Search PubMed⌕ Search

Biomedical subjects

K Horiuchi

Publications and source records attributed to K Horiuchi.

At least 127 records · Page 7Linked to original sources
PubMed1993

Detection of low copy numbers of Epstein-Barr virus by in situ hybridization using nonradioisotopic probes prepared by the polymerase chain reaction.

A highly sensitive in situ hybridization procedure was established using digoxigenin-11-dUTP-labeled probes that were prepared by the polymerase chain reaction (PCR). By using 12 sets of primers, BamHI-W fragment of the Epstein-Barr Virus (EBV) was amplified with labeled substrate in individual PCRs. Then the 12 probes (average size, 120 base pairs) were mixed and hybridized with cultured and RNase-treated Namalwa cells, which contain two copies of EBV genomes per cell. The strength of the signals was much stronger as compared with random-primed probe. Our results indicate that the size-averaged PCR probes magnified the sensitivity for detecting low copy numbers of virus genomes by in situ hybridization and that this technique has the potential for investigating latent virus infection in other clinical situations.

Base Sequence↗
PubMed1993

The occurrence of monocytoid B-lymphocytes in the spleen in gastric cancer.

Occurrence of monocytoid B-lymphocytes (MBL) in spleens of 34 patients with gastric cancer was examined. Histologic findings of gastric cancer including classification, depth of invasion, and stage of disease were defined based on the General Rules for the Gastric Cancer Study (Japan). MBL were defined morphologically as having abundant pale cytoplasm with distinct cell borders and small nucleus often with reniform shape. Immunohistochemically, these cells were B-cell in nature, i.e., CD15-, CD43-, CD45RA+, CD45Ro-, CD68-, CD74+, CDw75+, Mx-PanB(L26)+, MBI+, EMA-, PG-MI-. Clusters of MBL were found in 14 of 34 (41%) patients: they were found to be directly adjoining to the periarterial sheath or apart from the white pulp. In the cases without MBL, zonation of mantle zone and marginal zone was apparent with distinct secondary follicles in 72% of cases. Meanwhile, in spleens with MBL, the mantle zone showed atrophy in a half of cases, resulting in indistinct zonation of the mantle zone and marginal zone. Secondary follicles were distinct in less than 30% of cases. Correlation of the occurrence of MBL, evaluated by Spearman's correlation coefficient, revealed that the age was the most important factor (R = -0.4852, P = 0.00364): the median age of patients with and without MBL was 75.7 and 61.7 yr, respectively. The MBL were increased in gastric cancer as compared with other cancers (P < 0.03) and with noncancer spleens (P < 0.1). The age of gastric cancer patients with MBL was older than those in other cancers and noncancer patients. Therefore occurrence of MBL in spleen might be a function of aging.

Aged↗
PubMed1993

[Participation of environmental chemicals in experimental allergic conjunctivitis].

The effects of organophosphorus pesticides (trichlorfon and fenitrothion) and an organochlorine herbicide (paraquat) on experimental allergic conjunctivitis were studied quantitatively. Guinea pigs were passively immunized with antiserum to Japan cedar pollen. The above chemicals were administered subcutaneously 2 days prior to challenge with cedar pollen in the conjunctival sac, resulting in allergic conjunctivitis. The intensity of the conjunctival allergic reaction was quantitatively estimated by measuring the extravasation of Evans blue in the conjunctiva which had previously been administered intravenously. The allergic conjunctivitis was apparently aggravated by extremely low dosage level of organophosphorus pesticide (10(-4) approximately 10(-5) mg/kg) and organochlorine herbicide (10(-3) approximately 10(-4) mg/kg). The most intensive aggravating reaction was obtained 10(-5) mg/kg of organophosphorus pesticide and at 10(-4) mg/kg of paraquat dichloride. The aggravating effects of organophosphorus pesticide and organochlorine herbicide persisted at least two weeks after the single administration.

Animals↗
PubMed1993

[Usefulness and safety of dobutamine stress thallium myocardial scintigraphy].

