ABSTRACT
The aim of this study was to analyze and describe the intrapapillary capillary loops (IPCL), which are a feature of early oral neoplastic lesions, using a narrowband imaging (NBI) system. Forty-one patients (26 men, 15 women; mean age, 52.34 years; range, 23-83 years) presenting with non-neoplastic or neoplastic lesions, and normal cases, were examined using the prototype Evis Lucera Spectrum (Olympus Co.). The images were analyzed and an IPCL classification was devised. All normal cases (n=10) had regularly distributed capillary loops of the same shape (type I). Non-neoplastic lesions (n=8) had mild changes of the capillary loops (types II and III) and neoplastic lesions (n=23) were irregularly distributed and had several loop shapes (types III and IV). The microvascular organization of non-neoplastic lesions was notably different from that of neoplastic lesions. A brownish area was found in five cases of early carcinoma. The narrowband imaging system is a potential approach for clinically analyzing microvascular organization and IPCL. It could be useful for diagnosing oral squamous cell carcinoma at an earlier stage and for determining the margin of resection.
Subject(s)
Capillaries/pathology , Early Detection of Cancer/methods , Endoscopy/methods , Mouth Mucosa/blood supply , Mouth Neoplasms/diagnosis , Adult , Aged , Aged, 80 and over , Carcinoma, Squamous Cell/blood supply , Carcinoma, Squamous Cell/diagnosis , Female , Filtration/instrumentation , Gingiva/blood supply , Gingival Neoplasms/blood supply , Gingival Neoplasms/diagnosis , Humans , Image Processing, Computer-Assisted/methods , Leukoplakia, Oral/blood supply , Leukoplakia, Oral/diagnosis , Lip Diseases/diagnosis , Lip Neoplasms/blood supply , Lip Neoplasms/diagnosis , Male , Middle Aged , Mouth Neoplasms/blood supply , Optical Devices , Stomatitis, Aphthous/diagnosis , Tongue/blood supply , Tongue Neoplasms/blood supply , Tongue Neoplasms/diagnosis , Young AdultABSTRACT
An exclusive measurement has been made of the Coulomb dissociation of the two-neutron halo nucleus 11Li at 70 MeV/nucleon at RIKEN. Strong low-energy (soft) E1 excitation is observed, peaked at about Ex = 0.6 MeV with B(E1) = 1.42(18) e2fm2 for Erel < or = 3 MeV, which was largely missed in previous measurements. This excitation represents the strongest E1 transition ever observed at such low excitation energies. The spectrum is reproduced well by a three-body model with a strong two-neutron correlation, which is further supported by the E1 non-energy-weighted cluster sum rule.
ABSTRACT
Loss of heterozygosity (LOH) on the long arm of chromosome 21 (21q) is observed in several human malignancies. We identified novel tumour suppressor loci on this region in primary oral squamous cell carcinomas (OSCCs). To further determine the role of 21q deletions in oral cavity tumorigenesis, 63 OSCCs were examined for LOH at 21q using 7 microsatellite markers. LOH was observed in 32 of 63 cases (50.8%) that were informative for at least one of the loci analysed. Two distinct deleted regions were identified at chromosomal region 21q11.1. The possible involvement of ANA (abundant in neuroepithelium area), a candidate tumour suppressor gene (TSG) located on 21q11.2--21.1, was also evaluated for 20 OSCCs and 9 OSCC-derived cell lines. 60% of tumours (12/20) and 88.9% (8/9 cell lines) showed absent or reduced mRNA gene expression; only one OSCC case had a nucleotide substitution in the ANA gene. Interestingly, the frequency of the suppressed ANA mRNA expression was greater in stage IV tumours than in earlier stages. In addition, re-expression of the ANA gene mRNA was induced in 4 cell lines after treatment with 5-aza-2'-deoxycytidine, a DNA demethylating agent. These findings demonstrate that there may be at least 2 distinct TSGs on 21q11.1; loss of ANA gene expression could be involved in the progression of human OSCC; and aberrant methylation of the ANA gene promoter may participate in the transcriptional silencing of the gene in oral cancer cells.
