AcpI, a novel isoschizomer of AsuII from Acidiphilium cryptum 25H, recognizes the sequence 5'TT decreases CGAA3'.
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Base Sequence↗
Biomedical subjects
H Kotani
Publications and source records attributed to H Kotani.
Elimination of mycoplasmas from cell cultures by a novel soft agar technique.
Mycoplasmal infection of cell cultures remains a significant threat to diagnostic and research procedures. In certain defined situations, curing of mycoplasmal infected cultures is a reasonable exercise. Four methods of curing were compared: treatment with BM-cycline, 5 bromouracil, use of specific antisera and treatment of infected cells suspended in soft agar with antibiotics. Antisera treatments were of low efficiency of curing: 50%. None of nine infected cell lines treated with 5-bromouracil were consistently cured of mycoplasmas. The use of BM-cycline was effective for some, but not all lines and required long periods of treatment, 12-21 days. 35 naturally or deliberately infected cultures were treated in soft agar a total of 119 times. This procedure which consisted of suspending infected cultures in soft agar containing appropriate antibiotics resulted in successful mycoplasmal elimination 118/119 times. This soft agar technique took 1-3 days. In separate studies, it was shown that certain Mycoplasma fermentans strains were resisted to this and other curing methods. This may be due to their intracellular location. Such strains may be more amenable to antibiotics that penetrate mammalian cells. It is concluded that the soft agar technique is a rapid, efficient and reliable method to eliminate cell culture mycoplasmas.
Agar↗
Denture stomatitis and ABO blood types.
To determine the interrelation between ABO blood types of denture wearers, denture plaque accumulation, and denture stomatitis, 442 denture wearers were studied using a simplified culture method, which is convenient to use in the dental office. The degree of plaque accumulation and the occurrence of denture stomatitis varied depending on the blood type of the patients. Especially in blood group O compared with other types, both denture plaque accumulation and denture stomatitis were found to be higher or more severe. These results suggest that the ABO blood group is one of the etiologic factors of denture stomatitis and that denture wearers of blood group O are more susceptible to denture stomatitis.
ABO Blood-Group System↗
Congenital laminar defect of the upper lumbar spine associated with pars defect. A report of eleven cases.
Lumbar spondylolysis generally is considered to be a fatigue fracture of the pars interarticularis, and no unequivocal case of congenital spondylolysis has been reported. The authors describe 11 cases of unilateral spondylolysis (possibly congenital laminar defect associated with pars defect) of the upper lumbar spine. They have roentgenographic characteristics distinct from conventional spondylolysis of the lumbar spine, including hypoplasia of the spinal accessory process, rotation of the spinous process contralaterally to the spondylolysis, and prominent ipsilateral lamina. The anomaly found in these 11 patients probably is congenital, and its clinical significance is not known.
Adolescent↗
Identification and characterization of proteinase K-resistant proteins in members of the class Mollicutes.
Proteins resistant to proteinase K are rare because of the potency, wide pH optimum, and low peptide bond specificity of this enzyme. Previously, only the prion proteins associated with transmissible spongiform encephalopathies, possibly related proteins in the mollicute Spiroplasma mirum, and proteinase K itself have been reported. We identified a new proteinase K-resistant protein, p40-pr, in two strains of Mycoplasma hyorhinis and in extracts of these organisms. p40-pr's are similar to prion proteins in their resistance to high doses of proteinase K and in the reversal of this resistance by strong denaturing conditions. However, p40-pr's were distinct immunologically, in relative molecular mass, and in their method of extraction. Two immunologically related forms of p40-pr were identified on sodium dodecyl sulfate (SDS) gels and Western immunoblots, a 40-kDa species in boiled samples and a 120-kDa species dissociable by boiling in SDS. Reduction with 2-mercaptoethanol did not affect the mass of p40-pr's or the 120-kDa forms. The development of proteinase K resistance of p40-pr correlated to age-dependent increases in organism protein-lipid ratios. p40-pr-like proteinase K-resistant proteins of 46 to 50 kDa were identified in four of eight additional species of the class Mollicutes but not in S. mirum. However, these mycoplasmal proteins did not react with antibody to the denatured 40-kDa form of M. hyorhinis p40-pr purified by electroelution. The chromatographically purified 46-kDa proteinase K-resistant protein of Mycoplasma orale was an arginine deiminase.