Pharmaceutical stress test is performed for diagnosis of coronary artery disease in patients who can not perform exercise test. We studied whether high dose dobutamine stress test is useful and safe. Subjects consisted of 133 patients (pts) in doubutamine stress test and 45 pts in exercise test. Dobutamine stress test was initiated by 10 micrograms/kg/min, followed by 10 micrograms/kg/min increment every 3 minutes. Maximal doubutamine dose was 40 micrograms/kg/min for 3 minutes. Endpoints of dobutamine stress were as follows: ST depression equal to or more than (> or =) 0.2 mV: 30 pts, > or = 75% predictive maximum heart rate: 20 pts, > or = 20 mmHg systolic blood pressure fallen: 10 pts, 40 micrograms/kg/min for 3 minutes: 10 pts, chest pain: 7 pts, pressure rate product (PRP) > or = 25,000: 1 patient. Side effects of dobutamine stress test were as follows: palpitation: 52%, arrhythmia: 13%, angina and dyspnea: 7%, headache: 6%. These symptoms were disappeared within 2 minutes. One patient, who performed dobutamine test 2 day after PTCA, had myocardial infarction. At peak of dobutamine stress test, systolic blood pressure, heart rate and PRP were increased similar to exercise test. However, diastolic blood pressure was not changed. The concentration at peak of dobutamine was 25.5 +/- 7.8 micrograms/kg/min. The duration of dobutamine stress test was 7.3 +/- 2.3 minutes. The 65 patients (49%) need more than 20 micrograms/kg/min of dobutamine which was maximal loading dose in previous reports. For diagnosis of coronary artery disease, sensitivity was 91% (31/34) and specificity was 70% (7/10). From these data, high dose dobutamine stress test is useful and relatively safe stress method for diagnosis of coronary disease. This stress test would be performed in patients who can not perform exercise test.

Adult↗
PubMed1993

Quantification of the cross-link pentosidine in serum from normal and uremic subjects.

Pentosidine is a fluorescent cross-link compound that accumulates in human tissues from uremic and diabetic patients. Using SP-Sephadex C-25 pretreatment before reversed-phase HPLC, we developed a method for quantifying pentosidine in the acid hydrolysate of serum. We examined concentrations of pentosidine in serum from 98 patients with end-stage renal disease requiring hemodialysis and from 33 normal control subjects. The mean (+/- SD) concentration of pentosidine was significantly greater in serum from uremic patients than that from control subjects (1267 +/- 695 nmol/L vs 77 +/- 40 nmol/L, P = 0.0001). There was a significant correlation between serum pentosidine concentrations and age in control subjects (r = 0.453, P < 0.01), but not in uremic patients. Serum pentosidine significantly increased with the duration of hemodialysis treatment (r = 0.272, P < 0.05). The greater pentosidine concentrations in uremic patients may be caused by the increased synthesis of pentosidine (perhaps because of the retention of pentosidine precursors), by the retention of pentosidine molecule itself in end-stage renal failure, or both.

Adolescent↗
PubMed1992

Construction of a microphage variant of filamentous bacteriophage.

The intergenic region in the genome of the Ff class of filamentous phage (comprising strains fl, fd and M13) genome constitutes 8% of the viral genome, and has essential functions in DNA replication and phage morphogenesis. The functional domains of this region may be inserted into separate sites of a plasmid to function independently. Here, we demonstrate the construction of a plasmid containing, sequentially, the origin of (+)-strand synthesis, the packaging signal and a terminator of (+)-strand synthesis. When host cells harboring this plasmid (pLS7) are infected with helper phage they produce a microphage particle containing all the structural elements of the mature, native phage. The microphage is 65 A in diameter and about 500 A long. It contains a 221-base single-stranded circle of DNA coated by about 95 copies of the major coat protein (gene 8 protein).

Bacteriophage M13↗
PubMed1992

A single amino acid substitution reduces the superhelicity requirement of a replication initiator protein.

The origin of rolling circle replication in filamentous coliphage consists of a core origin that is absolutely required and an adjacent replication enhancer sequence that increases in vivo replication 30 to 100-fold. The core origin binds the initiator protein (gpII) which either nicks or relaxes negatively superhelical replicative form DNA (RFI). Nicking at the origin, but not relaxation, leads to initiation of DNA replication. Our results indicate that the ratio of nicking to relaxation (nicking-closing) in vitro depends on the superhelical density of the substrate. We have studied the effect of a single amino acid substitution in gpII, which allows wild-type levels of replication in the absence of the enhancer, on origin nicking and binding. The enhancer-independent mutation yields more nicking and less relaxation of RFI, compared to the wild-type protein. The mutant gpII also shows a reduced requirement for superhelicity of the substrate in the nicking reaction. At the same time, the mutant gpII increases the cooperativity of protein-protein interactions in origin binding. We propose that the relaxation activity of gpII negatively regulates replication initiation, and that both increase in the negative superhelicity of the substrate and action of the replication enhancer may antagonize the relaxation activity.

Amino Acid Sequence↗
PubMed1992

Vascularized fibular graft for reconstruction after resection of aggressive benign and malignant bone tumors.

