r allele. The presence of w and r determinants on the same particles was ascertained by sandwiching them between monoclonal antibody with the specificity for w and that with the specificity for r. HBsAg particles of compound subtype (adwr) were found more often in sera with hepatitis B e antigen than those without it (145/403 [36.0%] vs. 37/1,460 [2.5%], p less than 0.001). Sera with HBsAg/adwr particles had HBsAg titres higher than those without them (12.4 +/- 1.9 vs. 9.7 +/- 2.3, p less than 0.001). HBsAg/adwr particles arise from phenotypic mixing of the S-gene product of wild-type virus and that of mutants with point mutations for subtypic changes. The results obtained indicated that HBV strains of subtype adr have a higher replicative activity than those of adw, and suggested that mutations in the S gene for subtypic changes would be associated with an active replication of hepatitis B virus.

Adult

Outward permeability of the blood-retinal barrier.

To characterize quantitatively the active transport mechanism of the blood-retinal barrier (BRB), we estimated the inward (Pin) and outward permeability (Pout) of the BRB in monkey eyes using vitreous fluorophotometry. Pin values for fluorescein (F) and fluorescein glucuronide (FG) were simulated by computer in separate experiments following measurements of intraocular fluorescence at 1 h following the intravenous injection of F or FG. The estimated mean Pin values for F and FG were 4.7 +/- 1.6 and 5.9 +/- 1.7 x 10(-6) cm/min, respectively, in ten eyes of five monkeys. At 1 month thereafter, F was injected into the right vitreous cavity and the same amount of FG was injected into the left vitreous cavity. The intraocular kinetics of fluorescence were measured at 6-24 h postinjection. As estimated by the simulation model, the mean Pout values were 7.7 +/- 2.6 and 1.7 +/- 0.9 x 10(-4) cm/min for F and FG, respectively. The Pout/Pin ratio was 160 +/- 78 for F and 26 +/- 9 for FG. The intraperitoneal injection of probenecid resulted in a significant decrease in the Pout value for F (P less than 0.005) but had no significant effect on that for FG, suggesting that F is excreted from the retina via an active transport mechanism; as characterized by the Pout/Pin ratio, the actual magnitude of the latter is far greater than that previously reported.

Animals

A case of treatment-related leukoencephalopathy: sequential MRI, CT and PET findings.

A case of treatment-related leukoencephalopathy is presented. A patient with medulloblastoma was postoperatively treated with craniospinal axis irradiation. One month after irradiation, weekly intrathecal administration of methotrexate was performed 4 times to treat cerebrospinal fluid dissemination of the tumor. Two months after the initiation of intrathecal chemotherapy, the patient became somnolent and developed decerebrate posturing. Magnetic resonance imaging showed diffuse leukoencephalopathy. Positron emission tomography revealed a diffuse decrease in glucose uptake in the deep white matter. Auditory evoked potential also showed diffuse abnormalities, not only in the cerebrum, but also in the brain stem. High dose intravenous leucovorin rescue was attempted without any neurologic improvement.

Blood Proteins

Intravesical instillation of adriamycin for bladder tumors.

In the present series of trials, Adriamycin (ADM) intravesical instillation therapy was found to be effective against tumors of papillary morphology measuring less than 10 mm in diameter that were of low pathological stage and low histological grade. The rate of complete disappearance increased in proportion to the concentration of ADM and the duration of retention of the drug, although the efficacy rates were almost the same in each trial. On the other hand, side effects on the bladder were reduced when the instilled solution was lower in concentration and the retention time was short.

Administration, Intravesical

On the variability of the 3' terminal sequence of the turnip mosaic virus genome.

