Sesamin is a potent and specific inhibitor of Δ5 desaturase in polyunsaturated fatty acid biosynthesis

Incubation with sesame oil increases the mycelial dihomo-γ-linolenic acid content of an arachidonic acid-producing fungus,Mortierella alpina, but decreases its arachidonic acid content [Shimizu, S., K. Akimoto, H. Kawashima, Y. Shinmen and H. Yamada (1989)J. Am. Oil Chem. Soc. 66, 237–241]. The factor causing these effects was isolated and identified to be (+)-sesamin. The results obtained in experiments with both a cell-free extract of the fungus and with rat liver microsomes demonstrated that (+)-sesamin specifically inhibits Δ5 desaturase at low concentrations, but does not inhibit Δ6, Δ9 and Δ12 desaturases. Kinetic analysis showed that (+)-sesamin is a noncompetitive inhibitor (K_i for rat liver Δ5 desaturase, 155 μM). (+)-Sesamolin, (+)-sesaminol and (+)-episesamin, also inhibited only Δ5 desaturases of the fungus and liver. These results demonstrate that (+)-sesamin and related lignan compounds present in sesame seeds or its oil are specific inhibitors of Δ5 desaturase in polyunsaturated fatty acid biosynthesis in both microorganisms and animals. On leave from Suntory Ltd.     

Regulation of cell growth-dependent expression of mammalian CDC6 gene by the cell cycle transcription factor E2F

CDC6 of Saccharomyces cerevisiae regulates the DNA replication initiation through the origin recognition complex (ORC). Identification of a human homolog of the CDC6 gene (HsCdc6) suggests a universal role of the gene product in DNA replication. Expression of HsCdc6 is growth-regulated. We investigated the molecular basis of growth-regulated expression of mammalian Cdc6. The promoter activity of isolated HsCdc6 upstream region was activated at late G1 and G1/S boundary in the cell cycle of rat embryonic fibroblast REF52 cells by the addition of serum. The isolated promoter was activated by exogenous expression of E2F without serum stimulation. However a mutant promoter lacking the E2F recognition sites failed to respond to serum stimulation and exogenous expression of E2F. Expression of endogenous Cdc6 was induced by exogenous expression of E2F. Therefore, we concluded that the growth-regulated expression of mammalian Cdc6 was mediated by E2F. Moreover, we demonstrated that exogenous overexpression of either HsCdc6 or HsOrc1 failed to induce DNA synthesis unlike overexpression of E2F1, even though E2F1 induced both Cdc6 and Orc1, suggesting that E2F may regulate the expression of another gene(s), besides Cdc6 and Orc1, required for induction of cellular DNA synthesis in mammalian cells.     

Charged amino acids at the carboxyl-terminal portions determine the intracellular locations of two isoforms of cytochrome b5

Outer mitochondrial membrane cytochrome b5 (OMb), which is an isoform of cytochrome b5 (cyt b5) in the endoplasmic reticulum, is a typical tail-anchored protein of the outer mitochondrial membrane. We cloned cDNA containing the complete amino acid sequence of OMb and found that the protein has no typical structural feature common to the mitochondrial targeting signal at the amino terminus. To identify the region responsible for the mitochondrial targeting of OMb, various mutated proteins were expressed in cultured mammalian cells, and the subcellular localization of the expressed proteins was analyzed. The deletion of more than 11 amino acid residues from the carboxyl-terminal end of OMb abolished the targeting of the protein to the mitochondria. When the carboxyl-terminal 10 amino acids of OMb were fused to the cyt b5 that was previously deleted in the corresponding 10 residues, the fused protein localized in the mitochondria, thereby indicating that the carboxyl-terminal 10 amino acid residues of OMb have sufficient information to transport OMb to the mitochondria. The replacement of either of the two positively charged residues within the carboxyl-terminal 10 amino acids by alanine resulted in the transport of the mutant proteins to the endoplasmic reticulum. The mutant cyt b5, in which the acidic amino acid in its carboxyl-terminal end was replaced by basic amino acid, could be transported to the mitochondria. It would thus seem that charged amino acids in the carboxyl-terminal portion of these proteins determine their locations in the cell.     