Subject(s)
Azacitidine/analogs & derivatives , Chromosomes, Human, Pair 21 , Genetic Predisposition to Disease , Proteins/genetics , Azacitidine/pharmacology , Cell Cycle Proteins , DNA Methylation , Decitabine , Enzyme Inhibitors/pharmacology , Gene Expression Regulation, Neoplastic/drug effects , Gene Silencing , Humans , Loss of Heterozygosity , Mutation , Polymerase Chain Reaction , Polymorphism, Single-Stranded Conformational , RNA, Messenger/genetics , Tumor Cells, CulturedABSTRACT
Frequent allelic imbalances including loss of heterozygosity (LOH) and microsatellite instability (MSI) on the short arm of chromosome 3 (3p) have been found in several types of human cancer. This study was designed to identify the tumor suppressor locus (or loci) on 3p associated with tongue squamous cell carcinoma (SCC). Among 16 patients with tongue SCC tested, 7 (44%) of 16 informative cases showed LOH at one or more loci. Deletion mapping of these 16 tumors revealed two discrete, commonly deleted regions on the chromosome arm. Our data support the notion that tumor suppressor gene(s) contributing to the progression of tongue squamous cell carcinoma reside on 3p24 and 3p25.
Subject(s)
Carcinoma, Squamous Cell/genetics , Chromosomes, Human, Pair 3 , Gene Deletion , Genes, Tumor Suppressor , Tongue Neoplasms/genetics , Chi-Square Distribution , DNA, Neoplasm/analysis , Female , Humans , Loss of Heterozygosity , Male , Microsatellite Repeats , Polymerase Chain ReactionABSTRACT
The p16/CDKN2 (cyclin dependent kinase number 2) gene is known to be one of the negative regulators of the cell cycle. Aberrant 5'CpG island methylation is one of the most important mechanisms of p16/CDKN2 gene promoter region alteration. We studied 8 oral squamous cell carcinoma cell lines and 25 primary tumor tissues for the p16/CDKN2 gene and its expression by PCR-SSCP, MSP, RT-PCR, and immunohistochemical methods to determine the mechanism and the potential biological significance of p16/CDKN2 gene inactivation. In primary tumors, no p16/CDKN2 gene mutations were found by PCR-SSCP. However, hypermethylation of the CpG sites of p16/CDKN2 gene was observed in 48% (12/25) cases of primary tumors and in 50% (4/8) of cell lines. To verify the p16 mRNA expression, we employed RT-PCR and observed decreased or lacked p16 mRNA in 44% (11/25) of primary tumor tissues. In addition, hypermethylation was observed in 6 of the above 11 cases (55%). An immunohistochemistry assay was also performed with the primary tumor tissues, and a semi-quantitative method was used to evaluate the staining intensity of p16 protein. We observed 52% (13/25) negative nuclear staining. When we compared these results with clinicopathological stages, there was no statistical significance. These findings suggest that hypermethylation of p16/CDKN2 promoter region may be associated with p16/CDKN2 gene alteration.
Subject(s)
Carcinoma, Squamous Cell/genetics , Carcinoma, Squamous Cell/metabolism , Cyclin-Dependent Kinase Inhibitor p16/biosynthesis , Mouth Neoplasms/genetics , Mouth Neoplasms/metabolism , Aged , Aged, 80 and over , Chi-Square Distribution , Cyclin-Dependent Kinase Inhibitor p16/analysis , Cyclin-Dependent Kinase Inhibitor p16/genetics , DNA Methylation , DNA, Neoplasm/analysis , DNA, Neoplasm/metabolism , Female , Genes, p16 , Humans , Immunoenzyme Techniques , Loss of Heterozygosity , Male , Middle Aged , Polymorphism, Single-Stranded Conformational , Reverse Transcriptase Polymerase Chain Reaction , Tumor Cells, CulturedABSTRACT
AIM: To analyse the capacity for epithelial differentiation in synovial sarcoma using a new human cell line. METHODS: A new human cell line, KU-SS-1, was established from a monophasic, spindle cell type of synovial sarcoma by grafting those cells on to severe combined immunodeficient (SCID) mice and then transferring them to in vitro culture systems. The KU-SS-1 cells were characterised by light and electron microscopy, and by immunohistochemical, flow cytometric, and cytogenetic analysis. RESULTS: Primary tumour and cultured cells at passage 20 showed a positive reaction for vimentin, which is a mesenchymal marker. After 40 passages, subcultured cells were injected into SCID mice to induce further tumours. These advanced subcultured cells and the tumour cells that they induced were positive for cytokeratin, an epithelial marker, and exhibited epithelial ultrastructural features such as intermediate junctions. Furthermore, two colour immunofluorescent analysis for proliferating nuclear cell antigen (PCNA) and intermediate filaments showed that a large number of PCNA expressing cells were positive for vimentin, and that part of this fraction also expressed cytokeratin. The existence of cells with reactivity for these three markers indicated that, in this cell line, a fraction with high proliferating capacity had both mesenchymal and epithelial markers. In addition, cytogenetically, this cell line expressed the SYT-SSX chimaeric transcript as a result of the t(X;18) (p11;q11) translocation. CONCLUSIONS: A human synovial sarcoma cell line was established and stably maintained in cell culture for more than 70 passages. In addition, this cell line showed epithelial differentiation, which supports the hypothesis that synovial sarcoma is a carcinosarcoma like tumour with true epithelial differentiation. This cell line will be a useful tool for investigating the nature of this tumour and will contribute to clinical studies.