The sequence of the 3'-terminal 1223 nucleotides (nts) of a Japanese isolate of turnip mosaic virus (TuMV-Jap) RNA has been determined. The sequence reveals a single open reading frame (ORF) which terminates at a position 212 nts upstream of the 3' poly(A)-tract. Determination of the N-terminal amino acids of TuMV-Jap coat protein (CP) mapped the CP cistron within this ORF and revealed a Glu-Ala dipeptide sequence as the putative cleavage site by which the CP is released from the viral polyprotein. The predicted amino acid sequence of the TuMV-Jap CP shows 97.2% identity with that of a Canadian isolate of TuMV (TuMV-Can) and 99% with a second, Chinese, isolate (TuMV-Chi). However, the 3'-terminal non-translated region (NTR) of TuMV-Jap RNA is significantly shorter (212 nts) than the 3'-NTR of TuMV-Can RNA (668 nts), but of equal length as the 3'-NTR of the TuMV-Chi isolate which also measures 212 nts. The 3'-NTRs of both the TuMV-Jap and TuMV-Chi RNAs show homology with the first 201 nucleotides of the TuMV-Can RNA 3'-NTR. A search in the EMBL nucleotide sequence database revealed that the 467 nt-long unique extension of the 3'-NTR of TuMV-Can RNA has 89.8% homology to a part of the chloroplast ribosomal protein 12 gene (rsp12-gene). Irrespective of the origin of this extra sequence in the reported TuMV-Can sequence, which may have been introduced by a genuine RNA recombination event, it is concluded that the standard TuMV genome has a CP gene of 864 nts and an conserved 3'-NTR of approximately 212 nucleotides in length.

Amino Acid Sequence

Emperipolesis of lymphoid cells by human follicular dendritic cells in vitro.

Isolated follicular dendritic cells (FDCs) showed true and pseudoemperipolesis of fresh tonsillar lymphocytes, even after long-term (50-day) cultivation. Emperipolesis by FDCs was not restricted by allotype specificity, nor was it inhibited by the addition of antibodies against MHC-I & II antigens. Follicular dendritic cells predominantly engulfed B-cells; monocytes and macrophages were not found between FDC cytoplasmic extensions. When highly purified T-cell populations were added to FDC cultures emperipolesis of T-cells occurred, particularly those of the CD4-positive phenotype. Mitoses appeared within 6 h in the emperipolesed lymphocytes and, after an additional 18 h, some lymphocytes exhibited apoptosis.

Antigens, Differentiation

Genetic drift of hepatitis C virus during an 8.2-year infection in a chimpanzee: variability and stability.

Extensive variability in genomic sequence, especially at "hypervariable regions" within the NS1/E2 region of the long open reading frame, has been reported for RNA cloned from hepatitis C virus (HCV)-infected humans and chimpanzees. However, genetic changes of HCV occurring during the course of chronic infections in humans and animals have been evaluated only for partial sequences of the HCV genome. We compared two full-length cDNA sequences of HCV obtained from a chimpanzee that was experimentally infected with the HC-J4 strain of HCV: one during the early acute phase and another during a chronic phase 8.2 years afterward. Both isolates had 9412 nucleotides plus the 3' poly(U) tail with varying length organized as follows: 5'UTR (1-341); C (342-914); E (915-1490); NS1/E2 (1491-2528); NS2 (2529-3359); NS3 (3360-5186); NS4 (5187-6380); NS5 (6381-9371); and 3'UTR (9372-9412). We found that 111 (1.18%) of the 9412 nucleotides differed between the two isolates and estimated the mutation rate as approximately 1.44 x 10(-3) base substitutions per site per year. Changes in amino acid coding were associated with 42 mutations, 8 of which were clustered at 5' end of NS1/E2 coding region, so-called "HVR-1." We analyzed the HVR-1 and HVR-2 sequences during the course of infection and found that homologous populations were present at the beginning of infection, and sequence heterogeneity within the region had developed 3.5 years later. Two regions of the HCV genome were characterized by a high degree of conservation of nucleotide sequence: 5'UTR and the 3' half of the NS4 region. The possible secondary structure of the 5'UTR suggests a region for internal ribosomal entry. The 3' half of the NS4 region may also have some specific function which depends upon a strict conservation of nucleotide sequence.

Amino Acid Sequence

Endovascular coil embolization of a recurrent giant internal carotid artery aneurysm via the posterior communicating artery after cervical carotid ligation: case report.

The case of a 29-year-old man with a giant fusiform aneurysm of the left internal carotid artery (ICA) is presented. The aneurysm, treated by cervical ICA ligation and extracranial-intracranial bypass, recurred 4 years later owing to recruitment of the posterior communicating artery (PCoA). Because of the previous bypass surgery a direct surgical approach was excluded. After an initial failure with balloon embolization, the aneurysm was embolized successfully with occlusive platinum microcoils through the microcatheter navigated into the aneurysm via the enlarged PCoA. Endovascular coil embolization may be useful in the treatment of cerebral aneurysms not amenable to direct surgery or balloon embolization.

