ABSTRACT
In this study, we report a tumor that resembled previously reported uncommon tumors histologically similar to ameloblastoma or adenomatoid odontogenic tumor (AOT), showing the formation of hard tissue. We evaluated the histological characteristics by reviewing the literature. The patient was a 19-year old male. The lesion was located from the canine to third molar in the right mandible and was unicystic with a comparatively clear demarcation. The tumor tissue was cystic overall, showing multiple formation of small and large cysts. The tumor tissue resembled a variant form of plexiform ameloblastoma. Formation of dentin and dentinoid was observed in the tumor stroma, whereas formation of enamel was not observed. Very few cases of a variant form of ameloblastoma that shows formation of dentinoid have been reported, and the histological picture in this study closely resembled previously reported "adenoid ameloblastoma with dentinoid".
Subject(s)
Ameloblastoma/pathology , Dentin/pathology , Mandibular Neoplasms/pathology , Adult , Coloring Agents , Cuspid/pathology , Dental Arch/pathology , Follow-Up Studies , Humans , Male , Molar, Third/pathology , Neoplasm Recurrence, Local/pathologyABSTRACT
BACKGROUND: Influenza virus RNA polymerase is a multifunctional enzyme that catalyses both transcription and replication of the RNA genome. The function of the influenza virus RNA polymerase PA subunit in viral replication is poorly understood, although the enzyme is known to be required for cRNA --> vRNA synthesis. The protease related activity of PA has been discussed ever since protease-inducing activity was demonstrated in transfection experiments. RESULTS: PA protein was highly purified from insect cells infected with the recombinant baculovirus carrying PA cDNA, and a novel chymotrypsin-type serine protease activity was identified with the synthetic peptide, Suc-LLVY-MCA, in the PA protein. [3H]DFP was crosslinked with PA and a mutational analysis revealed that serine624 was as an active site for the protease activity. CONCLUSIONS: These results constitute the demonstration of protease activity in PA subunit of the influenza virus RNA polymerase complexes.
Subject(s)
DNA-Directed RNA Polymerases/metabolism , Orthomyxoviridae/enzymology , Serine Endopeptidases/metabolism , Serine/metabolism , Amino Acid Sequence , Animals , Base Sequence , Binding Sites , Catalysis , Chromatography, Liquid , DNA Primers , DNA-Directed RNA Polymerases/chemistry , DNA-Directed RNA Polymerases/genetics , DNA-Directed RNA Polymerases/isolation & purification , Electrophoresis, Polyacrylamide Gel , Hydrogen-Ion Concentration , Hydrolysis , Recombinant Proteins/chemistry , Recombinant Proteins/genetics , Recombinant Proteins/isolation & purification , Recombinant Proteins/metabolism , Serine Endopeptidases/chemistry , Serine Endopeptidases/genetics , Serine Endopeptidases/isolation & purification , SpodopteraABSTRACT
A novel potentiator of nerve growth factor (NGF), NG-061, which had been isolated from the fermentation broth of Penicillium minioluteum F-4627, was synthesized from methoxybenzoquinone and phenylacetylhydrazine in a single step. A series of acyl hydrazone derivatives were also synthesized and their potentiator activity of neurotrophic effect of NGF on neurite outgrowth was evaluated by assay with a rat pheochromocytoma cell line PC12.
Subject(s)
Hydrazines/chemical synthesis , Nerve Growth Factor/agonists , Phenylacetates/chemical synthesis , Animals , Drug Evaluation, Preclinical , Hydrazines/chemistry , Hydrazines/pharmacology , Molecular Structure , PC12 Cells , Phenylacetates/chemistry , Phenylacetates/pharmacology , Rats , Spectrum AnalysisABSTRACT
FD-891 belongs to a group of 18-membered macrolides, and is a structural analogue of a specific inhibitor of vacuolar type H+-ATPase, concanamycin A (CMA). In our previous work, we have shown that CMA specifically inhibits perforin-dependent cytotoxic T lymphocyte (CTL)-mediated cytotoxicity through the degradation and inactivation of perforin, although CMA does not affect Fas ligand (FasL)-dependent cytotoxicity. Here, we show that FD-891 potently prevents not only perforin-dependent but also FasL-dependent CTL-mediated killing pathways by blocking CTL-target conjugate formation. In contrast to CMA, FD-891 was unable to inhibit vacuolar acidification and only slightly decreased the perforin activity in lytic granules. FD-891 blocked granule exocytosis in response to anti-CD3, mainly owing to the lack of CTL binding to immobilized anti-CD3. The conjugate formation was markedly inhibited only when effector cells were pretreated with FD-891. Consistent with these observations, fluorescence-activated cell sorter (FACS) analysis for cell surface receptors revealed that FD-891 significantly reduced the expression of the T-cell receptor (TCR)/CD3 complex. These data suggest that the blockage of conjugate formation and subsequent target cell killing might be at least partly owing to FD-891-induced down-regulation of the TCR/CD3 complex.