Subject(s)
Cell Transformation, Neoplastic/pathology , Knee Joint , Sarcoma, Synovial/pathology , Tumor Cells, Cultured/pathology , Adult , Animals , Epithelial Cells/pathology , Female , Flow Cytometry , Humans , Karyotyping , Mice , Mice, SCID , Neoplasm Transplantation , Proliferating Cell Nuclear Antigen/metabolism , Reverse Transcriptase Polymerase Chain Reaction/methods , Sarcoma, Synovial/genetics , Transplantation, HeterologousABSTRACT
The action of 5-hydroxytryptamine (5-HT) via the 5-HT1A receptor on dissociated rat dorsal raphe neurons was characterized under the whole-cell mode by using the nystatin-perforated patch-clamp technique. Under voltage-clamp conditions, 5-HT induced an inwardly rectifying K+ current (I5-HT) in a concentration-dependent manner. I5-HT was mimicked by 8-OH-DPAT and buspirone, which are both 5-HT1A receptor agonists. I5-HT was reversibly blocked by such 5-HT1A receptor antagonists as (S)-UH-301 a 5-HT4 receptor antagonist. I5-HT was antagonized concentration-dependently by such K+ channel blockers as quinine, Ba2+ and 4-aminopyridine but was relatively insensitive to both CS+ and tetraethylammonium. When the neurons were loaded with guanosine 5'-O-3-thiotriphosphate through a patch pipette, the K+ current induced by 5-HT became irreversible. N-ethylmaleimide (NEM), a sulfhydryl alkylating agent, irreversibly blocked I5-HT. The intracellular perfusion with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA), a Ca2+ chelator, or neomycine, a phospholipase C inhibitor, never significantly affected the 5-HT-induced response. 12-Myristate 13-acetate diester (PMA), a protein kinase C (PKC) activator, had only a weak inhibitory effect on I5-HT, and staurosporine, a PKC inhibitor, failed to significantly occlude I5-HT. Therefore, the K+ conductance activated via the 5-HT1a receptor of dorsal raphe neurons was thus characterized by the sensitivity to such K+ channel blockers as quinine, Ba2+ and 4-aminopyridine. Moreover, G protein which is NEM-sensitive and can couple to the 5-HT1A receptor, is thus considered to activate the inwardly rectifying K+ conductance without being mediated by such second messengers as Ca2+ and PKC.