Adult

Nerve growth factor (NGF)-mediated up-regulation of low-affinity NGF receptor gene expression in cultured basal forebrain cholinergic neurons from postnatal 3-day-old rats.

We examined the effect of nerve growth factor (NGF) on the expression of low-affinity NGF receptor (LNGFR) in cultured P3 basal forebrain cholinergic neurons, on which NGF acts to promote differentiation. Based on the results of the RT-PCR (reverse transcriptase-polymerase chain reaction) method, over a 2-fold increase in the LNGFR mRNA level was found in NGF-treated cultures compared with control cultures at one day after the addition of 100 ng/ml NGF. This increase was maintained for up to 3 days after the addition of NGF. The increase in LNGFR mRNA was found even at 1 ng/ml NGF (= 40 pM), indicating that the up-regulation of the LNGFR mRNA level in cultured cholinergic neurons occurred at an NGF concentration near the Kd value for the high-affinity NGF receptor. In addition, immunohistochemical staining showed stronger staining with a monoclonal anti-LNGFR antibody of the cholinergic neurons in NGF-treated cultures than those in control cultures. Our results suggest that NGF can up-regulate LNGFR expression at the mRNA and protein levels in cultured P3 basal forebrain cholinergic neurons and that this mechanism may play an important role in potentiating the effect of NGF on these neurons.

Acetylcholine

Differences in DNA damage induced by mutagenic and nonmutagenic 4-aminoazobenzene derivatives in Escherichia coli.

DNA lesions produced in Escherichia coli AB2500 (uvrA) exposed to the carcinogen N-hydroxy-3-methoxy-4-aminoazobenzene (N-OH-3-MeO-AAB) or the noncarcinogen N-hydroxy-2-methoxy-4-aminoazobenzene (N-OH-2-MeO-AAB) were investigated by alkaline sucrose gradient sedimentation and 32P-postlabeling analysis. Alkali-labile sites appeared to be formed equally in cells treated with both aminoazobenzene derivatives. 32P-Postlabeling analysis revealed that the 3-MeO-AAB-DNA adduct level was 25-fold higher than that for 2-MeO-AAB-DNA adducts. In addition to major adducts, 4 minor spots were detected in N-OH-3-MeO-AAB-treated cells, while only one major adduct was found in N-OH-2-MeO-AAB-treated cells. The mutagenicities and cytotoxicities were also determined with E. coli with different repair capacities; we found that repair of 3-MeO-AAB damages is strongly dependent on the UVR repair system. Moreover, N-OH-3-MeO-AAB, but not N-OH-2-MeO-AAB, could induce recA and umuC gene expression, which was higher in uvrA strains than in the wild type.

Bacterial Proteins

Malignant schwannoma with rhabdomyoblastic and melanocytic differentiation.

This is the first case report of an unusual malignant tumor with components of malignant schwannoma, rhabdomyosarcoma, and melanoma, occurring in an adult. The tumor was observed in the left temple in a 35-year-old man without von Recklinghausen's disease. The possible derivation of this tumor from primitive neuroectodermal cells is discussed.

Adult

Electrotransfection of protoplasts from tomato, wild tomato, barley and chrysanthemum with tobacco mosaic virus RNA.

Protoplasts isolated from tomato, wild tomato, barley and chrysanthemum were electrotransfected with tobacco mosaic virus (TMV) RNA under almost the same optimum electric conditions: five square DC pulses of 50 microseconds duration at 500 to 800 V/cm, with the protoplasts suspended at 2 x 10(5)/ml in 0.5 M-mannitol containing 100 microM-MgCl2 and 10 to 20 micrograms/ml TMV RNA. ELISAs of these transfected protoplasts showed that the yields and the growth curves of the virus were quite similar, indicating a lack of host specificity in the initially infected cells of these plants.

Chrysanthemum cinerariifolium

Immunological detection and quantitation of DNA adducts formed by 4-aminoazobenzene species in vivo.