Approach and structure of stabilizing control system preventing an extension of synchronism loss

Recently, large-capacity power stations have been built far from load centers with long-distance transmission lines. Therefore, if a fault occurs at the line, generators in the large-capacity power station may lose synchronism from the remaining generators [1, 3]. To prevent an extension of loss of synchronism to other generators, fast shedding of some generators is an effective measure. The following methods have been proposed so far: A method [3] based on energy function approach; and a method [2] which estimates and predicts the relative swing among generators using data measured on-line at each generator and proposes shedding some generators for stabilization. This method requires a large communication network for an exchange of data between power stations. This paper presents a new stabilizing method and a newly developed stabilizing control system which does not require a large communication network because it utilizes generator output, voltage and current measured on-line in the vicinity of the large-capacity power station. Using these measured data, the system estimates and predicts the relative swing between generators in the large-capacity power station and the remaining generators. Finally, the number of generators to be shed for stabilization is decided and shedding is performed. Configuration and characteristics of the developed stabilizing control system are shown. The validity of this method is confirmed by simulation and testing using an artificial power system.     

Subtype-specific differences in subcellular localization and chlorethylclonidine inactivation of alpha1-adrenoceptors

Chlorethylclonidine (CEC) inactivation has been used as one criterion to subclassify the alpha1-adrenoceptors (AR); however, the extent of CEC inactivation can vary depending on the CEC treatment. By constructing the FLAG-tagged (N-terminus) and green fluorescent protein (GFP)-fused (C-terminus) alpha1-ARs, we have determined the relationship between CEC sensitivity and the cellular localization of alpha1-AR subtypes using COS-7 cells. In GFP-expressing cells, flow cytometry analysis with anti-FLAG N-terminus antibody detected strong fluorescent signals in most of alpha1B-AR-expressing cells, but low signals in alpha1A-AR-expressing cells. Further examination with confocal microscopy showed that fluorescent signals densely localized intra-cellularly in alpha1A-AR-expressing cells, while most of alpha1B-AR localized on the cell surface. Furthermore, radioligand binding studies with [125I]HEAT showed that CEC (10 microM) treatment of intact cells inactivated approximately 30-40% of alpha1A-AR and >90% of alpha1B-AR, while the CEC treatment of membrane preparations resulted in >80% decrease in the alpha1A-AR density and >90% of alpha1B-AR density, respectively. The results showed that the hydrophilic alkylating agent CEC inactivated only alpha1-AR on the cell surface irrespective of its subtype, and that the subtype-specific sorting is a major determinant for CEC inactivation of alpha1-AR. Subtype-specific cellular localization suggests a new class of functional properties that may explain the signal and functional diversity of homologous alpha1-AR (as well as other G protein-coupled receptors) subtypes.     

Friction and wear of carbon steel near T1-transition under dry sliding

The wear phenomenon of metals under dry sliding is, generally, divided into two modes of severe and mild wear. A discontinuous transition between the wear modes often takes place in a certain load range. The T₁-transition is usually observed at lower levels of load or sliding velocity. There is a great difference in wear rate between severe and mild wear. This indicates that the occurrence of severe wear should be avoided, especially in the field of machine design to prevent energy loss, occurrence of noise and vibration, and life reduction of machines and their components. Therefore, it is important for machine designers to know the relationship between friction and wear and the difference in properties of the wear surfaces in the two wear modes. In this study, wear tests of 0.35% C steel in contact with itself under constant load were conducted in moist air at various contact loads under dry sliding. The friction and wear were measured continuously throughout each test. After the tests, the relationship between friction and wear and the difference in properties of the wear surfaces were investigated in each wear mode. From the results, the upper and lower critical loads (P_acr and P_Acr) appeared between severe and mild wear. The phenomenon of zero wear has been newly found in the early period at very low loads. The zero wear continued for a long sliding distance and then changes to mild wear. The critical load between zero wear and mild wear is defined as P_zerowear. The load was changed once in a step-wise manner from low to high levels in process of test. Since the rubbing history under mild wear condition at the low load in the first stage affected the properties of wear surface, the wear mode at the high load in the second stage changed from ‘mild wear’ to ‘quasi-mild wear’ having a low rate. From the relationship between sliding distance necessary for the appearance of quasi-mild wear and contact load in the first stage, the boundary curve between severe wear and quasi-mild wear in the second stage is hyperbolic. This curve gradually approaches P_zerowear with decreasing contact load. Thus, P_zerowear is one of the important critical loads for elucidating the test results under varying load.     