Subject(s)
Neurons/physiology , Potassium Channels/metabolism , Raphe Nuclei/physiology , Receptors, Serotonin/physiology , Animals , Calcium/physiology , Electrophysiology , Enzyme Inhibitors/pharmacology , Ethylmaleimide/pharmacology , Guanine Nucleotides/pharmacology , In Vitro Techniques , Membrane Potentials/drug effects , Membrane Potentials/physiology , Patch-Clamp Techniques , Potassium Channels/drug effects , Protein Kinase C/antagonists & inhibitors , Protein Kinase C/physiology , Raphe Nuclei/cytology , Rats , Rats, Wistar , Serotonin/pharmacology , Serotonin Antagonists/pharmacology , Serotonin Receptor Agonists/pharmacologyABSTRACT
The augmentative effect of liposomes containing adriamycin (LADM) in the host-tumour immune mechanism was assessed using 9,10-dimethyl-1,2-benzanthracene induced rat malignant fibrous histiocytoma (MFH). The antitumour effect of LADM was analyzed using a double grafted tumour system in which F344 rats first received simultaneous s.c. inoculations of MFH cells in both right and left flanks and were then injected with 0.2 mg of LADM into the right tumour on Day 10, 12, and 14. The growth of a remote, non-treated tumour was strongly inhibited, and the infiltration of CD8+ or CD4+ cells at the tumour periphery was histologically revealed. This inhibition was not observed in F344 nu/nu athymic rats. Winn neutralizing assay with T cell-rich splenic lymphocytes from each drug-treated MFH-bearing rat showed that complete regression of the tumour at a low effector/target ratio occurred only in the LADM-treated group. Furthermore, the tumouricidal effects were demonstrated in coexistence with CD8+ cells and CD4+ cells by assay with FACS sorting splenic lymphocytes. These results strongly suggest that intratumoural administration of LADM caused the systemic augmentation of the MFH-bearing host immune mechanism, and that the augmented response after LADM treatment was induced by the cytotoxic CD8+ lymphocytes dependent on the CD4+ lymphocytes.
Subject(s)
Antibiotics, Antineoplastic/therapeutic use , Doxorubicin/therapeutic use , Histiocytoma, Benign Fibrous/therapy , Immunotherapy, Adoptive , T-Lymphocytes/immunology , 9,10-Dimethyl-1,2-benzanthracene , Animals , Antibiotics, Antineoplastic/administration & dosage , Cell Division , Doxorubicin/administration & dosage , Drug Carriers , Histiocytoma, Benign Fibrous/chemically induced , Histiocytoma, Benign Fibrous/immunology , Histiocytoma, Benign Fibrous/pathology , Liposomes , Lymphocyte Activation/drug effects , Lymphocytes, Tumor-Infiltrating/pathology , Male , Rats , Rats, Inbred F344 , Rats, Nude , Spleen/immunology , T-Lymphocytes/drug effectsABSTRACT
Electrical and pharmacological properties of currents induced by compounds having affinities for putative sigma receptors were investigated with NCB20 cells by use of the whole-cell patch-clamp technique. Antipsychotics and naloxone induced inward currents with a decrease in membrane conductance at a holding potential of -60 mV. The rank order of potency for compounds inducing these currents was bromperidol > haloperidol > mosapramine = clocapramine > carpipramine > chlorpromazine > remoxipride > naloxone. Sulpiride, which does not have affinity for sigma receptors, induced inward currents only slightly. Haloperidol-induced currents were not affected by the pretreatments with 10 microM of sulpiride, dopamine, atropine, N-methyl-D-aspartate, 2-amino-7-phosphonoheptanoic acid, morphine or A23187, 100 nM of ICS 205-930, 100 microM of forskolin, 1 microM of phorbol-12,13-dibutyrate, or 100 ng/ml of cholera or pertussis toxins. The reversal potential of the currents induced by haloperidol, naloxone or remoxipride was dependent on the concentration of external or internal potassium. These results indicate that the currents induced by the tested compounds are due to blockade of tonic, outward potassium currents and suggest that these agents act on putative sigma receptors and that the second messenger systems within the cell are not essential for the coupling between the receptors and the channels.