Subject(s)
Anti-Bacterial Agents/pharmacology , Cytotoxicity, Immunologic/drug effects , Macrolides , T-Lymphocytes, Cytotoxic/drug effects , Animals , CD3 Complex/metabolism , Exocytosis/drug effects , Fas Ligand Protein , Hydrogen-Ion Concentration/drug effects , Ligands , Membrane Glycoproteins/antagonists & inhibitors , Membrane Glycoproteins/immunology , Membrane Glycoproteins/metabolism , Mice , Mice, Inbred BALB C , Perforin , Pore Forming Cytotoxic Proteins , Receptors, Antigen, T-Cell/metabolism , T-Lymphocytes, Cytotoxic/immunology , Vacuoles/drug effects , Vacuoles/metabolism , fas Receptor/immunology , fas Receptor/metabolismSubject(s)
Antineoplastic Agents/pharmacology , Cell Survival/drug effects , Fungi/chemistry , Furans/pharmacology , Antineoplastic Agents/chemistry , Antineoplastic Agents/isolation & purification , Furans/chemistry , Furans/isolation & purification , HL-60 Cells , Humans , Magnetic Resonance Spectroscopy , Mass SpectrometryABSTRACT
This paper focuses on the inhibitory mechanism of E-64 and its derivatives (epoxysuccinyl-based inhibitors) with some cysteine proteases, based on the binding modes observed in the x-ray crystal structures of their enzyme-inhibitor complexes. E-64 is a potent irreversible inhibitor against general cysteine proteases, and its binding modes with papain, actinidin, cathepsin L, and cathepsin K have been reviewed at the atomic level. E-64 interacts with the Sn subsites of cysteine proteases. Although the Sn-Pn (n = 1-3) interactions of the inhibitor with the main chains of the active site residues are similar in respective complexes, the significant difference is observed in the side-chain interactions of S2-P2 and S3-P3 pairs because of different residues constituting the respective subsites. E-64-c and CA074 are representative derivatives developed from E-64 as a clinical usable and a cathepsin B-specific inhibitors, respectively. In contrast with similar binding/inhibitory modes of E-64-c and E-64 for cysteine proteases, the inhibitory mechanism of cathepsin B-specific CA074 results from the binding to the Sn' subsite.
Subject(s)
Cysteine Endopeptidases/chemistry , Cysteine Proteinase Inhibitors/chemistry , Cysteine Proteinase Inhibitors/pharmacology , Leucine/analogs & derivatives , Binding Sites , Crystallography, X-Ray , Cysteine Endopeptidases/metabolism , Cysteine Proteinase Inhibitors/chemical synthesis , Drug Design , Epoxy Compounds , Leucine/chemical synthesis , Leucine/chemistry , Leucine/pharmacology , Models, Molecular , Protein Conformation , Static Electricity , Structure-Activity Relationship , Succinic AcidABSTRACT
The structure of NG-061, a new potentiator of nerve growth factor (NGF) isolated from Penicillium minioluteum F-4627, was determined by spectroscopic analysis and X-ray diffraction method to be phenylacetic acid 2-(2-methoxy-4-oxocyclohexa-2,4-dienylidene)-hydrazide.