Animals↗
Langerhans cells in the developing mucosal epithelium of mice.
The presence of Langerhans cells in the developing mucosal epithelium of mice was investigated by ATPase histochemistry and by electron microscopy. In the stratified squamous epithelium of the tongue and forestomach, a few ATPase-positive Langerhans cells could be recognized at 1 or 2 weeks after birth. They increased in number by 3 weeks of age, but the density was lower than that in the epidermis. The ATPase-positive cells in the mucosal epithelium had longer (18-22 microns) dendritic processes; each cell thus occupied a large space. The mucosal Langerhans cell with elongated dendritic processes may well be responsible for the surveillance of a wide area in the defense system of the mucous membrane. The cells had a clear cytoplasm among keratinocytes in the basal cell layer and, under the electron microscope, showed characteristics similar to those of the epidermal Langerhans cells; Birbeck granules were also recognizable in the cytoplasm.
Adenosine Triphosphatases↗
[A case of graft-versus-host disease following irradiated fresh blood transfusion].
We reported a case of a fatal graft-versus-host disease (GVHD) which developed in a 65-year-old, male patient which was considered to have been induced by irradiated fresh blood donated by his son after a coronary bypass surgery. Fresh blood was obtained from his relatives, and a 15 Gy irradiation was performed before transfusion. The diagnosis of acute GVHD was made by clinical symptoms and histological examinations of the skin and the bone marrow. He died of sepsis on the 19th post-operative day. The HLA typing of the lymphocytes, revealed that the patient had A 2, A 24, Bw 52, Bw 62, Cw 4, DR 2, and his son had A 24, Bw 52, DR 2. A 24 and Bw 52 were homogeneous making his son histocompatible with one of the patient's haplotype. This might well be attributable to the occurrence of GVHD in this case, meaning that 15 Gy irradiation was not sufficient for the prevention of this disease.
Aged↗
Isolation and identification of restriction endonuclease Asp35HI from Acidiphilium species 35H.
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Base Sequence↗
Sse8387I, a new type-II restriction endonuclease that recognizes the octanucleotide sequence 5'-CCTGCAGG-3'.
A type II restriction endonuclease designated Sse8387I was partially purified from Streptomyces sp. 8387. This enzyme cleaved adenovirus 2 DNA at three sites, lambda phage DNA at five sites, and pUC18 and M13mp18 RF DNA at one site each, but did not cleave the DNAs from pBR322, SV40, or phi X174. Sse8387I recognized the octanucleotide sequence 5'-CCTGCA decreases GG-3', cleaving where shown by the arrow. Sse8387I is the first restriction endonuclease to be reported that recognizes an octanucleotide sequence consisting of all four nucleotides, G, A, T, and C. The frequency of occurrence of Sse8387I sites within sequenced regions of primate genomes was 2.4 times that of NotI sites.
Adenoviruses, Human↗
Cloning, nucleotide sequence, and expression of the HincII restriction-modification system.
Two genes, coding for the HincII from Haemophilus influenzae Rc restriction-modification system, were cloned and expressed in Escherichia coli RR1. Their DNA sequences were determined. The HincII methylase (M.HincII) gene was 1,506 base pairs (bp) long, corresponding to a protein of 502 amino acid residues (Mr = 55,330). The HincII endonuclease (R.HincII) gene was 774 bp long, corresponding to a protein of 258 amino acid residues (Mr = 28,490). The amino acid residues predicted from the R.HincII and the N-terminal amino acid sequence of the enzyme found by analysis were identical. These methylase and endonuclease genes overlapped by 1 bp on the H. influenzae Rc chromosomal DNA. The clone, named E. coli RR1-Hinc, overproduced R.HincII. The R.HincII activity of this clone was 1,000-fold that from H. influenzae Rc. The amino acid sequence of M.HincII was compared with the sequences of four other adenine-specific type II methylases. Important homology was found between tne M.HincII and these other methylases.