Subject(s)
Brain Neoplasms/metabolism , Neuroblastoma/metabolism , Potassium Channels/metabolism , Receptors, sigma/drug effects , Animals , Antipsychotic Agents/pharmacology , Cricetinae , Electrophysiology , Haloperidol/pharmacology , Ligands , Membrane Potentials/drug effects , Mice , Naloxone/pharmacology , Potassium Channels/drug effects , Remoxipride/pharmacology , Tumor Cells, CulturedABSTRACT
1. The effects of benzodiazepine receptor (BZR) partial agonists, Y-23684 and CL218,872, were compared with its full agonist, diazepam, on gamma-aminobutyric acid (GABA)-induced Cl- current (ICl) in acutely dissociated rat cerebral cortex (CTX), cerebellar Purkinje (CPJ) and spinal ventral horn (SVH) neurones, by the whole-cell mode patch-clamp technique. 2. The GABA-induced responses were essentially the same in both SVH and CPJ neurones, but the KD value of the GABA response in CTX neurone was lower than those in the other two brain regions. 3. Enhancement of the GABA response by the two partial agonists was about one-third of that by diazepam in the SVH neurones (where type II subtype of BZR, BZ2, is predominant), whereas these partial agonists potentiated the GABA response as much as diazepam in CPJ neurones (where the type I subtype of BZR, BZ1, is predominant). In CTX neurones where both type I and II variants are expressed, the augmentation ratio of the GABA response by diazepam was between the values in CPJ and SVH neurones. 4. In concentration-response relationships of BZR partial agonists, the threshold concentrations, KD values and maximal augmentation ratio of the GABA response were similar in all CTX, CPJ and SVH neurones. Also, in all preparations, the threshold concentration and KD values of diazepam action were 10 fold less than those induced by partial agonists. 5. All BZR agonists shifted the concentration-response relationship for GABA to the left without changing the maximum current amplitude, indicating that activation of both BZ1 and BZ2 increase the affinity of the GABAA receptor for GABA. 6. The results are important in clarifying the mechanism of anxiety and might explain the anxioselectivity of BZR partial agonists.
Subject(s)
Central Nervous System/cytology , Neurons/drug effects , Receptors, GABA-A/drug effects , Aging/metabolism , Animals , Benzothiepins/pharmacology , Central Nervous System/drug effects , Cerebral Cortex/cytology , Cerebral Cortex/drug effects , Cerebral Cortex/metabolism , Diazepam/pharmacology , In Vitro Techniques , Membrane Potentials/drug effects , Purkinje Cells/drug effects , Purkinje Cells/metabolism , Pyridazines/pharmacology , Rats , Rats, Wistar , Spinal Cord/cytology , Spinal Cord/drug effects , Spinal Cord/metabolism , Tranquilizing Agents/pharmacology , gamma-Aminobutyric Acid/pharmacologyABSTRACT
Carbohydrate antigens of the serotype c/Lancefield group C "Streptococcus milleri" were extracted by autoclaving whole cells of the type c reference strain K51Y. The type c and group C antigen molecules were separated and partially purified by a DEAE-Sephadex A-25 column chromatography followed by Sephadex G-100 gel filtration. The purified type c antigen and group C antigen were homogeneous in the double diffusion and in the immunoelectrophoresis. The type c antigen was composed principally of glycerol, rhamnose, glucose and N-acetylglucosamine in a molar ratio of 0.22:0.27:1.00:0.48. The quantitative precipitin inhibition test indicated that N-acetylglucosamine played a major role in immunodeterminant structure. Thus, the type c antigen of "S. milleri" is a new carbohydrate type antigen and is immunochemically different from the Ottens-type antigen III found occasionally in group C streptococci. In contrast, the group C antigen preparation contained a high proportion of N-acetylgalactosamine in addition to glycerol, rhamnose, glucose and N-acetylglucosamine, and the N-acetylgalactosamine residue was involved in the immunodominant epitope, being in good agreement with the previously proposed chemical structure of the group antigen. N-acetylgalactosamine was detected in the autoclaved extracts of a nontypeable/group C strain but not of a type c/ungroupable strain.
Subject(s)
Antigens, Bacterial/chemistry , Streptococcus/immunology , Acetylglucosamine/analysis , Antigens, Surface/chemistry , Carbohydrates/analysis , Carbohydrates/immunology , Immunodiffusion , Streptococcus/classificationABSTRACT
A method was introduced for the estimation of total dietary fiber (TDF) intake of a population using a menu-oriented questionnaire and a menu-based calculation table. TDF intake correlated well with age in a population investigated, and in younger generations TDF consumption was very low (less than 11.5 g/day in teenagers). The similar results were obtained from the calculation using data of National Nutrition Survey (10.7 g/day). The foodstuffs they consumed were more processed and refined. This fact suggested that in younger generations a future resumption of their present eating habits might produce a serious lack of TDF intake in later years. To clarify the optimal level of TDF intake for the upper limit of recommended daily allowance (RDA) of an average Japanese, the following were measured and calculated. (I) Estimation of recent TDF intake (1990) and of 30 and 50 years ago (1955, 1935), based on TDF data of foodstuffs by the enzymatic-gravimetric method. (II) Measurement of the TDF of model duplicate meals and model composite diets for the average Japanese in 1985 using the same assay method. (III) Conversion of a recommendation of 20-35 g/day for American into RDA for Japanese considering energy consumption and lower fat intake. (IV) Re-estimation of the literature data on the adverse effects of DF on the human mineral balance considering the insufficient calcium intake of Japanese. The results indicated an RDA of 10-12 g TDF/1,000 kcal fit better for an average Japanese.