From 1982 to 1991, 19 patients at Nara Medical University, Kashihara, Japan, underwent resection of aggressive benign and malignant bone tumors, with limb salvage and reconstruction by free vascularized fibula grafts. The patients were followed up for an average of 54 months. The reconstructed site was the jaw in 6 cases, upper extremity in 3, spine in 1, pelvis in 2, and lower extremity in 7. Six patients had aggressive benign lesions, and 13 had malignant lesions. The sizes of the resultant bone defect ranged from 6 to 20 cm, and the lengths of fibular used ranged from 8 to 24 cm. The average time to union was 4.1 months (2-9 months) in the extremities and pelvis. Local recurrence was observed in 3 cases, for whom vascularized fibula grafts were performed for recurrent tumors. In cases of primary untreated tumors, no recurrences occurred. Therefore, this procedure should be performed at the time of primary operation after extensive resection of an aggressive benign or malignant bone tumor.

Adolescent↗
PubMed1992

Malignant lymphoma of the liver. Report of five cases and review of the literature.

Primary hepatic lymphoma is a rare disease. We report five cases here and summarize clinical and pathologic features of our own and reported cases from Western countries and Japan. The total number of cases was 68. The age of patients ranged from 7 to 87 years (median 55) with a male-to-female ratio of 3.1:1. Chronic hepatitis or cirrhosis before onset of hepatic lymphoma was noted in 44% of Japanese cases and 9.6% of Western cases. Macroscopically, the liver was occupied by solitary mass (60%), multiple masses (35%), or a diffuse lesion without nodule formation (5%). Histologically all cases were non-Hodgkin's lymphoma with the diffuse large cell type being most common. Three cases (4.4%) were follicular lymphoma. Immunohistochemically about 80% of the cases were B-cell type. Follow-up study showed that hepatic lymphoma had a relatively favorable prognosis when early detection of the disease was possible.

Female↗
PubMed1992

Clinical trial of a lyophilized inactivated hepatitis A candidate vaccine in healthy adult volunteers.

The safety and immunogenicity of a lyophilized inactivated hepatitis A vaccine was tested in healthy adult male volunteers. Thirty-six volunteers, all of whom were negative for antibody to HAV (anti-HAV), were divided into three dosage groups, 1.0, 0.5 and 0.25 micrograms of viral protein, respectively. Each group received a total course of three intramuscular injections at months 0, 1 and 6. Slight side effects were noted after 16 of 99 injections and the occurrence and degree were almost identical to those of other commercial vaccines. On the other hand, all subjects had measurable titres of serum anti-HAV neutralizing antibodies as early as 2 months after the first injection. The mean values of serum anti-HAV neutralizing antibodies at 7 months in the 1.0, 0.5 and 0.25 micrograms dose groups were 64-, 12-, and 9-fold higher, respectively, than those observed at 5 days in five recipients given 7.5 mg kg-1 body weight of immune serum globulin (ISG).

Adolescent↗
PubMed1992

Quantitative analyses of urinary pyridinoline and deoxypyridinoline excretion in patients with hyperthyroidism.

It is well established that bone turnover increases and bone resorption exceeds bone formation in patients with hyperthyroidism. Recently, urinary pyridinoline and deoxypyridinoline have been employed as indicators of bone resorption. In this study, urinary pyridinoline and deoxypyridinoline levels in fourteen female patients with untreated hyperthyroidism were measured and compared with levels in 134 healthy females. Values of urinary pyridinoline and deoxypyridinoline were significantly higher than in age-matched controls. Moreover, values of serum free triiodothyronine correlated with urinary pyridinoline (r = 0.769, p < 0.01), and urinary deoxypyridinoline (r = 0.799, p < 0.001) in patients with hyperthyroidism.

Adult↗
PubMed1992

Radioreductive effect of bestatin (Ubenimex) in BALB/c mice.

The radioreductive effect of Bestatin (Ubenimex), a biological response modifier, was examined in BALB/c mice. Consecutive daily administrations from day 0 to day 3 after lethal irradiation resulted in a dose reduction factor (DRF) of 1.03. In haematological studies Bestatin enhanced the recovery of peripheral leucocytes in sublethally irradiated mice by activating colony-stimulating factors (CSF) in serum, leading to enhanced recovery of GM-CFC. There was no enhancement of CSF in BALB/c nude mice. On the other hand, Bestatin showed no effect on day 12 CFU-S and day 7 CFU-S after the sublethal irradiation, though there was a trend that the drug stimulated numbers of day 7 CFU-S after irradiation. These results strongly suggest that Bestatin enhanced the CSF activity by modulating T cell function, resulting in the augmentation of peripheral leucocytes and prolongation of survival of irradiated mice. This drug now used in the clinic proved to be modestly effective.