Adenine↗
Microbiological cultivation of Mycoplasma hyorhinis from cell cultures.
The failure of many cell culture isolates of Mycoplasma hyorhinis to grow on microbiological media has stressed the need for alternate assays to detect these organisms. The use of freshly prepared yeast extract in mycoplasmal media together with incubation in 5% CO2/air successfully detected M. hyorhinis in 12 of 12 infected cultures. These were not detected by the use of conventional mycoplasmal media using aerobic or anaerobic incubation. This assay may also be helpful in detection of other mycoplasmal species commonly isolated from cell cultures.
Cells, Cultured↗
Mutational analysis of structure--activity relationships in human tumor necrosis factor-alpha.
To determine the region of human tumor necrosis factor-alpha (TNF-alpha), essential for cytotoxic activity against mouse L-M cells, single amino-acid-substituted TNF-alpha mutant proteins (muteins) were produced in Escherichia coli by protein engineering techniques. An expression plasmid for TNF-alpha was mutagenized by passage through an E. coli mutD5 mutator strain and by oligonucleotide-directed mutagenesis. Approximately 100 single amino-acid-substituted TNF-alpha muteins were produced and assayed for cytotoxic activity. The cytotoxic activities of purified TNF-alpha muteins, e.g. TNF-31T, -32Y, -82D, -85H, -115L, -141Y, -144K and -146E, were less than 1% of that of parent TNF-alpha. These results indicate that the integrity of at least four distinct regions of the TNF-alpha molecule is required for full biological activity. These regions are designated as follows: region I, from position 30 to 32; region II, from position 82 to 89; region III, from position 115 to 117; region IV, from position 141 to 146. In addition, TNF-141Y could not completely compete with parent TNF-alpha for binding to the receptor. This demonstrates that region IV, and at least aspartic acid at position 141, must be involved in the TNF receptor binding site.
Amino Acid Sequence↗
Genomic heterogeneity of canine ureaplasmas revealed by restriction endonuclease analysis.
Genomic DNAs of ureaplasma strains isolated from dogs were analyzed by restriction endonucleases. The cleavage patterns revealed by agarose gel electrophoresis, which were characteristic for each strain examined, may provide a direct indication for the genomic index among ureaplasma strains, and can be used as an epidemiological marker in ureaplasma infections.
Animals↗
[Clinical survey on denture stomatitis. 2. The relation between the maintenance of denture and denture stomatitis].
A previous study suggested that denture plaque played an etiologic role in denture stomatitis. In daily practice, it is important to instruct denture wearers about denture plaque control. In this study, the major areas of investigation were the degree of denture stomatitis and denture plaque, denture hygiene habits, wearing habits and age of present denture. 643 patients wearing dentures (1981-1988) were selected, from 47 to 91 years old. The following results were obtained. 1. About 50 percent of the 643 patients used brush for cleaning their dentures. The number of people wearing dentures during sleep was fewer than those who preferred removal of dentures during sleep. 2. Soaking dentures in denture cleanser reduced the degree of denture stomatitis and denture plaque. 3. The patients wearing dentures during sleep showed much more occurrence of denture stomatitis than those who had not that habit. 4. Denture stomatitis occurred frequently with increasing the age of the present denture. The present results indicate that the occurrence of denture stomatitis is closely related to the quality of the maintenance of denture.
Aged↗
Purification and properties of a new restriction endonuclease, MxaI, from Myxococcus xanthus F18E.
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Base Sequence↗
A human IL-1 alpha derivative which lacks prostaglandin E2 inducing activity and inhibits the activity of IL-1 through receptor competition.