Subject(s)
Dietary Fiber/analysis , Nutritional Requirements , Adolescent , Adult , Age Factors , Aged , Female , Humans , Japan , Male , Middle Aged , Nutrition SurveysABSTRACT
A series of novel 7-substituted 1-cyclopropyl-6,8-difluoro-1, 4-dihydro-4-oxo-3-quinolinecarboxylic acids have been prepared and tested for antibacterial activities and for convulsive activities in combination with nonsteroidal antiinflammatory drug. Structure-activity relationships revealed that 7-(2-(aminomethyl)morpholino) derivative 28 had a better Gram-positive activity than the reference quinolones, such as ciprofloxacin, norfloxacin, and ofloxacin. Its Gram-negative activity was equipotent with those of norfloxacin and ofloxacin but was inferior to that of ciprofloxacin. In mouse systemic infection models, 28 showed an excellent therapeutic efficacy which might result from the potent antibacterial activity and suitable physicochemical properties. Convulsive activities of 7-morpholino derivatives in combination with nonsteroidal antiinflammatory drug fenbufen or its metabolite biphenylacetic acid markedly diminished as compared to those of 7-piperazino derivatives in the electrophysiological, biochemical, and behavioral experiments. These results suggest that 28 (Y-26611) is a novel quinolone with reduced neurotoxic excitatory adverse reaction.
Subject(s)
4-Quinolones , Anti-Infective Agents/chemical synthesis , Morpholines/chemical synthesis , Animals , Anti-Infective Agents/chemistry , Anti-Infective Agents/therapeutic use , Escherichia coli/drug effects , Male , Mice , Mice, Inbred ICR , Microbial Sensitivity Tests , Morpholines/chemistry , Morpholines/therapeutic use , Quinolones/chemical synthesis , Quinolones/chemistry , Quinolones/therapeutic use , Staphylococcus aureus/drug effects , Structure-Activity RelationshipABSTRACT
Of the 29 'Streptococcus milleri' strains tested, all thirteen Streptococcus intermedius (DNA homology group 2) strains but none of the thirteen Streptococcus anginosus (group 1) strains produced beta-N-acetylglucosaminidase, beta-N-acetylgalactosaminidase, alpha-N-acetylneuraminidase, beta-galactosidase, alpha-glucosidase, and hyaluronidase. The three Streptococcus constellatus (group 3) strains produced only the latter two. Glycosidase production divided 274 clinical isolates into 103 S. anginosus, 101 S. intermedius, and 70 S. constellatus strains. Generally, strains of S. anginosus and S. intermedius were non-beta-haemolytic. API II and biotype Ia (lactose positive), but the former contained almost all API III strains and belonged to Lancefield group A/serotype a (A/a), -/b, C/c, -/d, -/e, F/f or G/k, and the latter included most of biotype IId (lactose negative) and serovar -/g, -/h, -/i or -/j. S constellatus strains were beta-, alpha- or gamma-haemolytic, of API I or II but mostly biotype Ib (lactose negative), and of F/- or -/b. S. intermedius was a major member of the oral isolates. Non-oral isolates were virtually all S. anginosus (mainly urogenital isolates) or S. constellatus (the other systemic isolates).