Phylogenetic analysis of 8 dermatophyte species using chitin synthase 1 gene sequences

Nucleotide sequences of chitin synthase 1 (CHS1) gene of eight species of dermatophytes, Arthroderma benhamiae, A. fulvum, A. grubyi, A. gypseum, A. incurvatum, A. otae, A. simii and A. vanbreuseghemii were obtained and analysed for their phylogenetic relationship. A 600-bp genomic DNA fragment of the CHS1 gene was amplified from these dermatophytes by polymerase chain reaction (PCR) and sequenced. The CHS1 nucleotide sequences of these eight dermatophyte species showed more than 85% similarity between the species. The phylogenetic analysis of their sequences revealed three clusters, the first cluster consisting of A. benhamiae, A. simii and A. vanbreuseghemii, the second cluster consisting of A. fulvum, A. gypseum and A. incurvatum, and the third cluster consisting of A. grubyi and A. otae. The phylogenetic analysis of CHS1 gene in this study will provide useful information for classification and understanding the evolution of these dermatophyte species.     

The tensile properties of an Fe-4 pct Mo-0.2 pct C martensite tempered under applied stress

In Fe-4 pct Mo-0.2 pct C martensite which is a typical secondary hardening steel, premature failure o°Curred in tensile test at 600 °C to 700°C where solute atoms could diffuse easily. To clarify this phenomenon, the quenched specimens were tempered under applied stress and tensile-tested at room temperature. The following results were obtained: (1) Typical intergranular fracture was observed in specimens tempered in a temperature range of 600 °C to 650 °C with tempering times of five minutes to 10 minutes and applied stress (70 MPa to 140 MPa). (2) Based on Auger analysis, this phenomenon was considered to be caused by segregation of P, S, and Mo on prior austenite grain boundaries due to applied stress. (3) The direction of applied stress was found to be very significant. Namely, when the tensile direction was parallel to the applied stress during tempering, the specimen was more brittle, and when tensile direction was normal to the applied stress, the specimen was not so brittle. (4) To reduce this embrittlement, solution treatment temperature was adjusted, and it was found that the embrittlement was considerably reduced both in specimens with fine prior austenite grains and with some ferrite phase on prior austenite grain boundaries. TAKATOSHI OGAWA, formerly with Kyoto University. YOSHIFUMI OHMURA, formerly with Kyoto University. This paper is based on a presentation made at the “pcter G. Winchell Symposium on Tempering of Steel” held at the Louisville Meeting of The Metallurgical Society of AIME, October 12-13, 1981, under the sponsorship of the TMS-AIME Ferrous Metallurgy and Heat Treatment Committees.     

Lateral growth of GaAs on patterned {-1-1-1}B substrates for the fabrication of nano wires using metalorganic molecular beam epitaxy

We have grown GaAs nano wire structures (45 × 20 nm²) buried in AIAs layers by lateral metalorganic molecular beam epitaxy on the terraced sidewalls of mesa-grooved (-1-1-1)B substrates. The growth of GaAs occurred primarily on the sidewall of the mesa-grooves and not on the (-1-1-1)B surface for arsenic pressures greater than 2.0 × 10⁻³ Pa at a substrate temperature of 480°C. An (0-1-1) facet formation during the lateral epitaxy at the intersection region between the bottom (-1-1-1)B surface and the (1-2-2)A sidewall has been directly observed by real-time scanning microprobe reflection high-energy electron diffraction. The growth rate on the (0111) facet was estimated from the variation of its width with growth time.