In order to segregate multiple activities ascribed to IL-1, various human IL-1 alpha derivatives were produced by recombinant DNA technology. A derivative substituted at the 151st Asp with Tyr(termed to be TN-55) showed unique characteristics. TN-55 lost the PGE2 inducing activity in a human osteosarcoma cell line (MG-63) and growth promoting activity for a human dermal fibroblast cell line (CCD-27Sk), but partially remained LAF activity in mouse thymocyte, cytostatic activity against a human melanoma cell line (A-375) and IL-2 inducing activity in a human T cell line (HSB.2). Although TN-55 bound to the receptor on MG-63 cells with a similar affinity as native IL-1 alpha, TN-55 not only failed in inducing PGE2 production but also antagonized the PGE2 inducing action of IL-1 alpha or IL-1 beta. Thus, TN-55 seems to work as a receptor antagonist. Moreover, TN-55 did not stimulate ACTH secretion in rats in vivo. On the other hand, TN-55 induced PGE2 production in a rabbit dermal fibroblast cell line (RAB-9) and exhibited pyrogenicity in rabbits in vivo. These data suggest that TN-55 has different species-cross reactivity from native IL-1 alpha. In conclusion, multiple biological activities of IL-1 alpha can be segregated by substituting one amino acid. TN-55 may be an ideal IL-1 agonist which lacks inflammatory characteristics of IL-1 (e.g. PGE2-dependent activities) in human but partially retained T lymphocyte stimulating activity and tumor cytostatic activity. In addition, TN-55 may also work as an IL-1 antagonist to block PGE2 production induced by IL-1 through receptor competition.
Adrenocorticotropic Hormone↗
[Effects of ulinastatin on plasma polymorphonuclear leukocyte elastase activity and respiratory function in patients undergoing cardiopulmonary bypass].
Plasma polymorphonuclear leukocyte elastase (PMNE) activity is known to increase during cardiopulmonary bypass (CPB), and is considered to mediate pulmonary tissue damage causing postoperative respiratory failure. We made a clinical study in order to clarify the effects of ulinastatin (Miraclid), a proteolytic enzyme inhibitor, on plasma PMNE activity and respiratory function. Twenty adult patients undergoing coronary artery bypass grafting were divided into 2 groups; 10 to group U and 10 to group C. The patients in group U received 500 U/kg body weight of ulinastatin intravenously before and after CPB, and the patients in group C not receiving the dose served as controls. Arterial blood samples were obtained before the operation, 1 hour after CPB, 3 hours, 1 day and 4 days after the operation. The leukocyte count was significantly lower in group U 1 hour after CPB and 3 hours after the operation compared to group C (p less than 0.01). The plasma PMNE activity rose rapidly after starting CPB and the peak level appeared 1 hour after CPB; 1165 +/- 560 micrograms/l in group U and 1981 +/- 562 micrograms/l in group C (p less than 0.02). There was a significant correlation between the leukocyte reduction and the rate of increase of beta-glucuronidase activity were diminished in group U compared to preoperative values (p less than 0.05). The recovery of the oxygenation index (PaO2/F1O2), which was used for evaluation of the respiratory function, was worse in group C (p less than 0.01). These results suggest that the administration of ulinastatin in patients undergoing CPB is useful for the prevention of the deleterious effects of PMNE on the lung.
Aged↗
[Histology and immunopathology of colonic carcinoma].
The correlation among lymphoid cell infiltration (LI), cell-mediated immunity and immunoglobulin-containing cells distribution in the colon cancer was studied in resected materials of 136 patients with colonic cancer. The results were as follows: 1) LI of the colon cancer tended to be found more commonly in patients with colonic cancer in clinical early stage than advanced stage. 2) The cumulative 5-year survival rate in patients with positive LI was longer than that in the patients with negative or normal LI. 3) In colonic cancer patients with the better prognosis, IgA-containing cells were distributed around the tumor tissues more abundantly than that in the poor ones. 4) These results suggest that the IgA system somewhat participates in cancer immunity.
Colonic Neoplasms↗