Acute Disease↗
PubMed1992

Malignant lymphoma of the breast. Immunologic type and association with lymphocytic mastopathy.

Clinical and pathologic findings in 19 cases of primary non-Hodgkin's lymphoma of the breast collected from several hospitals in Japan were reviewed. All patients were women (median age, 45 years) and they usually had breast masses that had recently become enlarged. The sites of the lesions were the right breast in eight cases, the left breast in eight, and both breasts in one. The locations of two masses were unknown. Lymphoma recurred in the opposite breast in three cases 14, 23, and 23 months after surgery. Histologically, diffuse large cell lymphoma was the most common form of disease (63%). One lesion was a follicular lymphoma. The so-called lymphoepithelial lesion, a characteristic finding for mucosa-associated lymphoid tissue type lymphomas, was observed in eight cases (42%). Immunohistochemical analysis revealed that all but two tumors were of B-cell type; such findings confirmed morphologically based conclusions. Histologic and immunohistochemical evidence of lymphocytic mastopathy, a recently described autoimmune disease of the breast, was found in most of the cases. Formation of lymphoid follicles in or around the tumors was found in five cases (26%). Based on these findings, it is suggested that most mammary lymphomas are B-cell tumors and they may be associated with coexisting or antecedent lymphocytic mastopathy.

Adult↗
PubMed1992

Immunochemical studies and complete amino acid sequence of the streptokinase from Streptococcus pyogenes (group A) M type 12 strain A374.

The complete amino acid sequence of the streptokinase (SKase) of Streptococcus pyogenes M type 12 strain A374, isolated from a patient with poststreptococcal glomerulonephritis (PSGN), was determined. The epitope domain for the monoclonal antibody N-59, which cross-reacts with SKases of both the PSGN-associated strain and S. equisimilis H46A (a non-PSGN-associated strain), was predicted to be localized in residues 370 to 374. The epitope domain specific for monoclonal antibody RU-1, which reacts only with the PSGN-associated SKase, was localized to residues 164 to 236.

Amino Acid Sequence↗
PubMed1992

SOS induction in Escherichia coli by infection with mutant filamentous phage that are defective in initiation of complementary-strand DNA synthesis.

We report that the SOS response is induced in Escherichia coli by infection with mutant filamentous phage that are defective in initiation of the complementary (minus)-strand synthesis. One such mutant, R377, which lacks the entire region of the minus-strand origin, failed to synthesize any detectable amount of primer RNA for minus-strand synthesis. In addition, the rate of conversion of parental single-stranded DNA of the mutant to the double-stranded replicative form in infected cells was extremely slow. Upon infection, R377 induced the SOS response in the cell, whereas the wild-type phage did not. The SOS induction was monitored by (i) induction of beta-galactosidase in a strain carrying a dinD::lacZ fusion and (ii) increased levels of RecA protein. In addition, cells infected with R377 formed filaments. Another deletion mutant of the minus-strand origin, M13 delta E101 (M. H. Kim, J. C. Hines, and D. S. Ray, Proc. Natl. Acad. Sci. USA 78:6784-6788, 1981), also induced the SOS response in E. coli. M13Gori101 (D. S. Ray, J. C. Hines, M. H. Kim, R. Imber, and N. Nomura, Gene 18:231-238, 1982), which is a derivative of M13 delta E101 carrying the primase-dependent minus-strand origin of phage G4, did not induce the SOS response. These observations indicate that single-stranded DNA by itself induces the SOS response in vivo.

Coliphages↗
PubMed1992

Correlation of a subset of the pLC plasmids to the physical map of Escherichia coli K-12.

We determined map positions of the Escherichia coli K-12 portions of a subset of the hybrid E. coli-ColE1 plasmids constructed by Clarke and Carbon. The probe DNA of pLC plasmids was labeled with digoxigenine-dUTP, hybridized to the 476 phage clones of the E. coli ordered clone bank miniset, which was adsorbed on a strip of nylon membrane filters, and detected by enzyme-linked immunoassay and a subsequent enzyme-catalyzed color reaction. The total number of Clarke-Carbon plasmids we analyzed was 518, for which chromosomal locations of 297 clones were newly determined in the present study. Another 180 plasmids gave results that agreed with those reported previously, and the remaining 41 plasmids gave map positions different from those described in the previous report. A chromosome map of E. coli which shows the locations of 518 pLC plasmids on it is presented, as well as a table which correlates the pLC plasmids with the clones of the E. coli ordered clone bank miniset on the basis of the hybridization data. We estimate that approximately one-half of the entire genome of E. coli was covered by the pLC plasmids used in this study.

Chromosome Mapping↗