Antibodies to 3-methoxy-4-aminoazobenzene (3-MeO-AAB) and 2-methoxy-4-aminoazobenzene (2-MeO-AAB) DNA adducts were raised in rabbits against in vitro-adducted DNA samples. The enzyme-linked immunosorbent assay (ELISA) was used to determine the sensitivity and specificity of these antibodies. They proved highly specific for the modified DNA used as the immunogen, but cross-reacted with each other. Moreover, they showed cross reactivity with DNA modified by 4-(o-tolylazo)-o-toluidine, but not by other carcinogens, such as 4-aminobiphenyl or 4-nitroquinoline 1-oxide. The 50% inhibition level of antibody binding in the competitive ELISA was at 10-20 fmol of modified base per assay (equivalent to 1-2 adducts per 10(6) bases). Immunohistochemical staining indicated that these antibodies bind specifically to nuclear components of the liver in rats given either 3-MeO-AAB or 2-MeO-AAB at the dose of 50 mg/kg body weight.

Animals

Malignant lymphomas of the nasal cavity and paranasal sinuses. A clinicopathologic and immunohistochemical study.

The clinicopathologic and immunohistological features of 20 Japanese patients with non-Hodgkin's lymphomas (NHLs) limited to the sinonasal area were studied using a broad panel of T- and B-cell markers on paraffin-embedded and fresh frozen tissue. All cases showed a diffuse growth pattern. Nine cases were B-cell lymphomas (immunoblastic n = 4, centroblastic n = 3, immunocytoma n = 1, centrocytic n = 1), and nine were T-cell lymphomas (pleomorphic medium and large cell n = 8, angioimmunoblastic n = 1). In two cases, the cell lineage could not be determined. No morphologic features of angiocentric/angiodestructive lymphoproliferative lesions or lymphoepithelial lesions in ductal or glandular epithelium were seen in our series. Eight (89%) of the nine T-cell tumors and four (44%) of the nine B-cell neoplasms involved both the nasal cavity and paranasal sinuses. Six of the nine T-cell neoplasms showed a clinical presentation of rhinitis, whereas all of the B-cell neoplasms showed tumor masses in the nasal cavity and/or paranasal sinuses. The two-year survival rate for T-cell lymphomas was poorer than that for B-cell lymphomas. The five-year survival of patients with NHLs involving both the nasal cavity and paranasal sinuses was also poorer than that of patients in whom NHLs were limited to the nasal cavity.

Adult

Blood-ocular barrier permeability in monkeys.

The permeability of the blood-ocular barrier was investigated in five monkeys using vitreous fluorophotometry (VFP). Inward permeability (Pin) of the blood-retinal barrier was calculated by a computer simulation method. Kinetic VFP was performed after intravitreal injection of fluorescein (F) or fluorescein monoglucuronide (FG). The estimated mean value of Pin (x10(-6) cm/min) was 4.8 (SD 1.2). The mean rates of loss (per hour) of F from the anterior chamber (Ka) and the vitreous (Kv) were 0.11 (SD 0.01) and 0.13 (SD 0.03), respectively, which were approximately three and four times greater than those of FG (0.04 (SD 0.01) and 0.03 (SD 0.01), respectively). Probenecid administered intraperitoneally decreased both the Ka and the Kv of F significantly but had no effect on the Ka or the Kv of FG, suggesting that F was excreted from the eye with the aid of the active transport mechanism. The results of comparative studies of the rates of loss of F from the anterior chamber (Ka) and from the vitreous (Kv) suggested that active transport was more predominant in the blood-retinal barrier than in the blood-aqueous barrier.

Animals

Role of endogenous ANP in sodium excretion in rats with experimental pulmonary hypertension.

To explore the roles of endogenous atrial natriuretic peptide (ANP) in the volume regulation, we examined effects of antiserum for ANP or a neutral endopeptidase inhibitor, thiorphan, in rats with monocrotaline (MCT)-induced pulmonary hypertension. ANP concentrations in the plasma and right ventricle and ANP mRNA in the right ventricle of MCT-treated rats were higher than in vehicle-treated rats. The administration of the ANP antiserum into the MCT-treated rats did not affect the right atrial pressure or blood pressure but significantly decreased urinary excretion of Na by 60%. No decrease occurred in the control rats. Thiorphan dose dependently increased the urinary excretion of Na by 140% without influencing the right atrial pressure or blood pressure. This natriuresis was associated with 50 and 450% increases in ANP concentrations in the plasma and urine, respectively. The degrees of increases in urinary Na excretion, ANP, and guanosine 3',5'-cyclic monophosphate were significantly greater in the MCT-treated rats than in the control rats. Thus an increased secretion of ANP in pulmonary hypertension actually contributes to Na excretion. The augmentation of endogenous ANP activity may further potentiate the compensatory role of this peptide in the regulation of body fluid volume.

Animals