Subject(s)
Streptococcus/classification , Adult , Bacterial Typing Techniques , Carbohydrate Metabolism , Child , Female , Glycoside Hydrolases/biosynthesis , Hemolysis , Humans , Infant, Newborn , Mouth/microbiology , Serotyping , Species Specificity , Streptococcal Infections/microbiology , Streptococcus/enzymology , Streptococcus/metabolism , Urogenital System/microbiologyABSTRACT
Of a total 148 strains of Streptococcus milleri group, 66 agglutinated sheep erythrocytes. The haemagglutinating strains were confined to serotypes g, h, i, j or were untypeable, and had no Lancefield group antigens (A to G and K), all of which have been shown to belong to Streptococcus intermedius. Cell surface hydrophobicity did not significantly differ even between the agglutinating and non-agglutinating S. intermedius strains. The haemagglutinating activity of strain 0813-1 (serotype i) was partially sensitive to heat (100 degrees C, 30 min) or trypsin (1 mg/ml, 30 min) treatment, but completely lost after the heat and subsequent trypsin treatments. Only L-arginine, L-lysine (100 mM), mucin, and fetuin (1 mg/ml) partially inhibited the haemagglutination with native bacterial cells but completely inhibited that with heated cells, whereas none was inhibitory in the reaction with trypsin-treated cells. The results suggest that at least two haemagglutinins are involved in the agglutination of the S. intermedius strain and that the heat-stable but trypsin-sensitive haemagglutinin recognizes the receptors on the erythrocyte surface which contain L-arginine and L-lysine at the reactive site.
Subject(s)
Erythrocytes/physiology , Hemagglutination/physiology , Hot Temperature , Streptococcus/physiology , Trypsin/pharmacology , Amino Acids/pharmacology , Animals , Glycoproteins/pharmacology , Hemagglutination/drug effects , Humans , Species SpecificityABSTRACT
Strains of oral Streptococcus milleri group were compared with the type strains of Streptococcus anginosus, S. intermedius, and S. constellatus by DNA-DNA hybridization at 60 degrees C. Of the 29 strains tested, twelve, twelve, and two strains were closely related to S. anginosus ATCC 33397T, S. intermedius ATCC 27335T, and S. constellatus ATCC 27823T, respectively. Generally, the strains classified in the S. anginosus group in DNA homology were non-beta-haemolytic and belonged to API S. milleri II and biotype I (lactose fermenting), whereas the strains of S. constellatus group were beta-haemolytic and belonged to API S. milleri I and biotype II (lactose non-fermenting). The strains of the S. intermedius group were all non-beta-haemolytic and belonged to API S. milleri II and mostly biotype II (lactose fermenting). Furthermore, all except one strain of the S. anginosus group were Lancefield group A/serotype a (A/a), ungroupable/serotype b (-/b), C/c, -/d, -/e, F/f or G/k, whereas those of the S. constellatus group were F/-. The strains of the S. intermedius group were -/g, -/h, -/i, -/j, or -/-.
Subject(s)
DNA, Bacterial/isolation & purification , Mouth/microbiology , Streptococcus/genetics , DNA Probes , DNA, Bacterial/genetics , Humans , Phenotype , Sequence Homology, Nucleic Acid , Serotyping , Species Specificity , Streptococcus/classification , Streptococcus/isolation & purificationABSTRACT
1. The effect of Y-25130, ((+-)-N-(1-azabicyclo[2.2.2]oct-3-yl)-6-chloro-4-methyl-3-oxo-3,4-dih ydr o- 2H-1,4-benzoxazine-8-carboxamide hydrochloride), a high affinity 5-hydroxytryptamine3 (5-HT3) receptor ligand, was examined on the 5-HT-induced response in dissociated frog dorsal root ganglion (DRG) neurones by use of the extremely rapid concentration-jump ('concentration-clamp') and the conventional whole-cell patch-clamp techniques. 2. 5-HT induced a rapid transient inward current associated with an increase in membrane conductance at a holding potential of -70 mV. The current amplitude increased sigmoidally as 5-HT concentration increased. The half-maximum value (Ka) and the Hill coefficient estimated from the concentration-response curve were 1.7 x 10(-5) M and 1.7, respectively. 3. The current-voltage (I-V) relationship of 5-HT-induced current (I5-HT) showed inward rectification at potentials more positive than -40 mV. The reversal potential (E5-HT) was -11 mV. The E5-HT value was unaffected by total replacement of intracellular K+ by Cs+, indicating that the 5-HT-gated channels might be large cation channels. 4. Both the activation and inactivation phases of I5-HT were single exponentials. The time constants of activation and inactivation (tau a and tau i) decreased with increasing 5-HT concentration. 5. The 5-HT response was mimicked by a selective 5-HT3 receptor agonist, 2-methyl-5-HT, but the maximum response induced was approximately 25% that of 5-HT. The 5-HT response was reversibly antagonized by the 5-HT3 receptor antagonists, ICS 205-930, metoclopramide and Y-25130, but not by a 5-HTIA receptor antagonist, spiperone, and a 5-HT2 receptor antagonist, ketanserin. The half-inhibition concentrations (IC50) were 4.9 x 10-10 M for Y-25130, 4.8 x 10-10 M for ICS 205-930 and 8.6 x 10-9 M for metoclopramide.6. Y-25130 (5 x 10-10 M) caused a rightward shift of the concentration-response curve for 5-HT while decreasing the maximum response.7. The results suggest that Y-25130 is a potent antagonist of the 5-HT3 receptor-channel complex.