Subject(s)
Hydrazines/chemistry , Nerve Growth Factors/chemistry , Penicillium/chemistry , Phenylacetates/chemistry , Chromatography, High Pressure Liquid , Magnetic Resonance Spectroscopy , Molecular Structure , X-Ray DiffractionABSTRACT
During the course of our screening program for low molecular natural products with their ability to potentiate and/or mimic neurotrophic effect of NGF, a novel fungal metabolite, phenylacetic acid hydrazide derivative NG-061 was isolated from the fermentation broth of Penicillium minioluteum F-4627. NG-061 enhanced and mimicked neurotrophic effect of NGF on neurite outgrowth in a rat pheochromocytoma cell line PC12.
Subject(s)
Hydrazines/pharmacology , Nerve Growth Factors/pharmacology , Neurites/drug effects , Phenylacetates/pharmacology , Animals , Cell Hypoxia/drug effects , Cell Survival/drug effects , Cells, Cultured , Drug Synergism , Fermentation , Hydrazines/isolation & purification , Magnetic Resonance Spectroscopy , Neurons/drug effects , PC12 Cells , Penicillium/classification , Penicillium/metabolism , Phenylacetates/isolation & purification , Rats , Rats, WistarABSTRACT
During our screening of microbial metabolites for effective drugs against tumor cell lines, we discovered a new pyrano[4',3':6,7]naphtho[1,2-b]xanthene derivative, FD-594 from the fermentation broth of Streptomyces sp. TA-0256. FD-594 shows moderate activity against tumor cell lines, comparative to that of adriamycin, as well as antibacterial activity against some Gram-positive bacteria.
Subject(s)
Antibiotics, Antineoplastic/isolation & purification , Pyrans/isolation & purification , Streptomyces/classification , Xanthenes/isolation & purification , Animals , Antibiotics, Antineoplastic/chemistry , Antibiotics, Antineoplastic/pharmacology , Fermentation , HL-60 Cells , Humans , Mice , Pyrans/chemistry , Pyrans/pharmacology , Streptomyces/metabolism , Xanthenes/chemistry , Xanthenes/pharmacologyABSTRACT
The structure of a novel antitumor antibiotics FD-594 (1), produced by Streptomyces sp. TA-0256, was determined to have a glycosylated pyrano[4',3':6,7]naphtho[1,2-b]-xanthene skeleton by means of spectral data. The biosynthetic studies of the chromophore of 1 was also carried out by feeding experiments with [1-13C]-, [2-13C]-, and [1,2-13C2]sodium acetate. The labeling pattern was determined by 13C NMR including 2D INADEQUATE experiments, which allowed us to elucidate that the chromophore of 1 is derived from 14 acetate, followed by the loss of one carbon atom.
Subject(s)
Antibiotics, Antineoplastic/chemistry , Pyrans/chemistry , Streptomyces/metabolism , Xanthenes/chemistry , Antibiotics, Antineoplastic/biosynthesis , Magnetic Resonance Spectroscopy , Molecular ConformationABSTRACT
The Ile-Pro sequence of CA074, potent covalent-type inhibitor, is necessary to exhibit the specificity for cathepsin B, but not for papain. In order to elucidate how its sequence binds to papain and why such binding does not exhibit the specificity for papain at the atomic level, two CA074-related compounds, 1 (N-(L-3-carboxyloxirane-2-carbonyl)-L-isoleucyl-L-proline) and 2 (N-(L-3-carboxyloxirane-2-carbonyl)-L-isoleucyl-diethylamide), were designed and their structure--inhibitory activity relationship was investigated by the X-ray crystal analyses of the complexes with papain. The Ile-Pro moiety of 1 was located at the S2 and S3 subsites consisting of Val-133, Val-157, and Asp-158 and of Tyr-61, Gly-66, and Tyr-67 residues of papain, respectively, which is in contrast with the binding of CA074 to S'n (n = 1 approximately 2) subsites in the complex with cathepsin B. Although 2 in the complex with papain showed the similar binding pattern to 1, its inhibitory activity was about two-fold higher than of 1, suggesting the importance of tight S3-P3 hydrophobic interaction for the activity. The difference of the substrate specificity between papain and cathepsin B has also been discussed based on the X-ray results of the present and cathepsin B-inhibitor complexes.