Subject(s)
Bridged Bicyclo Compounds, Heterocyclic , Bridged Bicyclo Compounds/pharmacology , Neurons, Afferent/physiology , Oxazines/pharmacology , Serotonin Antagonists , Animals , Ganglia, Spinal/cytology , Ganglia, Spinal/drug effects , In Vitro Techniques , Kinetics , Membrane Potentials/drug effects , Neurons/drug effects , Neurons, Afferent/drug effects , Potassium/pharmacology , Rana catesbeiana , Serotonin/pharmacologyABSTRACT
The distribution of oral Streptococcus milleri carbohydrate type antigens in other viridans streptococcus species was examined. Rantz-Randall extracts of cells of the test strains grown in broth containing glucose were allowed to react with typing or grouping antisera for S. milleri serotypes a-k, or Lancefield groups A-G and K. Of 93 strains comprising more than 12 streptococcal species that included S. mutans and S. sanguis complexes, only 15 S. salivarius strains and one S. mitis strain were immunologically related to S. milleri serotype f. Unlike S. milleri strains, S. salivarius type f strains belonged to Lancefield group K, whereas the S. mitis strain was closely related to S. milleri serotype f but did not react with any of the Lancefield grouping antisera tested. Results suggest that oral S. milleri strains can be distinguished serologically from other oral viridans streptococci and that the typing antisera used in our researches might differentiate S. milleri isolates from the mouth from those associated with systemic purulent infections.
Subject(s)
Antigens, Bacterial/analysis , Carbohydrates/analysis , Mouth Mucosa/microbiology , Streptococcus/immunology , Carbohydrates/immunology , Cross Reactions , Humans , Immunodiffusion , Serotyping , Streptococcus/classificationABSTRACT
The aim of this study was to elucidate the vasodilator mechanisms of barbiturates. In helical strips of dog mesenteric artery, the effects of pretreatment with thiopentone and pentobarbitone on the contractions induced by KCl (2.0 x 10(-2) M) and norepinephrine (10(-5) M) in normal bathing fluid, and those induced by norepinephrine and caffeine (2.5 x 10(-2) M) in Ca(++)-free fluid were tested, and their effects on caffeine-induced contractions in skinned strips were also examined. Thiopentone, at concentrations over 10(-4) M, inhibited the KCl-induced contractions in normal bathing fluid and those induced by caffeine in Ca(++)-free fluid and, at concentrations over 3 x 10(-4) M, inhibited norepinephrine-induced contractions in normal and Ca(++)-free bathing fluids significantly. Pentobarbitone, at concentrations over 3 x 10(-4) M, inhibited KCl- and norepinephrine-induced contractions in normal bathing fluid significantly, whereas contractions in Ca(++)-free fluid induced by norepinephrine and caffeine were inhibited only by a high concentration (10(-3) M) of pentobarbitone. Caffeine-induced contractions of chemically skinned fibres were more susceptible to inhibition by thiopentone than by pentobarbitone. These results suggest that the vasodilator effect of thiopentone is mediated via its intracellular inhibitory effect and that, in contrast, the vasodilator effect of pentobarbitone can be attributed mainly to its Ca(++)-channel blocking action.