Subject(s)
Cathepsin B/chemistry , Cysteine Proteinase Inhibitors/pharmacology , Dipeptides/pharmacology , Papain/ultrastructure , Cathepsin B/antagonists & inhibitors , Crystallography, X-Ray , Isoleucine , Models, Molecular , Molecular Structure , Papain/chemistry , Proline , Substrate SpecificityABSTRACT
Stachybotrin C and parvisporin, novel neuritogenic compounds, were isolated from the culture broth of Stachybotrys parvispora F4708. Stachybotrin C induced significant neurite outgrowth in PC12 cells and showed cell survival activity in the primary culture of cerebral cortical neurons. Parvisporin demonstrated only weak neuritogenic activity.
Subject(s)
Benzaldehydes/chemistry , Benzaldehydes/isolation & purification , Benzopyrans/chemistry , Benzopyrans/isolation & purification , Fatty Alcohols/chemistry , Fatty Alcohols/isolation & purification , Indoles/chemistry , Indoles/isolation & purification , Neurons/drug effects , Animals , Benzaldehydes/pharmacology , Benzopyrans/pharmacology , Cell Survival/drug effects , Cells, Cultured , Cerebral Cortex/drug effects , Fatty Alcohols/pharmacology , Fermentation , Indoles/pharmacology , Rats , Rats, Wistar , StachybotrysABSTRACT
The structures of stachybotrin C and parvisporin have been determined by spectroscopic analyses and chemical derivatization. Stachybotrin C contains a unique pyrano-isoindolinone ring system, while parvisporin has a hydroxyl farnesyl phenol structure.
Subject(s)
Benzaldehydes/chemistry , Benzopyrans/chemistry , Farnesol/analogs & derivatives , Fatty Alcohols/chemistry , Indoles/chemistry , Farnesol/chemistry , Molecular Structure , Stereoisomerism , Structure-Activity RelationshipABSTRACT
In the course of our microbial transformation study on erythromycin derivatives, Streptomyces hygroscopicus ATCC 31080, which produces a polyether antibiotic carriomycin, was found to transform erythromycin derivatives to their inactivated derivatives. The structures of inactivated derivatives prepared by enzyme reaction using the cell extract, UDP-glucose (or UDP-galactose) and Mg2+ (or Mn2+) were elucidated on the basis of analysis of thei spectral data to be the compounds glycosylated at C-2' of a desosamine moiety, indicating that the enzyme is a macrolide glycosyl transferase (MGT). The MGT activity of cell extract from S. antibioticus ATCC 11891, a producing organism of oleandomycin, could be distinguished from that of ATCC 31080, based on the ability to glycosylate tylosin. We examined 32 actinomycete strains producing such polyketides as macrolide and polyether antibiotics, and found that 15 strains of Streptomyces have macrolide glycosyl transferase activity. It suggests that the MGTs have been distributed among at least polyketide producing Streptomyces strains.
Subject(s)
Anti-Bacterial Agents/metabolism , Streptomyces/enzymology , Transferases/metabolism , Anti-Bacterial Agents/pharmacology , Erythromycin/metabolism , Glycosylation , Microbial Sensitivity TestsABSTRACT
During our screening program of natural products from fungal metabolites for drugs effective against tumor cell lines, we discovered a new isopatulin derivative, pintulin, from the fermentation broth of Penicillium vulpinum F-4148. Pintulin shows weak activity against tumor cell lines, compared to that of adriamycin.
Subject(s)
Antibiotics, Antineoplastic/isolation & purification , Bridged Bicyclo Compounds, Heterocyclic/isolation & purification , Penicillium/metabolism , Antibiotics, Antineoplastic/chemistry , Antibiotics, Antineoplastic/metabolism , Bridged Bicyclo Compounds, Heterocyclic/chemistry , Bridged Bicyclo Compounds, Heterocyclic/metabolism , HL-60 Cells , Humans , Magnetic Resonance Spectroscopy , Molecular Structure , Penicillium/classificationABSTRACT
Three cytotoxic cardenolides, acovenosigenin A 3-O-alpha-L-ramnopyranoside (1), euonymoside A (2) and euonymusoside A (3), were isolated from the woods of Euonymus alata (Celastraceae). The chemical structure of a new cardenolide, euonymusoside A (3) has been elucidated on the basis of extensive spectral analysis and enzymic hydrolysis to be acovenosigenin A (1 beta, 3 beta, 14 beta-trihydroxy-5 beta-cardenolide) 3-O-beta-D-glucopyranosyl (1-->6)-beta-D-glucopyranosyl(1-->4)-alpha-L-rhamnopyranoside. All three showed potent cytotoxic activity against some neoplastic cell lines.
Subject(s)
Antineoplastic Agents, Phytogenic/isolation & purification , Cardenolides/isolation & purification , Digitoxigenin/analogs & derivatives , Plants, Medicinal/chemistry , Animals , Antineoplastic Agents, Phytogenic/pharmacology , Cardenolides/pharmacology , Digitoxigenin/isolation & purification , Digitoxigenin/pharmacology , Drug Screening Assays, Antitumor , Humans , Magnetic Resonance Spectroscopy , Mice , Tumor Cells, CulturedABSTRACT
During our screening program for natural product drugs effective against multidrug-resistant mammalian cells, we have discovered a new delta lactone FD-211 from the fermantation broth of Myceliophthora lutea TF-0409. FD-211 had a broad spectrum activity against cultured tumor cell lines, including adriamycin-resistant HL-60 cells.
Subject(s)
Antineoplastic Agents/pharmacology , Lactones/pharmacology , Animals , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/isolation & purification , Anti-Bacterial Agents/pharmacology , Antineoplastic Agents/chemistry , Antineoplastic Agents/isolation & purification , Drug Screening Assays, Antitumor , HeLa Cells , Humans , Lactones/chemistry , Lactones/isolation & purification , Leukemia P388/drug therapy , Mice , Microscopy, Electron, Scanning , Mitosporic Fungi/metabolism , Mitosporic Fungi/ultrastructure , Spectrum Analysis , Tumor Cells, CulturedABSTRACT
We have investigated the interface between bone and hydroxyapatite (HA) chronically implanted in man. By light microscopy, HA appeared to bind directly to bone without intervening fibrous tissue. By transmission electron microscopy, two patterns were noted: (1) HA either bound directly to bone; or (2) electron-dense material intervened between HA and bone. The orientation of bone collagen fibres likewise showed two patterns: (1) collagen fibres were oriented parallel to the HA; or (2) the fibres were aligned perpendicularly. We have observed similar binding properties of HA to the jaw bone of humans in vivo and in vitro.
Subject(s)
Ceramics/chemistry , Durapatite/chemistry , Mandible/ultrastructure , Maxilla/ultrastructure , Adult , Alveolar Ridge Augmentation , Collagen/metabolism , Collagen/ultrastructure , Female , Humans , Male , Mandible/metabolism , Maxilla/metabolism , Microscopy, Electron , Middle Aged , Prostheses and ImplantsABSTRACT
During the course of our screening program for natural product drugs effective against multidrug resistant cells by using adriamycin resistant HL-60 cells, we have discovered a new 12 membered macrolide FD-895 in the fermentation broth of Streptomyces hygroscopicus A-9561 isolated from a soil sample collected at Iriomote Island, Okinawa prefecture, Japan. FD-895 showed stronger cytocidal activities against in vitro tumor cell lines than adriamycin. FD-895 had the same IC50 values against parent and adriamycin resistant HL-60 cells.
Subject(s)
Anti-Bacterial Agents/isolation & purification , Macrolides , Anti-Bacterial Agents/biosynthesis , Anti-Bacterial Agents/chemistry , Antibiotics, Antineoplastic/biosynthesis , Antibiotics, Antineoplastic/chemistry , Antibiotics, Antineoplastic/isolation & purification , Drug Screening Assays, Antitumor , Fermentation , HeLa Cells , Humans , Magnetic Resonance Spectroscopy , Mass Spectrometry , Microscopy, Electron, Scanning , Molecular Structure , Streptomyces/metabolism , Streptomyces/ultrastructure , Tumor Cells, CulturedABSTRACT
New 18-membered macrolides FD-891 and FD-892 were discovered from the fermentation broth of Streptomyces graminofaciens A-8890 isolated from a soil sample collected at Yamanashi prefecture Japan. They induce morphological changes of HL-60 cells at low concentration below IC50s and have cytocidal activity against in vitro tumor cell lines. FD-891 showed 2 approximately 7 times stronger activity than doxorubicin whereas FD-892 was 20 approximately 100 fold weaker than FD-891 and doxorubicin.